The Journal of Plastination

Abstracts Presented at The 12th International Conference on Plastination, Murcia, Spain July 11-16, 2004

 

 

Published in the J. Int. Soc. Plast.19: 5-11 and 43-63 (2004)

 

Oral presentations - invited speakers

Preservation and  plastination.  Weiglein AH. Institute of Anatomy, Medical  University Graz, Graz, Austria , Europe.

Background: After death, biological tissues decompose by autolysis or putrefaction . Since the beginning of human life, efforts have been made to stop decay and to keep the body intact, primarily to keep the mortal frame for coming back to life sometime later; later on, interest in morphology made it necessary to preserve human tissue in order to investigate its anatomy. The most important step in preservation was the introduction of formalin by Blum in 1896. Formalin consolidates tissue and quickly stops the decomposition processes . Disadvantages to formalin fixation include unnatural hardening and discoloration of tissues. The introduction of formalin was followed by the color­ preserving embalming solutions by Kaiserling (1900) and Jores (1930). In 1992, Thiel published an article on a new method of color preservation that preserved the human body in lifelike condition (color and flexibility). In addition to the development of embalming solutions that allow preservation for dissection and particularly for surgical training and other invasive procedures, methods were developed for demonstration of human anatomy in museum specimens. In museum specimens preservation of flexibility is not essential. Paraffin impregnation was performed by Hochstetter in 1925. Embedding of organic tissue in plastic was introduced in the 1960's. In 1978 Gunther von Hagens invented plastination. Materials and methods: This technique utilizes both impregnation and embedding, transforming the tissues into plastic with the respective mechanical properties of the polymer. Thus, plastinated specimens are more or less inflexible. In this process, water and lipids in biological tissues are replaced by curable polymers (silicone, epoxy or polyester). These polymers are subsequently hardened , resulting in dry, odorless and durable specimens. Results: Silicone is used for whole specimens (organs, body parts and whole bodies) and thick body and organ slices to obtain a natural look. Epoxy resins are used for thin, transparent body and organ slices excellent for comparison to scans produced by medical imaging techniques. Polyester is used for brain slices to gain an excellent distinction of gray and white matter . Conclusion: Plastinated specimens are perfect for teaching because they show the real specimen, they are easy to handle, almost everlasting and need a minimum of maintenance.

History   of   plastination.   Weiglein  AH.  Institute   of Anatomy,  Medical   University  Graz,  Graz,  Austria, Europe.

Twenty six years ago the first publication on a process for "Impregnation of large specimens  with  polymers" was issued in a German journal (Verhandlungen der Anatomischen Gesellschaft). A year later, in 1979 an article on "Impregnation  of  soft  biological  specimens by thermosetting  resins and elastomers" was published in the Anatomical Record and later on in 1979 the term "plastination" appeared for  the  first  time  in "Emulsifying resins for plastination" (The Preparator) . The first conference on Plastination was held in San Antonio, Texas in  1982. Since  then  plastinators  from all over the world meet every even year for an international conference. Interim meetings, which are primarily workshop based and usually take place in the USA, were started in 1989 in Knoxville, Tennessee. In 1987 the first issue of "The Journal of the International Society for Plastination" was published . In  1996  the first issue of the Current Plastination Index - an index listing all publications dealing with plastination - was issued and updated in 2000. This CPI 2000 lists almost 800 publications dealing with plastination  in more than 80 different journals. However, it was not before 1996 that the International Society for Plastination (ISP) was officially launched during the meeting in Graz. Since then, the ISP serves as a forum for the exchange of information about  Plastination,  which  is accomplished by the publication of the journal , by  holding conferences, meetings and workshops on a regular basis and by our web page and list server.

Principals of plastination. Henry RW. Department of Comparative Medicine, College of Veterinary Medicine, University a/ Tennessee, Knoxville, TN, USA.

Plastination, the exchange  of tissue fluid for a curable polymer, is an intriguing method for preserving biological tissue in a cost effective manner. Plastination has four basic steps: Specimen  preparation , Dehydration, Impregnation and Polymerization. To get the best specimen for your effort, plan the final product before production commences. Have a road map. The raw materials (specimen) likely should undergo some major renovations during  the  specimen  preparation stage. Do not attempt to show every feature on one specimen. Once the specimen is prepared or during preparation it may  be fixed in  2 - 20% formaldehyde solution. After fixation for a desired period, the fixative is flushed out to prevent contamination of the acetone and hence the distillation unit with paraformaldehyde. Dehydration   is  best   carried   out  with   cold   (-25°C) acetone - "Freeze substitution" . Consistently , the best specimens will result from this type of cold dehydration . Cold room or room temperature acetone may be used with good results. For large specimens a higher percentage graded acetone series may be utilized with success. Your favorite graded series of alcohol may also be used for dehydration. However , excessive shrinkage occurs and a suitable intermediary solvent will need to be used for the polymer exchange, since alcohol is not an appropriate solvent for this exchange. The exchange of the solvent for the polymer is called "forced impregnation". Impregnation takes place in a vacuum chamber and will occur if the solvent boiling point is low enough or vapor pressure is high enough such that it will sufficiently and steadily vaporize as the pressure is lowered in the plastination kettle. As well, the polymer must have a boiling point that is high enough that the polymer will not vaporize. Acetone and methylene chloride (dichloromethane) (MeCl) both are suitable solvents for the impregnation process. However, MeCl is not a dehydrant. Therefore it can only be used after the specimen has been dehydrated. Silicone, epoxy and polyester are the commonly used polymers. Silicone is generally used to plastinate whole or portions of the body or organs. It may also be used for preserving sections from various regions of the body. Both epoxy and polyester are used primarily to produce thin sections (2-6 mm) of the body. Classically, the main use of polyester has been for head and brain slices. Special processes for both epoxy and polyester polymers  maybe  used  for  producing  thin  sections, <1mm. Rendering the polymer ready for polymerization is accomplished by adding a catalyst. In the case  of epoxy and silicone, the catalyst is recommended to be mixed with the polymer prior to impregnation. While polyester's catalyst is UV light and a catalyst does not have to be mixed in the impregnation polymer. The reactivity of the catalyzed polymer is retarded by cold temperature. Epoxy and polyester both yield a hard sheet. The plastinated slice is placed  between  two plates of glass separated by a gasket and the remaining space is filled with polymer. Silicone, on the  other hand, produces a specimen with some degree of flexibility . Flexibility of silicone specimens seems to be dependent on the thickness of the tissue and possibly tissue density. Flexibility may be enhanced by the addition of a chain extender which may be added to the impregnation reaction-mixture or to the specimen after impregnation. Plastination polymers are not inherently flexible and durable after curing when compared  to RTV silicone. RTV contains fillers that provide  its great   flexibility.   However,  any   of   the   plastination silicone polymers yield similar finished products if the ingredients are added in a similar  sequence.  The standard Biodur method of combining polymer and catalyst in the impregnation mixture consistently yields excellent specimens. Polymers  used  today  for plastination are routinely used in the polymer industry . The biggest difference in silicone polymers is their viscosity which ranges form 40 to 1,000 estk. Polymerization (hardening or curing) of the polymer is promoted by adding a hardening agent that promotes cross-linking of the silicone chains. The Dow/Corcoran method adds the cross-linker and catalyst in the reverse of the classic method and specimens generally have poorer surface detail. Thorough mixing of ingredients is a necessity. Patience and accuracy will yield  a durable specimen.

Dissections before, during  and  after  plastination. Boyes R, V Kippers. School of Biomedical Sciences and Plastination Laboratory, Faculties of Biological & Chemical Sciences and Health Sciences, University of Queensland, Brisbane, Australia.

Novice plastinators may assume that dissection of a specimen must be completed prior to commencement of the plastination process. However, further dissection is possible both during and after plastination, which often results in enhancement of features  chosen for display. The purpose of this presentation is to generate discussion about dissection techniques of plastinated material , seeking the input of plastinators  who can share their experiences and provide tips for their colleagues. The presentation will briefly discuss pre­ dissection techniques for the benefit of  novice technicians. The importance of dissection prior to plastination is paramount  because the final  appearance of the specimen is dependent on the time, effort and technical expertise of the dissector. All extraneous connective tissue must  be  carefully  removed ; underwater dissection magnifies the connective tissue which can then be carefully removed to produce a smoother surface with muscle  fascicles  enhanced. Stents can be used to maintain the shape and size of the lumen of vessels and orifices. Other devices are used to maintain correct  relationships  between  structures during the process. Some of these devices can be retained while others are removed during  the plastination process. Use of compressed air during the plastination process produces organs with  a  realistic size and shape, such as the lungs,  stomach  and intestines. Sections of organs can be produced after plastination of complete organs. As an example, brain slices   can   be   produced   from   whole   brains,  often producing slices of better quality than the alternative of slicing the brain prior to plastination. In terms of presentation of plastinated specimens, there is definite benefit in using a range of techniques to highlight specified features and to maintain the  correct appearance of structures that tend to collapse during the plastination process.

Principles  of  silicone  plastination  techniques.  de Jong KH. Academic Medical Center. Anatomy and Embryology. Meibergdreef 15, 1105AZ, Amsterdam, N H., The Netherlands , Europe.

Since 1979 different techniques of plastination are  in use in many institutes all over the world. These techniques can be divided in two groups: sheet plastination using epoxy or polyester and silicone plastination. Although the basic principles  are  similar for both procedures, differences exist for each polymer used and the flexibility of the resulting specimen. Most people starting with plastination will begin their career as a plastinator using the silicone technique due to factors as: easy to learn, relatively low  starting costs, easy obtainable equipment, etc. Therefore  this technique will be explained and discussed in this presentation and special attention will be paid to tips, tricks, and pitfalls in each step of the procedure . The main steps in silicone plastination are: fixation, dissection, dehydration, (forced) impregnation, and curing. 1. Fixation: this step is not necessary. Both formalin or otherwise fixed specimen or fresh specimen can be plastinated. 2. Dissection: special care must be given to dissection, only the best  dissected  specimen will yield a nice looking plastinated specimen. 3. Dehydration: is performed  by  submerging  the specimen in subsequent acetone baths (100%) until the remaining amount of tissue water  is <l%. Dependent on the desired degreasing effect of the acetone, the specimen can be left in the last acetone bath longer than desired for the dehydration . 4. (Forced) impregnation: is performed by submerging the specimen in a bath of monomer silicone with an increasing vacuum, thus boiling the acetone out of the specimen and dragging the silicone into the specimen. The rate of impregnation is monitored by watching the amount and size of the escaping acetone bubbles. 5. Curing: is a two-step procedure . First elongation of the monomer silicone molecules, caused by a "chain elongator" and second cross-linking of the elongated silicone  molecules, caused by the "cross-linker". The latter step forms a 3- D spatial meshwork of flexible molecules  throughout the specimen, thus producing a lifelike, dry, odorless and easy to handle anatomical specimen.

Silicone   plastination,   room    temperature methodology: Basic tech niques, applications  and benefits for the interested user. Glover R. Director, Plastination Laboratory, University of Michigan, Ann Arbor , MI, USA.

In July 1998, Dow Corning introduced a room temperature preservation process to those attending the Ninth International Conference on Plastination in Trois­ Rivieres Quebec, Canada. This was a significant event in the history of the society because , up until that time, Biodur S10 methodology was the only method available to those interested in  doing tissue impregnation with silicone. Prior to the Quebec meeting, Dow Corning, through its chemical distributor Corcoran Laboratories, selected three active plastination laboratories and provided them with chemicals and a description of their newly patented process . The laboratories were asked to experiment with the process and evaluate it to see if it was effective and had significant technical benefits that might be of interest to the society as a whole. The laboratory at the University of Michigan, Ann Arbor, Michigan, was one of the three  laboratories invited to experiment with the new process. Dow Corning's invitation was accepted . Since, the Biodur S 10 process had been used in the lab for over six years, both processes and chemicals could be compared and evaluated. This presentation  will present  the highlight of six years of experience using exclusively the room temperature process in  our laboratory since its introduction. The focus will be on: the significant benefits inherent in the room temperature process - the things that make it a flexible, time efficient and a cost effective approach to silicone preservation .  We  hope that this information will give conference attendees who are familiar with the room temperature process an opportunity to understand its basic methodology thus enabling them to decide whether or not to adopt this process for use in their own laboratories .

Maximizing anatomical concepts  with  sheet plastination. Cook PR. Department of anatomy with Radiology, University of  Auckland , Auckland New Zealand.

Background: As sophisticated  diagnostic  technology has become commonplace, our teaching program has been tailored to maximize clinical relevancy of the learning experience, with practical laboratory demonstration routinely provided by radiology, ophthalmology, orthopedic, otolaryngology, surgical registrars and an anatomical pathologist. Clinical procedures, pathological observations and diagnostic methods are as integral a part of the education process as is the gross structure of the human body. Plastination has proven to be a vital tool in the enhancement and broadening of anatomical concepts. Materials and Methods : While the University of Auckland utilizes all current Biodur based plastination techniques - S10 silicone, P.E.M. polymerized  emulsion and E20 injection casting - to provide an excellent three dimensional realization of the human body, it is the El2 epoxy method of producing serial-sectioned anatomy that has provided significant educational opportunities. Whole cadavers or regions were cut into 2.5mm thin transverse, coronal or sagittal sections, depending upon the desired result. Sections were dehydrated , degreased, impregnated , cast and cured according to standard epoxy plastination protocols. The completed sections are smooth, semi-transparent , durable and offer an exceptionally clear degree of anatomical detail not seen in traditional cross sections or wet gross specimens. Results: The El2 epoxy method of sheet plastination permits, the finished sections to be utilized for correlation and clearer interpretation of similar magnetic resonance images (MRI) and computer tomography (CT). When used in student microscopy laboratories, El2 anatomical structures may  be magnified considerably to the submacroscopic level allowing an important conjunct  between microscopic anatomy and macroscopic anatomy. Whereas standard histological sections provide detail only within predetermined parameters dictated  by the physical limitations of the dimensions of the actual microscopic slide itself. El2 sections contain a high degree of detail whilst, importantly , retaining the in situ structural integrity of the entire region in a complete and uninterrupted state. Conclusion: The unique properties of the E12 process results in the linking together of three distinct disciplines, namely cross-sectional anatomy, radiology and histology from just the one specimen.

Principles of polyester plastination techniques. Weiglein AH. Anatomical Institute , Medical University Graz, Graz, Austria, Europe.

Background: Plastination of brain slices can be done by the standard silicone technique or by the polyester techniques . The silicone technique is used to produce thick, opaque and natural looking brain slices. The polyester techniques (P 35 or P 40) are used to produce thin semitransparent brain slices. Materials and methods: For polyester plastination the recommended procedure is as follows: Fixation : Brains are fixed the usual way with 10% formaldehyde alone, since other fixatives  may  interfere  with  this  procedure.  Slicing: Brains are sliced with a meat slicer into 4-8 mm thick slices. To avoid disintegration , a piece of wet filter paper trimmed to the size of the brain is placed on top of the brain slice before slicing. The slices are placed on stainless steel grids and the grids are put together into a stainless steel basket and stored in cold fresh tap water. Flushing: The basket of brain slices is rinsed with cold tap water overnight and thereby cooled down to 5°C. Dehydration: The basket of brain slices is submerged in 100% acetone at -20° for three days. Immersion: The basket of brain slices is removed from the acetone and immediately submerged in the polyester-mixture for 1-2 days at 5°C. Forced impregnation : The basket with brain slices, submerged in the polyester-mixture is exposed to vacuum  for 24 hours either at room temperature (not below 10 mm Hg) or at -20°C (not below 2 mm Hg). Glass chambers: The impregnated slices are enclosed in 5-10 mm wide glass chambers. Two glass plates are separated by a silicone gasket and held together by fold-back clamps. The glass chambers are filled with  the polyester mixture. Curing: The glass chambers are exposed to UVA-light for 3 hours (400 Watts at 25 cm distance from the chamber). During this procedure the chambers must be cooled either by ventilators or by blowing compressed air over both sides of the chambers. P 40 slices are cured by UVA light only, whilst P 35 slices need additional heat curing. Following light curing, the P 35 glass chambers are exposed to 45°C for 5 days in a well-ventilated oven. After curing is finished the slices are dismantled, trimmed by a band saw and the edges smoothened by a belt sander. Results : Polyester plastination of brain slices results in 5-10 mm thick, easy to handle plates enclosing the impregnated brain slices. The semitransparent brain slices are particularly superb in gray and white matter distinction. Conclusion: Due to the detailed anatomy shown in polyester brain slices and due to their comparability to CT and MRI scans,  they are excellent for teaching neuroanatomy.

Polyester plastination  techniques:  Specific  troubles and problems. Henry  RW.  Department  of Comparative Medicine, College of  Veterinary Medicine, University of Tennessee, Knoxville, TN, USA.

Polyester plastination has been a popular and convenient method to preserve brain slices for nearly two decades. It involves slicing thin (2-6 mm) slices of well-fixed brain, dehydrating, soaking in polymer, impregnating with polymer and finally placing the slice in a flat chamber with polymer to make the final sheet. In the resulting slice, gray and white matter are clearly differentiated. Two widely used polymers have been used, P35 and P40. P35  was  introduced  first  and yielded brain slices of unparalleled beauty, clarity and definition of white and gray matter. P40 was introduced 10 years later and is a shorter and less cumbersome technique. It also yields a remarkable differentiation of white and gray matter . However, the P40 polymer has a specific problem when used on brain tissue, orange spots in the gray matter and when uniformly distributed throughout the gray matter, it may resemble the coloration of P35 slices. No predictable mechanism has been developed to determine if the spots will or will not develop in brain slices. Some theories of the cause or reason for the orange spots have been offered (tissue peroxidase, fixation or fixatives, incomplete impregnation) but no clear resolution has been offered. An additive has been developed to correct this problem and it is seemingly beneficial to minimize spots. Shrinkage of 4.5 to 7% has been reported depending on whether cold or warm temperature impregnation was used to produce P40 slices. P35 slice production takes ten to twelve days, while P40 slices take only four or five days to complete. P35 slices use twice  as  much glass as P40 slices. Catalyst must be used in the immersion and impregnation baths of P35, therefore more polymer is used and impregnated slices should be cast within a few days. However, P40 slices can be held for several weeks after impregnation since there is the possibility of using no catalyst in the  impregnation bath. Recent results show that P40 can be used successfully to produce slices from all regions of the body. P35 and P40 slices remain fully transparent with no yellowing over time. If the P35 or P40 impregnated slices remain in the polymer for extended periods of time, the slices may adhere to the grid spacers that are used to separate the tissue slices.

Principles of epoxy  plastination  technique  (E12). Sora M-C. Plastination Laboratory, Center for Anatomy and Cell Biology, Medical University of Vienna, Vienna, Austria , Europe.

Background: The E12 plastination process is a well­ established preservation technique used  for demonstration in teaching and  also  in  research . Material and Methods : Materials and slicing: For El2 plastination , fresh tissue which is frozen at -80°C for one week is used . Next slices with an average thickness of 3 to 5mm are  cut. Between the sections, 1 mm of tissue is lost due to the thickness of the saw blade. The slices are stored at -25 °C over night. Dehydration and Degreasing:  Cold  technical  quality  acetone  (-25°C)  is used for dehydration of slices. Each slice is placed between plastic  grids to allow better  circulation of the dehydration fluid. Dehydration time for the slices is 7 days. The acetone is changed after 3 days and its concentration will have dropped to around 96%. After 4 days in new acetone, the final concentration of the acetone dehydration bath should be 99%. When dehydration is complete, the freezer is disconnected and the temperature allowed to increase  to  room temperature (+15°C). Next the acetone is changed for room temperature methylene-chloride (MeCl) for degreasing. Degreasing is finished after 7 days. Impregnation: Impregnation is performed at +5°C using an epoxy reaction-mixture (El2). The slices  are submerged in the E12 reaction-mixture and placed in a vacuum chamber, directly out of the methylene-chloride bath. Pressure is continuously reduced  over  the  next two days down to 2mm Hg. Temperature  is kept under surveillance in order to avoid El2 crystal formation which may take place if temperature decreases below 0°C. Casting and curing: The  slices  are  cast between two sheets of tempered glass and a  flexible gasket is used as a spacer (4mm). The following E12 reaction­ mixture is used for casting: E12/E1/AT30  (95 :26 :5). The slices are placed  between  glass plates, sealed with a gasket  and clamps, and the chambers filled with the casting mixture . After filling, they are placed in a vacuum chamber at 3 mmHg for one hour to remove small air bubbles present in the resin. Large bubbles are removed afterwards manually. After  bubble  removal, the flat chambers are placed nearly horizontal (inclined at 15°) and left until the next day.  The  polymer becomes more viscous and sticky  and after  one more day the flat chambers containing the slices are placed in an oven at 45°C for 4 days. After removal  from  the oven, the flat chambers are cooled to room temperature . The glass plates are removed carefully and the epoxy sheets are trimmed as desired. Results :  The transparency and color of the slices are perfect and shrinkage is not evident. The finished E12 slices are semi-transparent , easy to orient and offer a lot of anatomical detail down to the  sub macroscopic  level. The transparent  areolar and adipose tissues contrast perfectly with the muscle tissue and epithelial parenchyma. Conclusion : Since the beginning of plastination , the E12 technique is still the  selected method for producing transparent body  slices. Transparent body or organ slices are used for teaching and research , because they allow study  of  the topography of all body structures  in  a  non-collapsed and non-dislocated  state. In addition, the specimens are useful in advanced training programs in sectional topography (resident training in CT and NMR) .

Tips and tricks in plastination. Weiglein AH. Institute of Anatomy, Medical University  Graz,  Graz, Austria, Europe.

Practically every  step in plastination  can be improved to save time and/ or money or  to  improve  the production of better plastinated specimens. A good specimen can only be the result of a well planned dissection: It is advisable to leave  structures  longer than they should be in the final specimen . E.g. blood vessels, nerves, muscles and tendons can be trimmed after the plastination process, which results in smooth sections. Hollow organs (lung,  heart,  stomach, intestines, large blood vessels)  must be  dilated  during the whole plastination process, at least  during dehydration and curing to keep their form. Several studies have shown that specimen shrinkage is lowest when cold acetone  is  used  for  dehydration. Dehydration at room temperature with acetone or methanol can result in up to 50 % shrinkage (20-25%) compared to dehydration at minus 25°C (10-15%). For improvement of the dehydration process, the basis for dehydration must be understood: It is a dilution process during which  the water  concentration  of the  specimen is minimized to zero by diluting the water inside the specimen with acetone. Basically, a specimen could be dehydrated in one step by submerging it in pure acetone 100 times the volume of the specimen. E.g.: one human brain weighs 1,3 kg which contains  approximately  1.0 L of water. If this brain is submerged in 100 L of pure acetone this would result in a 99% concentration over a period of time. To reduce the amount of acetone needed it is recommended to work with a 1:10 specimen­ acetone ratio. In the same case (1 human brain 1 L water) the first acetone bath of 10 L pure  acetone results in a 90 % concentration, the second in 99 % and the third in 99.9 %. Thus, it is  recommended  to  use three baths of pure acetone each with 10 times the specimen's volume . This procedure is the fastest possible, but also the most expensive one. To save money, acetone of lower concentration (down to 80%) can be used to start dehydration ; the last bath, however, would have to be pure acetone to get complete dehydration. Acetone of lower concentration  (below 75%) can be recycled with commercial or self made solvent recyclers. To minimize shrinkage it is recommended: To use cold (-20 to -25°C) acetone for dehydration, To dehydrate to  99-100%, To impregnate completely (down to 0-5 mm Hg), To use fast curing, and To dilate hollow organs.  High  flexibility can only be reached in thin organs; e.g. in the stomach and the intestines. If a specimen is too dark after plastination it can be brightened up by wrapping it in cloth soaked in 10 % peroxide for several minutes or hours. To keep some flexibility it is recommended: To use no hard fixation (no formalin), To use less S3 (hardener I chain­ extender), To dilate hollow organs, To place plastic foil in between anatomical layers, and To wipe  and  move the specimen during curing process.

Equipment and costs for plastination. Diz A. Departamento de Anatomia y Anatomia Patol6gica Comparadas, Faculty of Veterinary Sciences. University of Cordoba, Cordoba, Espana, Europe.

The following review is suggested for  beginners  and will give you a general  overview  of  necessary equipment and cost of a silicone plastination laboratory using  the  standard  S-10  technique  and  sheet plastination (E-12 and P-35/P-40 techniques). The proportions of the  equipment depend on the kind, size and number of specimens to be plastinated and on the plastination technique being used. Generally it is advisable to start with the standard silicone technique at a low level and to expand the laboratory set-up  after some experience has been gained. All of the various methods of plastination have the common principle: the use of a vacuum chamber and a vacuum pump  to extract the solvent saturated within the specimen. A means of regulating and monitoring the amount of pressure is also vital to the success of the plastination process.

Basic equipment for silicone plastination :

3+ plastic (polyethylene or polypropylene) (PVC or polyester are not  recommended),  aluminum  or stainless steel containers corresponding to size of specimens for fixation, dehydration  and  degreasing, but large enough to  hold  the  specimens  and dehydration agent at a ratio of 1:10.

1+ deep freezers large enough to hold at least 2 of the dehydration containers and 1 vacuum chamber corresponding to size of specimens.

1 Grid basket (made of acetone resistant plastic, aluminum or stainless steel) to contain the specimens for dehydration and forced impregnation .

1 Acetonometer 0-100% and  1 acetonometer  90-100% to monitor dehydration . Ideally no more than 1-2% at the most of water, is allowable to remain in the specimen.

1 Vacuum chamber of stainless steel corresponding to size of specimens with 3 connecting ports and a glass plate on top for a lid and for visual control of the impregnation process.

1 Rotary vane vacuum pump, suitable for solvents, suitable for size of vacuum chamber and vacuum tubing. The pumping speed needed  in plastination depends on volume of impregnation bath. Usually pumping speed is lm3/h per 10 liters of impregnation bath. If in doubt, a lower pumping speed is advisable .

1 Separator for oil and solvent in the pump exhaust (not indispensable): a condensation and  collecting  vessel for extracted solvent and oil vapor given off during forced impregnation . Placing it in the deep freezer allows to recover almost  100% of  extracted  solvent. At room temperature recondensation is only partial.

1+ vacuum  adjustment valves:  needle valve for manual control of forced impregnation via admission of external air. System of two parallel-grouped valves allows finer adjustment of vacuum than one valve .

1 Manometer to monitor vacuum during the first two thirds of forced impregnation.

1 Bennert-Manometer to monitor vacuum  during  the last third of forced impregnation.

Vacuum tubing to connect vacuum chamber to: vacuum control and adjustment devices, to vacuum pump and to separator for oil and solvents.

1 Gas curing unit (for curing silicone impregnated specimens) corresponding to size and number of specimens, consisting of a plastic, aluminum or stainless steel container  provided with  sealable lid,  1 or more closeable  glasses for gas curing  and desiccator agent, 1 small membrane pump to vaporize silicone hardener (S6) and 1 draining grid. Moreover, an air compressor is recommended to dilate hollow organs and a distillation unit for recycling acetone.

For sheet plastination ,  all the equipment described above is necessary except the gas curing unit; moreover, the following equipment should be available : 1 Rotary meat slicer for organ slices

1 Band saw with coolable guide stop for frozen specimens and body specimens containing bony structures.

1 Thickness gauged.

Sheets of polymer gauze and polymer or stainless steel grids and nets for stacking and stabilizing of body and organ slices during  dehydration,  degreasing  and forced impregnation.

Equipment for flat chambers : Toughened glass plates (3-4 mm. thick, free of scratches), silicone gaskets as spacer for flat chamber (thickness depending on thickness of slice), foldback clamps for assembly of flat chambers, 1 or more flat funnels to fill flat chambers with resin, wire in order to positioning the slices.

Oven (50°C) for curing of specimen slices according to E-12 and P-35 technique (for P-40 technique it is not necessary) .

1 U.V.A. light curing unit for curing of plastinated slices with P-35 or P-40 Techniques, constructed with two standard fluorescent light fittings equipped with fluorescent U.V.A. tubes. It must integrate a fan for cooling and time switch.

In a plastination lab, remember that  acetone fumes  are an explosion danger, so all precautions be taken to minimize the risk of explosion. Great care must  be taken to isolate the electrical elements in the lab. A general hint in plastination is that if a household deep­ freezer is used, the interior lighting has to be removed. Moreover, the compressor and  motor  should  be removed and re-situated outside the freezer in the adjoining room for maximum  hazard  control.  In  case the compressor and motor are not dismounted,  the freezer should  be disconnected from power  supply before handling of acetone inside the  freezer.  The vacuum pump must be sited outside the lab for the same reason, and an acetone fume detector  should  be installed in the plastination lab. In order to reduce costs in plastination, much of the equipment required for the setting up of a small to medium sized plastination lab is often readily available in most universities and other institutions, so major capital expense can be negligible. It is desirable to acquire most of the equipment  from local suppliers and to recycle  used  acetone,  polymers and other chemicals  in the lab. Finally , a great deal of talent and imaginative flair can often produce  good quality plastinated  specimens with limited means.

Plastination: A tool for education: Lozano// S. Department of Biomedical Sciences, University of Hawai'i, School of Medicine, Honolulu, HI, and SURFdriver Software, Kailua, HI, USA.

Background : Problem based  learning  (PBL) pedagogical strategies have become more prevalent in medical school curricula . PBL  focuses on small group interaction and experiential, case-based learning. The clinical problem is the vehicle for learning and it is peer-taught and  tutor-mediated.  A  specific  case scenario is developed and then the small group works through the case to establish  a differential  diagnosis in a logical, hypothesis driven process. PBL also provides the opportunity to introduce  virtual  reality  (VR) teaching tools into the curriculum that includes anatomically relevant  clinical  conditions.  The  purpose of this paper is to review the efficacy of VR presentations using anatomical  animations  generated from plastinated tissues. Materials and  Methods:  A PBL case involving  an epidural  hematoma patient  was developed. An adult human brain was obtained, dehydrated, impregnated (P40), cast and serially sectioned. Cross sections were obtained and a base illustration was prepared to represent an epidural and subdural hematoma. Interpolated frames were established based on a linear transformation between 28 key frames emphasizing compression of the parietal region and cranial nerve III ipsilaterally . The animation was presented within the context of a child  who fell sustaining a head injury. It was posted on a website and a questionnaire was developed and circulated  to medical students participating in the PBL case. Results : The evaluation showed that the animation aided the students' understanding of the health care problem (average score of 1.36; SD = 0.59; n = 19, on a scale of 1-5 with 1 being "Strongly Agree"). Numerous additional comments were presented indicating that the epidural hematoma provided a useful PBL resource . Conclusion : Plastination provides a powerful technique to develop supplemental learning tools for VR delivery of PBL concepts. Supported, in part, by OAT HHS 2 DlB TM 00003-02.

Plastination, a useful  tool  in  teaching  clinical anatomy. Riederer BM E  Musumeci,  B  Duvoisin 2 FJW Lang1 . Department de Biologie Cellulaire et de Morphologie , University of Lausanne, Service 1d'OLR et 2Radiologie , Lausanne, Switzerland , Europe.

Plastination  is essential  in teaching  several  courses  at the  University  hospital  at  Lausanne. Previous  reports outlined the necessity to reduce the number of courses and dissection hours in teaching human gross anatomy in the  2nd year  of medical  studies  at the  DPCM.  In addition,  a  report on another  module, where plastination  is advantageous  for preservation  due to a repetitive  use of specimens.  One module, such  as the lower limb, usually consisted of 27 hours of dissection , which  was  distributed  over  9  afternoons,  but  for the novel, the course had to be reduced to three blocks of two hours of self-directed learning.  Assistants predissected   specimens   several   months   in   advance which  familiarized  them  with  the  teaching  material , since they were to supervise and guide the students and furthermore  gave  them  a  good  opportunity  to  learn topographical   anatomy  in  more  detail.  This  allowed ample time for plastination  of the dissected specimens by the standard  S 10 procedure.  Further  details on the technique, references and links can be obtained on our web  site: (http://www­.ibcm.unil.ch/teaching/anatomie/plast/indexplast.html)  . During the two first hours of the self-directed teaching module, cutaneous  innervation  and  articulations  were studied; followed  by  the  topography  of the  thigh  and popliteal region, and finally lower leg and foot were studied. The plastinated specimens supplemented the teaching material, which consisted in part of wet specimens, usually kept in 50% alcohol solution between courses. Unfortunately , delicate  nerve structures of different cutaneous regions  resisted  only for a short time the inquisitive  approach  of  our students. Even nerves sutured to the tissue were soon tom apart. Therefore, the use of plastination resulted in more resistant specimens. This module will now be adapted for 1st year medical students which is a challenge because of the large number of student (over 400 students). The plan is to include plastinated specimens in this new module, because of the previous good experience. Furthermore ,  the  plastinated specimens were also very welcome by the students, especially for individual study before exams. For a second teaching course for 5th year medical students on the maxillary sinuses, samples were prepared and plastinated . Each year, human heads obtained from the local donation program had to be prepared again, therefore, these permanent preparations are  very welcome . Six heads were prepared according to the traditional protocol, in addition red silicone color with radio-opaque material (Lipiodol) was injected into the internal carotid artery . Several surgical interventions (unciformectomy , frontomeatotomy, ethmoidectomy or antrostomy) were applied to some blocks, furthermore some arteries were prepared prior to plastination . The maxillary blocks were subjected to CT scans before and after plastination to determine shrinkage effects due to the plastination process. Bony structures, although fragile, were far less prone to shrinkage than sinus mucosa. To illustrate the trajectories of several arteries and to illustrate surgical interventions an endoscopic approach was chosen. However, the endoscopic examination proved sometimes more difficult due to the increased rigidity of plastinated specimens. In conclusion, advantages of introducing dissected and plastinated tissues are manifold. Less bodies are used because of a reduced necessity to prepare new specimens every year. Furthermore , delicate structures become more resistant and well dissected tissues is preserved for a long time. In addition, plastinated samples can be used also  outside  the  dissection  hall. The introduction of plastinated samples  adds  another tool to teaching clinical and applied gross anatomy.

Plastination: A tool  for  research:  Zhang  M. Department of Anatomy and Structural Biology, University of Otago, Dunedin, New Zealand.

Background:  Since  its  innovation  by  von  Hagens  in 1979, the plastination technique has been widely and extensively used in teaching. There are also increasing number of researchers who take advantage of the sheet plastination technique for 3-D reconstruction  and CT/MRI correlation . However, the application of plastination for other research is not very common. For instance, the PubMed search for "Sheet plastination" reveals that up to April, 2004, there are 80 publications , over 70% of which were related to teaching, 3-D reconstruction and technical  development.  Even  less hits (5-6 papers)  were  generated  for  "Sheet plastination". Moreover, the areas covered by these published studies were relatively  limited.  Based  on those published studies and our own experience, we believe that the  potential  of  the  plastination , particularly El2 sheet plastination , as a tool for morphological research should be further and widely explored. Materials and Methods : A literature review. Results: It was found that sheet plastination techniques are extremely useful for the investigation of the spatial relationships between fine structures and the macro configuration of connective tissue structures, e.g. tendons, ligaments, fascia,  sheathes  and  septa.  With pre- or post-treatment , the sheet plastination technique can be modified to meet specific objectives, such as localization of micro-venous valves and  collagen components, as the prepared specimen can be examined under optical, electron and confocal  microscopes .  On the other hand, however, there are some limitations, for instance, it is time-consuming and expensive. These limitation may limit the size of samples and prolong the study. The tissue loss due to slicing may also obliterate vital information skewing results of those studies on an unevenly distributed structure. Conclusion : The plastination  technique, particularly  sheet plastination , is a unique tool for the morphological investigation at the macro- and microscopic level.

Plastinated specimens in the minimally invasive surgery center. Sanchez Margallo FM. Minimally invasive surgery Center, Caceres, Espana, Europe.

Introduction : Now days, surgery is a multidisciplinary specialty that involves different technological fields. Surgeons should be able to  perfectly  understand  and use the latest technological advances. The Minimally Invasive Surgery Centre (MISC) has always promoted totally practical surgical training programs , where experimental models such as plastinated organs play an essential role, especially in the basic steps of the surgeon's learning curve. The advantages of minimally invasive surgery over conventional approaches are already proven . One of the main problems of minimally invasive procedures is that their clinical application must be preceded by a highly specialized training program. This protocol should be divided in different levels of progressive difficulty , where experimental models should be used on the basic steps. Over the last years, plastination has proved its usefulness in the fields of anatomy, pathology, surgery and radiology. This report is intended to discuss the advantages of the use of plastinated organs as adjunct tools for research and teaching in minimally invasive surgical techniques. Advances in plastination over the last years have allowed the development of new research areas in the fields of anatomy and on several diseases. Simulation of clinical situations would allow clinicians to profit from practical training courses. From our point of view, the development and clinical validation of new training protocols that include the use of plastinated organs is of the greatest importance. This would decrease time and costs of the learning curve while minimizing risks for both patients and clinicians. Materials and Methods: Plastinated models of the canine digestive system were prepared at the Veterinary Anatomy Department of the University of Murcia. These organs were used as experimental models on the Theoretical and Practical International Course on Digestive Endoscopy, held at the MISC. Attendees of the course were trained on both basic diagnostic  and therapeutic endoscopic procedures, such as esophagogastroduodenoscopy, colonoscopy, biopsy, cytology, polypectomy and foreign body retrieval, using plastinated organs as a step prior to the "in-vivo" procedures on experimental animals. Finally, plastinated organs were compared with experimental animals in terms of effectiveness and usefulness for surgical training programs. Results : The course attendees agreed on the usefulness of plastinated organs, especially in the first steps of their learning curve. Endoscopic evaluation of the gastrointestinal mucosa, as well as basic therapeutic techniques were successfully carried out on plastinated models. Intimate knowledge of the canine gastrointestinal anatomy was easily learned in the plastinated organs, which is a basic prerequisite for the performance of "in-vivo" endoscopic explorations. Training on therapeutic endoscopic techniques (cytology, biopsy, polypectomy and foreign body retrieval) using plastinated models provided the attendants with the necessary  basic practice for safe "in-vivo" endoscopic procedures. Conclusions: The use of plastinated organs is a useful tool for surgical training. Plastinated organs exhibit detailed anatomy and allow for an adequate basic training prior to "in-vivo" practice . Attendees of our training course have widely accepted the use of plastinated organs due to their ease of handling and high training value. We conclude that the combination of plastinated organs and minimally invasive surgical techniques (laparoscopy, thoracoscopy , endoscopy, interventional radiology, etc.) constitutes a useful tool for both anatomical studies and basic surgical training .

The journal for the international society  for plastination. Reed RB, RW Henry. Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville , TN. USA.

After delays, the 2003 issue of the JISP has gone to press and will be presented at the 12th ISP congress. In an attempt to produce a journal of the highest quality, it was necessary to postpone the printing of JISP volume# 18 until enough appropriate articles were submitted and reviewer 's comments answered appropriately. We would like to have a minimum of five manuscripts for each volume. Manuscripts are currently being reviewed for volume # 19 of the JISP which will go to press winter of 2004 . This issue will also contain the abstracts from the 2001 interim meeting in China as well as the abstracts from the 2004 meeting in Spain. Volumes 17,18 and 19 will be submitted to Science Citation Index for consideration and inclusion in their list of scientific journals . In order to attain for our journal the respect of the scientific community , we will need to publish manuscripts which demonstrate new findings or novel ideas.

Oral presentations

A   new   method   for   preservation   of   anatomical specimens. Arnautovic  I, R Henry Avdic, V Cutahija, M Tabakovic. Department of anatomy, Faculty of Veterinary Medicine, University of Sarajevo, Sarajevo, Bosnia-Hercegovina, 1Department of Comparative Medicine, College of Veterinary Medicine, The University of Tennessee, Knoxville, TN, USA.

For hundreds of years desiccation of biological tissue has been a useful means of preservation. As well, drying of hollow organs has been an inexpensive method of preservation of anatomical specimens . However, hungry insects have a predilection for such dried organs. A mechanism to prevent infestation of the dried specimen with insects and thus prevent the destruction of the dried organs was necessary to be developed. This process has been called internal plastification and consists of two stages. 1. The preparation progresses by cleaning the organ and flowing room temperature air through the organ, as has been done historically . The organ is first dilated to the desired degree of inflation and then the exhaust port is regulated to maintain inflation. Depending on the size of the organ, drying takes three to four days. The stomach (monogastric and ruminant), small and large intestines, lungs and uterus and vagina have been preserved by this method. 2. The second phase commences after drying is completed and consists of gradual injection of plastic expanding foam, [Tekapur (Bosnia-Hercegovina) or Great Stuff (USA)].  The plastic expands in volume in all directions and hardens gradually . Depending on the product used, it enlarges two to five times. Next, more foam is injected at sites that are devoid of foam to assure complete lining and filling of the organ. The hardening time is eight hours to one or two days depending on the size of the organ . The organs filled with foam are lightweight and  the foam lining protects the internal surface against insects. The external surface is protected by varnishing and named regions may be painted for delineation of such region. This method produces a  specimen  that maintains normal anatomical structures, is durable and is inexpensive . We believe this foam could be used to dilate plastinated hollow organs  prior  to polymerization.

Ascaris plastination using the S 10 technique. Asadi MH, A Mahmodzadeh, H Haydari. Department of Anatomy, Baghiatollah University of Medical Sciences and Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Ascaris lumbricoides is one of the most common Parasites of the human digestive system. It has a world­ wide distribution  and  is particularly  very  common  in regions with poor  sanitation. Ascaris  is the largest of the common nematode parasites of the human . Different techniques are used to prepare Ascaris for educational purposes in medical colleges. The common method employing formaldehyde fixed produces specimens which are wet and with an annoying odor. The purpose of this study was to perform plastination using the Sl 0 technique . This procedure produced specimens which are dry, odorless, and easy to handle . Meanwhile, the natural shape of the specimen is preserved and specimens can be easily used for demonstration in educational courses while working with medical students.

Role of hydrogen peroxide in  plastination  of  upper and lower limbs with injection  of  colored  polymer into the arteries. Dashti GR, AR Sabahi, M Hajian, E Esfandiari, A Saki. Department of Anatomical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

Plastination is a unique technique of tissue preservation . The aim of this study was to prepare plastinated specimens of upper and lower limbs of good quality with colored polymer injected into the arteries using hydrogen peroxide to lighten muscle tissue. Materials and Methods: This study was carried out on a human body . The colored polymer was prepared by mixing colored paste with polymer , cobalt (accelerator) and peroxide (hardener). The polymer was injected into the body through femoral artery by a hydraulic pump. Dissection , dehydration , bleaching with hydrogen peroxide, defatting, impregnation and curing were carried out. Results: The plastinated specimen produced showed clearly the bright appearing muscle tissue which had fine stature,  flexibility and good  traction which was accessed by the universal test DARTC (England) apparatus in the medical physics department, school of medicine . Conclusion: The specimen obtained provided an excellent opportunity to demonstrate that hydrogen peroxide, an oxidizing agent, can play a significant role in preventing tissue from becoming dark or black in color during the course of plastination of tissue which may be due to impurities of chemicals.

Whole body plastination with  a polyester resin. Esfandiary E, M Hajian, A Rabiei. Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

In Iran due to the religious culture, body donation for dissection is very rare. There is an acute shortage of cadavers for medical schools. In  order  to  compensate for this shortage, cadavers can be preserved  with  the new preservation technique of plastination. This reality led us to find a cheap way to plastinate the specimen with a polyester resin instead of silicone. Materials and Methods: A new cheap polymer for plastination of cadavers was made and  called  P75  in  Isfahan University of Medical Sciences. The specimens were dehydrated in room temperature acetone . A vacuum chamber was made for impregnation of the specimen. Results:The product obtained was flexible, natural colored, dry and durable. This plastinated specimen is comparable with world standard plastination . Conclusions:  We  hope  plastination  techniques  become a routine work in every anatomy department of our country to save both sophisticated dissected bodies and time of the dissectors .

Refurbishment of plastinated joints for  interactive clinical teaching. Easteal R, RE Hunt, SC Pang. Department of Anatomy and Cell Biology, Queen 's University, Kingston , Canada.

In teaching of clinical anatomy it is important for students to handle human specimens. Plastinated specimens are by far the most student friendly means of affording hands-on experience to these clinical students. However, when using plastinated joints, eventually the ligaments, even plastinated ones, will tear rendering them useless for interactive teaching. Materials and methods: However , using a little ingenuity and some items found in most hardware stores, we have developed artificial ligaments for these worn-out specimens, rendering them again functional. It is important that the artificial ligaments (especially in the knee) are placed exactly in the correct anatomical location with the proper elastic coefficient. Results and conclusion : These two functions completely return the specimens to their former use as correctly operating joints, with anatomical and mechanical integrity.

India ink enhances the coloration and teaching effectiveness of plastinated specimens. Glover R, M Wells. Undergraduate Research Opportunities Program, University of Michigan Medical School, Ann Arbor, MI, USA.

The use of acetone to dehydrate tissues in the plastination process and their saturation with silicone polymer after impregnation appear to significantly interfere with permanent coloration of plastinated specimens. Therefore, it was determined to develop a method for the coloration of plastinated tissues that would withstand both of these necessary conditions. Formalin- fixed limb tissue blocks were washed, partially dried and their arteries, veins and nerves were selectively colored with several coats of India ink using a fine camel's hair paint brush. The gyri of  several brain specimens were colored in the same manner . Before of painting, as much connective tissue as possible was removed from the surface of structures to prevent India ink from bleeding color into their immediate surroundings. Painted tissues were dehydrated in acetone and checked daily for loss of color. Impregnation with silicone, under vacuum, and the application of catalyst were carried out in the usual manner. India ink colored tissues did not fade but withstood acetone dehydration, normal vacuum impregnation and curing without loss of color. Equally important, their color was not adversely affected by normal use and handling. The availability of an effective coloration methodology for plastinated specimens is desirable for many reasons. For educators, the ability to selectively color plastinated specimens should markedly improve their effectiveness m a variety of different teaching/learning settings.

Preparation and application of a new UV stabilized polyester  in  plastination   in  human  anatomy.  Hajian M, E Esfandiari, AA Rabiei. Department of Chemistry and Anatomical Department of Medical School of Isfahan University, Isfahan, Iran .

Many synthetic resins such as polyesters , silicone and epoxy resins are used for prolonged preservation of organs and tissues of man and animals without need for storage in formalin or ethanol. In this project a typical UV stabilized unsaturated polyester resin was prepared from two kinds of glycol (propylene and ethylene glycol), phthalic and maleic anhydride, a UV stabilizer ( 2-2 di hydroxyl- 4 methoxy ), a cross-linking agent (a styrene monomer) and a hardener (cobalt naphthenate). The  polyester  was  used  f0r plastination  of  a  sliced human brain and a whole body. The stages of fixation, dehydration, defatting, forced impregnation and curing were used . Important properties of the plastinated specimens, UV stability, chemical resistance, durability and mechanical properties, were investigated and compared with the specimens prepared at the institute of plastination in Germany.

Continued  studies  on  impregnation  with  silicone polymer and no additives. Henry RW, Reed  RB, Latorre K, H Smodlaka 2 Department of Comparative Medicine, College of Veterinary Medicine, The University of Tennessee, Knoxville , TN, USA, 1Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, 2Department of Anatomy and Radiology, College of Veterimary Medicine, Univeristy of Georgia, Athens, GA, USA,.

The time tested silicone technique  introduced  by  Dr. von Hagens (Biodur) uses a reaction-mixture of Sl0/S3 and has been used for impregnation and production of high-quality , aesthetic specimens for 26 years . The Biodur process is most often used at cold temperatures , but may be used at room temperature.  The  one drawback of using this reaction-mixture  is  thickening after several months. Six years ago, a reaction-mixture that is stable at room temperature was introduced. This room  temperature  process   (Dow/Corcoran)   yielded good quality specimens  and reduced  plastination  time by nearly one half. One problem of this method is a detracting granular looking film which develops on the surface of the specimens. In an attempt to decrease plastination time and have a polymer that is easily removed from fur and yet still produce the highest quality specimens, impregnation of silicone without catalyst/chain  extender or cross-linker is being  used on a variety  of specimens. Impregnation  time ranges  from one day to two weeks. The low viscosity (40 - 80 centistokes) of the polymer provides a shortened draining and manicuring time. Two regimes are used to cure the specimens: 1. Specimens are exposed to the vaporized cross-linker (S6) for 2 - 4 days then S3 (catalyst/chain extender) is wiped onto the surface of the specimen twice at 24 hour intervals. 2. Specimens are exposed to the vapor of a new Supercatalyst for 2 days. After sitting for 2 days, specimens are exposed to a volatilized chain extender (S7) for 2 days. Finally, two days later they are exposed to the volatilized cross­ linker (S6) for 4 days and allowed to remain in the closed environment as long as necessary (2 to 4 weeks) . S6 exposure is repeated if deemed necessary . Both methods result in polymerization of the outer layer of the specimen within several days. When specimens are sectioned two and four weeks later, the interior of most of the specimens contains not totally cured polymer . Some specimens take several weeks for the polymer to cure throughout the depths. Some may take months to cure. The quality of the specimens appears to be similar to that of specimens produced via the classic Biodur method . Currently, the biggest advantage is the polymer runs freely off of hair-covered specimens, which eliminates the tedious task of manicuring which is associated with hair covered specimens produced by the classic method. In time this may prove to be another useful method for producing specimens more rapidly and with out the need of refrigeration .

Plastination of  the  bronchial  tree with  RTV silicone at different developmental stages: Dehydration with alcohol at room temperature. Jimenez Mejia R, 0 Isaza Castro. Department of Morphology, Faculty of Medicine, University of Antioquia , Colombia, South America .

The difficulties associated with the process of obtaining and managing acetone in Colombia have driven a search for alternatives to the various steps of plastination , among them dehydration of anatomical specimens, which is recommended to be carried out at low temperatures with acetone. One alternative to aid in describing development of the bronchial tree from the embryonic process up to childhood stage and allow demonstration of the successive dichotomist division in the lung is by the injection of a flexible silicone. Materials and Methods : Five different ages fetuses and corpses pulmonary blocks were fixed with 10% formalin for four 4 weeks. After fixation they were dehydrated at room temperature in increasing concentrations of isopropyl alcohol (70 %, 75 %, 90 %, 95 % and  100 %) for a period  of one week in each concentration . After dehydration , the tracheobronchial tree was injected through the trachea  with  RTV silicone.  The  polymer  in  the  injected  specimens  was allowed  to  cure  for  three- days.  After  curing,  the specimens were placed in an autokey at 100°C for 30 minutes. Subsequently they were washed with water at a  high  pressure  to  remove  respiratory  tissue  and  the tissue    remains    were    manually    eliminated.    Digital images were taken . Results : This technique allowed the dehydration  of  the  specimens  and  the  injection  of  a flexible  silicone to yield  flexible  tracheobronchial  tree preparations  of  different  stages  of  development.  The developmental  progression  of the tracheobronchial  tree was readily visible in this series of lungs. Conclusions : This  work  demonstrates the efficiency  of  the dehydration with isopropyl alcohol at room temperature.   The  successful  utilization   of  a  flexible silicone  in hollow  structures  demonstrates  overcoming shortages   that   are   present   in   our   society   and   the difficulties generated partly by problems as grave as the drugs traffic .

Sheet plastination with Polyester: An alternative for all tissues. Latorre R, JM Hervas, A. Arencibia 1  Departamento de Anatomfa y Anatomfa Patol6gica Comparadas, Anatomia Veterinaria, Universidad de Murcia, 1Departamento de Metodos de Investigaci6n y Diagn6stico en Educaci6n, Universidad de Murcia, 2Departamento de Ciencias Morfol6gicas, Anatomia Humana, Universidad de Murcia, 3Departamento de Patologia Animal, Cirugia Veterinaria, Universidad de Murcia, 4I E.S.Alfonso X El Sabio, Murcia, 5J E.S. Ingeniero de la Cierva, Murcia, 6l E.S.Saavedra Fajardo, Murcia, Espana, Europe, 7 Anatomia Veterinaria, Departamento de Zootecnia, Universidade de Tras-os-Montes e Alto Douro, Vila Real, Portugal, Europe, 8Department of Comparative  Medicine, College of Veterinary Medicine, University of Tennessee, USA.

Classically, the main  application  of  polyester  polymer in plastination (P40 and P35 , Biodur™) is  the production of brain or head slices. Recently semi­ transparent body slices have been produced in our labs using P40. No literature was found on the production of body slices using P40 . The purpose of this study was to develop a protocol for using P40 to produce slices from all regions of the body. Two millimeter slices were cut from all regions of the body of a cat. The  slices were processed using  a modified  P40  technique.  Compared to the El2 method, the P40 technique offers  not  only great optical and anatomical detail, but also some extra advantages: Slices do not yellow, Ease for the beginner, and Indefinite pot life of the  impregnation  bath .  The P40 plastinated  body  slices  yielded  excellent anatomical detail of all tissues that were observed and all tissues cured in a routine manner. In addition, the slices have been excellent aids for  teaching  and research in our class rooms and laboratories.

Evaluation  of  plastinated   organs  as  a  resource. improvement   of   the   teaching-learning processes. Latorre R, MP Garcia-Sani, F Gil, M Moren0, A Agut,  JM  Quiiionero", E  Lozano5, J Herrero, F Hernandez-Pina1, H  Fenandes-  Serodio7    R W Henry8 Departamento de Anatomia y Anatomia Patologica Comparadas, Anatomia Veterinaria, Universidad de Murcia, 1Departamento de Metodos de Investigacion y Diagnostico en Educacion, Universidad de Murcia, 2Departamento de Ciencias Morfologicas, Anatomia Humana, Universidad de Murcia, 3Departamento de Patologia Animal, Cirugia Veterinaria, Universidad de Murcia, 4I E.S.Alfonso X El Sabio, Murcia, 5J E.S. Ingeniero de la Cierva, Murcia, 6l E.S.Saavedra Fajardo, Murcia, Espana, Europe, 7 Anatomia Veterinaria, Departamento de Zootecnia, Universidade de Tras-os-Montes e Alto Douro, Vila Real, Portugal, Europe, 8Department of Comparative  Medicine, College of Veterinary Medicine, University of Tennessee, USA.

Plastinated specimens can be considered didactic materials . The general goal of this investigation consisted  of  evaluating  the  use  of  plastination as a resource  towards  innovative  teaching, being  capable  of incrementing  the  quality  of  the  education-learning process in high school and university , in the region of Murcia. For this investigation, teachers and  students from three levels of education in the region of Murcia were involved : High School Education, Univeristy Education and Postgraduate Education . The decision of which plastinated specimens should be  used  was reached by consensus, in response to the needs of the different teaching levels. We worked through an interdepartmental approach. The evaluation instruments consisted of observation techniques combined with inquires and previous knowledge  and  output examination examinations. For this study, four types of instruments were selected:

  • A 3-grade  numerical  estimation  scale, registered  by the teachers, after attentive  observation  while  using the plastinated  specimens.
  • A closed  questionnaire  using  a 3-grade  Lickert-type scale, filled out by the students gathered information referring the effectiveness of the didactic resources .
  • An output examination made up in order to evaluate the quantity and  quality of the knowledge and  skills acquired as a consequence of the use of specific didactic material.

The results show that plastinated specimens are very useful as a new educative method in secondary schools and university from Murcia region. Supported by SENECA Foundation , Project:PC/2/FS/99 .

Plastination workshops:  Four  years  experience  in Spain. Latorre R, 0 Lopez-Arbors., JM Hervas, E Abel/an., JM Vazquez, M Orenes, C Sanchez, F Martinez., A Diz1 R Henry2. Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe, 1Departmennto d Anatomia Patologica Comparadas, Universidad de Cordoba, Cordoba, Espana, Europe, 2Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA.

Since 2000, the Anatomy Department of the University of Murcia has  been  promoting  plastination  techniques by conducting two-day workshops once per year. The workshops gave hands on experience primarily with the silicone cold and room temperature processes. RTV silicone and  polyester  polymers  were  also demonstrated. A maximum of 15 participants were admitted to each workshop, most of participants were from the various regions of Spain. As workshops were taught in Spanish, participants were attracted  from various  countries  of both  Central  and  South America. At the end of each workshop  all participants  filled out an evaluation sheet. Most participants  considered  these workshops a very valuable tool for them to gain the experience to begin the  plastination  technique  and  to set up a plastination lab in their University. In fact, ten (six are working and four are starting up) new plastination labs have been set up around Spain in the last five years. These additions to the plastination community are directly related to participation in Murcia's  workshops.

S-10  and   P-40   plastination   techniques   for anatomical studies of the Sea Bass (Dicentrarchus labrax, L.; Teleost, Fish).  Lopez-A/hors  0,  M"D Ayala, R Cuellar1, I Abdef , M Orenes, R Latorre. Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, 1Dept. Anatomia Veterinaria,Universidad Nacional de Aguas Calientes, Mexico, 21nstituto Oceanograjico de Mazarron, Murcia, Espana, Europe.

Plastination is very useful to study the external and internal anatomy of fish. However, its use and application is still scarce. The sea bass is an intensively reared  teleost  and  highly  appreciated  in  Mediterranean countries. Anatomical studies of this species are relevant for related histological and physiological investigations. In  this  work,  plastination  techniques were used on prosected and  serially  sectioned specimens. Two whole    specimens with lateral dissection of the body wall, two transversally sectioned and two horizontally sectioned specimens were plastinated according to the standard S-10 and P-40 techniques. The combined use of prosections and cross­ sections was very useful to describe the gross anatomy of sea bass. Cross-sections at the level of the head allowed the study of internal organs which are difficult to view by dissection: encephalon, eye-ball and related structures. Transverse and horizontal sections allowed a detailed description of the topography of the viscera. The P-40 thin sections allowed evaluation  of anatomical structures at submacroscopic level. A common problem with the S-10 technique was that the muscle tissue tended to display a final orange appearance, which may be due to the high fat content of farmed fish.

Curing influences on tissue preservation of silicone plastinated organs. Lopez-A/hors O, F Gil, M Orenes, M"D Ayala, R Henry , R Latorre. Departamento de Anatomia y Anatomia Patologica Comparadas, Anatomia Veterinaria, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe,  Department  of  Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA.

Deplastination of silicone plastinated specimens  has been proposed as a valid technique for histological studies. Despite some structural changes deplastinated/paraffin  embedded  tissue  maintains  most of its structural features. Fixation and  deplastination have been designed as the most critical steps when plastination is used for histological purposes. However, the influence of the curing on the tissue structure of plastinated organs has not been evaluated yet. With this objective, heart, lung, kidney,  liver,  uterus,  omasum and rumen of several  domestic  mammals  were plastinated according to three standardized protocols: Biodur S-10, Corcoran PR-10 and  Vis-Docta  HS I. Tissue samples of these organs, before and after curing were deplastinated with 5% sodium methoxide in methanol for 48 h.  Afterwards  the  samples  were washed in methanol, routinely processed for paraffin embedding and sectioning and finally stained with hematoxylin-eosin. Results showed that the  curing process  influences  the  tissue  preservation,  however with no significant differences between the three plastination techniques. Uncured samples had little or unappreciable tissue damage, whereas cured samples showed histological change. To find out whether  these findings are only produced by the curing process or also related to other processing steps such as the deplastination methodology or the paraffin embedding deserves further studies.

Fine configuration of the connective tissue in the posterior atlanto-occipital region: An E-12 sheet plastination and confocal microscopy study. Nash L. Clinical Anatomy Lab, Department of Anatomy & Structural Biology, School of Medical Science, University of Otago, Dunedin, Otago, New Zealand.

Aberrant kinematics of the dura-muscular and dura­ ligamentous connections located in the posterior atlanto-occipital (PAO) region are widely thought to transmit mechanical forces  from  the  intervertebral joints to the pain-sensitive spinal dura generating pathologies such as cervicogenic headaches . However, little information exists regarding the anatomical arrangements of the main structures, i.e. rectus capitis posterior minor (RCPm), PAO membrane , nuchal ligament, and spinal dura. The aim of this study, using plastination and confocal microscopy , was to examine the morphology of the fibrous structures and  to determine the anatomy of the dura-muscular and dura­ ligamentous connections in the  PAO region . Materials and Methods: Twenty-two embalmed adult human cadavers were used. Thirteen were prepared by sheet plastination. Plastinated tissues were examined macroscopically using of a light box and dissecting microscope  (xl .25)  and  microscopically  with  a confocal microscope. Two dissection procedures were performed on nine cadavers. The first involved conventional mid-sagittal sections from the  occiput  to C7. The second entailed a lateral approach in the atlanto-occipital region . Results : Direct evidence from cadaveric and plastinated material revealed: 1. Anteroinferiorly orientated tendon fibers from  the medial and deep aspects of RCPm  directly  connected with the spinal dura within the PAO  region.  2 .  The POA membrane was formed by the RCPm fascia and vertebral vascular sheath and inferiorly was fused with the spinal dura. 3. No direct connection between the nuchal ligament and the posterior cervical dura matter was found in the PAO region. Conclusions: The deep layer of the RCPm fascia and the perivascular sheath of the external vertebral plexuses are the main connective tissue structures of the PAO membrane between RCPm and the spinal dura in the  PAO  region.  The morphological features of the RCPm tendon and fascia indicate that they have an important role in the maintenance of posterior craniocervical stability, the prevention of the dural infolding during extension, and the generation of cervicogenic headaches.

Plastinated organs  as  models  for  endoscopic training. Oppitz M, A Kirschniak, U Drews . Department of Experimental Embryology, Institute of Anatomy, University of TU.bingen, Tubingen, Germany, Europe.

For training medical doctors skills for endoscopic surgery, a variety of procedures and equipment have been established. Proposals range from artificial phantoms to surgery training on animals. However , these procedures have the disadvantage of lacking the normal human topographical anatomy. In our clinical anatomy training unit, cadavers for training surgical endoscopy were preserved . Apart from whole body preservation with ethanol/glycerol and fixation after Thiel, plastinated organs of the gastrointestinal (GI) tract are used for training of flexible endoscopy. Materials and Methods : Phantoms were prepared from formalin-fixed human cadavers using classical anatomical and appropriate surgical techniques. Blood vessels were filled with colored silicone. After fixation of organs and dehydration in acetone at -20°C, the preparations were impregnated with silicone for 4 to 8 weeks depending on size and left to polymerize for 4 to 7 days. Results: The procedure described produced plastinated organ preparations of  the  gastrointestinal tract that  simulate  clinical  human  endoscopic  anatomy in a close-to-reality setting.  The  disadvantage  of reduced tissue softness in plastinated organs is compensated by the realistic detail such as excellent visualization of the course and topography  of the blood supply as seen from  the  endoscopist 's  view. Conclusion: Endoscopic training  with  realistic simulation of anatomical detail is important for medical personnel when having  to  deal with  emergencies  such as upper GI-bleeding of peptic ulcers or esophagus varices. In addition to hands-on experience, the detailed knowledge of anatomical detail gained from endoscopy of plastinated organs will be of great help for stopping GI- hemorrhage.

Enhancing the  educational value of plastinated specimens in the gross anatomy curriculum: Adding color to human neurovascular pathways. Raoof A, C Parres, J Blythe. Anatomical Division, Medical Education, University of Michigan Medical School, Ann Arbor, MI, USA.

In recent years, plastinated specimens have become invaluable assets to the gross human anatomy curriculum , furthering the development of students' identification and diagnostic skills of anatomical structures. Students often have difficulties, however , in conceptualizing important neurovascular pathways and relationships using these virtually colorless specimens. An applicable method for improving plastinated specimens as learning tools involves the use of acrylic paints. Nerves, arteries, and veins of plastinated specimens are actually painted by hand, yielding detailed, colorful specimens and providing a more holistic representation of the neurovascular picture. Methods: Plastinated upper limb, brachial  plexus, pelvis, and two lower limb specimens were selected for coloring. Nerves, arteries, and veins of each specimen were cleaned using dissection tools. Each nerve/vessel was carefully painted the appropriate color and then allowed to air-dry  for  twenty-four  hours. Questionnaires regarding the effectiveness and applicability of the colored specimens  were administered to eleven students enrolled in a 2003 summer pre-matriculation anatomy course at the University of Michigan . Results: Life-like , durable, and anatomically accurate models representing in  color major neurovascular pathways in human upper and lower limbs were created for use in the gross anatomy medical education curriculum . Questionnaire responses were overwhelmingly positive - 81.8 % of the students supported that "the painted specimens would reinforce the value of dissection and would be beneficial in a teaching environment." Discussion : Colored plastinated specimens proved to be effective and engaging learning tools for medical students. If put to use in the gross anatomy curriculum , these specimens would lend to a fuller understanding of neurovascular relationships and of important physiological processes of the body. A few problems to work through: Time-consuming work to hand-paint  the  specimens  and  more  experimentation should be done to provide  better paint  durability over time. Advantages of this coloring technique over others such as casting: Specimens chosen for casting are usually hollow visceral organs while our coloring technique is applicable to almost any specimen.

Ultrasonographic anatomy of the horse temporomandibular joint: A study by P-40 sections. Rodriguez   MJ,  A  Agut,  F   Gi1,  M  Rivero2     E Martinez1, R Latorre 1 Medicina y Cirugia Animal , Facultad de Veterinaria, Universidad de Murcia, Espana. 1Anatomia y Embriologia , Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana . Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Las Palmas, de Gran Canaria, Espana , Europe.

The aim of our study was to establish a protocol of the ultrasonographic evaluation for the equine temporomandibular joint  (TMJ), to describe the normal macroscopic and ultrasonographic anatomy with cross­ sections made on the same planes. Materials and Methods: Twenty-four TMJ from twelve horse heads were used for ultrasonographic studies. The dorsal synovial pouch was injected via its caudolateral compartment. Specimens were frozen at -80°C and sectioned with a high-speed band saw at desired thicknesses (2 mm). The cut surfaces of each section were rinsed and numbered before fixation. Photographs of both surfaces of each section were taken before and after fixation. Sections were plastinated using the P-40 technique . Results: Our ultrasonographic and anatomic results suggest that three sonograms from lateral, rostral and caudal approaches are necessary to provide  a correct and complete ultrasonographic evaluation of the equine TMJ. The structures that could be  evaluated were the articular surfaces, articular cartilage in yearlings, articular disc  and  its  caudal fibrocartilaginous expansion, intra-articular fat tissue or retro disc tissue,  caudolateral  compartment  of  the dorsal synovial pouch, articular capsule, parotid gland and subcutaneous tissue . However, the ligamentum laterale and ligamentum caudale, the vascular  and nervous structures and the ventral synovial pouch could not be identified . Conclusion: The use of the anatomic cross-sections made on the same planes as the sonographic views in the equine TMJ allowed correct identification of all structures and established a direct correlation with other adjacent structures.

Clearing and plastination of anatomical sections: An alternative method to study the murine vascular system. Ruberte  J, C Llombart,  M  Navarro,  G Ramirez 1 MD Ayala 1  R Latorre1. Unidad de Anatomia y Embriologia . Facultad de Veterinaria. Universidad Aut6noma de Barcelona, Espana, 1Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe.

Murine blood vessels are too small to study by dissection, but too large for satisfactory histological work. Injection of blood vessels and clearing the tissues have  been  classical  methods  to  analyze  small  vessels (Tompsett, 1970). However, long term  exposure to vapours can be hazardous, when the  Spalterholz clearing technique (1914) is used . Moreover , preservation of  specimens  requires  much  attention, when glycerine is used for transparency.  The  aim  of this work was to preserve cleared sections of mice with sheet plastinatiun techniques, in order to avoid toxicity and conservation problems. Materials and  Methods : after intravascular injection of colored latex through the

thoracic aorta, mice were frozen at -80°C and sectioned transversely with a high-speed band saw at desired thicknesses (2-4 mm). Two different techniques were used for clearing. With the Spalterholz technique, after dehydration, impregnation with methyl salicylate and benzyl benzoate was necessary for a  complete transparency of the tissue. With the glycerine technique (Staples and Schnell, 1964), initial clearing was accomplished by soaking the sections in an aqueous solution of potassium hydroxide; after selective staining of calcified bones with Alizarin Red, complete transparency was obtained  by immersion  in a mixture of glycerine and benzyl alcohol. Sections were plastinated using the P-40 and E-12 techniques. Different dehydration protocols were used before impregnation. Results and Conclusion: The best results were obtained in Spalterholz cleared  sections impregnated in E-12, following the regular protocol of the technique , without previous steep in acetone. P40 impregnated sections  showed  less  transparency  than E12 impregnated sections. A proper adaptation of the sheet plastination technique  allows  preserving  of cleared sections.

Plastination and animation: Pedagogic-didactic proposal for study of fetal circulation in ruminant fetuses,  as  a   paradigm   for  the   study  in  other mammals. Serodio HC, J  GouveiaGif , G Ramirel, R Latorre3. Anatomia Veterinaria, Departamento de Zootecnia, Universidade de Tras-os­ Montes e Alto Douro, Vila Real Portugal, 1Metodologia de Interven<;il.o Educativa, Esco/a Superior de Educa9iio Paula Frassinetti , Porto . 2Departamento de Engenharia BiolOgica e Ambiental, Universidade de Tras-os-Montes e Alto Douro, Vila Real Portugal, 3Anatomia y Embriologia , Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe .

Reasons that justify teaching of fetal blood circulation to veterinary and medical students are: 1. Analysis of patterns of venous blood flow in fetal vessels, as a way of detecting fetal distress;  2.  Interpretation  of congenital malformations associated  with the retention of fetal circulatory structures; 3. Study of the effect of drugs which combat these malformations. Based on the need of pedagogic methods  that  allow  understanding (as opposed to simple memorization), self-studying, as well as heightening the interest of the subject, the authors conceived a pedagogic-didactic sequence designed to be used in the classroom . Students, who intend to work and develop their own knowledge , have free-access to this resource. Material and Methods: Two animations  were  developed  using  Macromedia Flash™, two self-study guides and plastinated bovine fetuses (Biodur® S10). The  pedagogic  sequence consisted of  previous observation  of  an  animation, which shows the trajectory of circulating blood on a photograph, followed by a  second  animation  that invites the student to fill in an equivalent diagram, and presents new information as it compares  photographs with echographic images. Additionally, the student has access to fetuses, specifically dissected and plastinated for this purpose. Results and Conclusions: The first animation gives the student  a reasonable perspective of the topographical anatomy of  the  dissected  fetus  and the variation of blood oxygen and nutrients at different sites in the body . The second animation allows the student to consolidate the concepts presented in the first animation and expand them, as it compares the anatomy of newly presented anatomic structures with those of echographic images . Plastinated fetuses demonstrate significant morphology and are considered crucial resources. They allow manipulation and three­ dimensional  topographical  analysis.  Fetuses  bring reality to the subject, demystifying  it  and  developing the student's self-confidence as they experience anatomical facts that will serve as basic knowledge for future problem solving.

Comparison   of   plastinated     specimens   prepared using five silicone polymers and five techniques. Smodlaka H, RW Henry1, RB Reetf , R Latorre2 0 Lopez-Albors2, JM Hervtis2, R Cuellal. Department of Anatomy and Radiology, College of Veterinary Medicine, Univeristy of Georgia, Athens , GA, USA,1Department of Comparative Medicine, College of Veterinary Medicine, The University of Tennessee, Knoxville , TN, USA, 2Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe. 1Departmento Anatomia Veterinaria. Universidad Nacional de Aguas Calientes, Mexico.

The impregnation technique for  plastination  of biological tissues has remained  nearly  the  same  since its inception twenty-six years ago. The polymers, cross­ linkers, chain extenders and catalysts are all products currently used in the silicone polymer industry. The major differences in the  Biodur,  China,  Corcoran, Italian and North Carolina A (NCa) and B (NCb) processes is in how the components are combined: Biodur, Italian and NCa combine polymer, catalyst and chain extender in the impregnation bath and after impregnation cross-linker is applied. China and  NCb uses  only the polymer  and later  a catalyst/cross-linker may be added. Corcoran combines polymer and cross­ linker and later the catalyst is added. Specimens were prepared using all six recipes and techniques in order to compare the final product.  Specimens will be presented at  the  12 hour International Congress  on  Plastination  for evaluation. Results will be presented  at the conclusion of the congress.

Sectional anatomy of dog  knee  joint:  A  study  by MRI,  Ultrasonographic   and   plastinated   sections. Soler M, A Agut, F Gi/1, JM  Viizquez1, A Arencibia 2 R Latorre 1 Medicina y Cirugia Animal, Facultad de Veterinaria, Universidad de Murcia, Espana. 1Anatomia y Embriologia , Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana. 2Anatomia y Embriologia , Facultad de Veterinaria, Universidad de Las Pa/mas de Gran Canaria, Espana, Europe.

The purpose of this study was to use a combination of P-40 plastinated slices, ultrasonographic and magnetic resonance (MR) images to better understanding the anatomy of the normal canine stifle joint. Materials and Methods:  Ultrasonographic  study was performed  using a 7.5 MHz linear  transducer.  Magnetic  resonance images of the stifle joints were made in sagittal, dorsal and transverse planes using a 0.5 Tesla GE magnet and a solenoidal human extremity coil. A Tl-weighted spin echo and T2*-weighted gradient echo images sequences were used . The canine stifle specimens, were frozen at - 80°C after intraarticular injection with latex and sectioned either transversely or sagittally with a high­ speed band saw at desired thicknesses (2-3  mm).  At least twenty sections  were  obtained  from  each specimen. Both surfaces of each  section  were  rinsed and photographed  before  and  after  plastination. Sections were plastinated using the P-40 technique. These plastinated slices were used to facilitate an accurate interpretation of the anatomical structures. Results: The normal anatomic structures that could be consistently visualized ultrasonographically included: normal cartilage on the femoral condyles  and  the trochlear ridges; the patellar ligament that  was hypoechoic with  a hyperechoic peritendon ; medial and lateral menisci , cranial and caudal cruciate  ligaments, and the tendon of the long digital extensor muscle all of which were hypoechoic compared to the patellar ligament. MR images showed ligaments, tendons and menisci that were seen with a low intensity signal. Articular cartilage had a similar low signal in TI- weighted SE sequence, but a very high intensity signal in T2*-weighted gradient echo images. The various plastinated  anatomic sections were used to identify the structures of the stifle joint as well  as  correlate anatomic  structures  with  the   MR   images. Conclusions :The combination of sheet plastination and imaging  techniques  as  described  in  this  study  allow students,  radiologists   and   anatomists   to   gain   better insight into the three-dimensional relations of anatomic structures of certain regions, in this case  the  canine stifle joint. Sheet plastinated anatomic sections are a valuable tool to teach the interpretation of the ultrasonographic and MR images of the canine stifle joint.

High  temperature  in  plastination.  Sora  M-C,  B Strobl. Plastination Laboratory, Center for Anatomy and Cell Biology, Medical University of Vienna, Vienna, Austria, Europe.

The E12 method of plastination is usually used to create 2.5 - 5 mm transparent slices. If thinner slices, 0.5 - 1.5 mm, are needed, it is necessary to  use  the epoxy thin-slice plastination method. By using this method, the specimen is first plastinated as a block and then cut in thin slices. The impregnation  temperature is the key element in order to obtain  complete impregnation of the desired block and contrary to all other plastination methods uses high temperatures. The main goal of our presentation is to describe the use of high temperature for processing  1  mm  epoxy plastinated slices. Only by using high temperature is the polymer thin enough to penetrate into the middle of the processed specimen.

P35 Plastination Experiences with delayed impregnation.  Uzel  M,  A  Weiglein1Vocational School of Health Services, Istanbul University, Istanbul, Turkey, 11nstitute of Anatomy, Medical University Graz, Graz, Austria, Europe .

During an educational demonstration of P35 technique , brain slices which had stayed immersed in P35 resin in a cooling room (5° Celsius) for approximately  two years were used . During the procedure, it was observed that the resin was quite sticky and it was very difficult to take the steel basket with the brain  slices from  it. After removal of the basket from the immersion bath, the:  a) Slices were rigid, fragile and difficult to handle, b) Filter paper between the slices was almost united with the slices, and c) Some resin remnants stuck onto the metal Because of  their fragility,  it  was difficult to put the slices onto the glass plates to set up the double glass chambers without the slices breaking. After the double glass chambers were assembled , the chambers were filled with P35/A9 mixture and slice position   was  corrected   with   a  wire.   The  hardening procedure was the usual UVA-light - heat combination for P35 . Despite the long immersion period and the problems encountered with removal of the slices, the final result was satisfactory from an optical point of view. As a conclusion, the time for the production of brain slices may be extended up to several months by leaving the slices in the cold immersion bath; however , periods of too much length need to be avoided because the slices start to cure where they are in direct contact with the steel grid.

Plastination and sectioning of the cranio-cervical junction. Weninger B. Institute of Anatomy, Medical University Graz, Graz, Austria, Europe.

The aim of this study was to visualize aberrant joints between the occipital bone and the dens of the axis and to preserve them in a stable form.  Materials  and Methods: Heads with necks still attached were used and preserved after Thiel. To  determine  which  heads showed the  desired  characteristics ,  CT-scans  were made of the atlanto-occipital region in advance and heads were selected  accordingly.  The  heads  were placed in a -70° C freezer. Of two  frozen  heads, one with latex injected vessels  and  the  other  without, squares with edges about 3 cm long were cut, which contained the occipital  condyles, the atlas and the axis. These squares were dehydrated in cold acetone (-25°C). Subsequently , the squares were plastinated using E12. The cured squares were then cut with a diamond-wire saw. Results : The plastinated squares could be cut into very thin slices without much loss of material and with the advantage of reaching the desired plane of section gradually and exactly under superv1s1on. After appropriate sectioning, the joint was clearly visible and stabilized for further examinations . Conclusions : The technique described is a valuable  means  of visualizing the structure of joints in sections without  the disadvantage of dislocating the head and  cavity of the joint , be it normal or - as in this case - an aberration .

Posters presentations

Nervous  tissue  -  Plastination  versus  Embalming: The UKM experience. Azian AL, AG Norzana, HS Farihah, AR Fairuz, G Hairi, 0 Faizah. Department of Anatomy, Medical Faculty, Universiti Kebangsaan Malaysia.Kuala Lumpur, Malaysia.

Anatomical study requires long-lasting preservation of the cadaver . Arterial embalming which has been used since the 17th century allows for a three-dimensional study  of  the  human  body .  However,  in  a  tropical climate, the cadavers need more meticulous care  and they require space for storage tanks. As well, formalin may pose as an occupational  hazard.  Due  to  the shortage of cadavers, specimens  need  to  be  preserved for a longer duration while retaining its natural and realistic  appearance . With the advent of plastination  as a tool for research and teaching, some of the problems encountered with embalming can be  overcome. However, plastinated tissues at the UKM have some shortcomings when compared to embalmed specimens. Comparisons were made between brain and spinal cord specimens either whole or cross-sectioned  preserved using the embalming solution only and plastination post-embalming. Results  showed  that  brain  tissues using the plastination technique shrunk between 30  - 50% compared to the arterial embalmed specimens. These plastinated tissues were hard, brittle and not pliable enough to show the sulci and gyri of the brain. Blood vessels and cranial nerves looked wiry and broke easily. Cross-sectioned specimens were darker in color, thus rendering it difficult to differentiate between gray and white matter. Apart from shrinkage, spinal cord specimens fared better and structures were maintained using plastination techniques.  Various  factors contributed to the poor  results  including  specimens being prepared at room  temperature , improper concentration of acetone during dehydration (final concentration of < 100%), insufficient  dehydration period, a higher concentration of hardener ( 1%) and inappropriate pressure during the forced impregnation process. Based on these experiences, steps have been taken to rectify the mistakes and improvement of the techniques involved in order to  obtain more favorable results .

Plastination of head and neck,  posterior  wall  of trunk, and spinal cord with injection of colored polymer into the arteries. Dashti G, A Sabahi, H Ghaffary. Anatomy Department, School of Medicine, ls/aha University of Medical Sciences, Isfahan , Iran .

The aim of this study was to prepare plastinated specimens of the head and neck, posterior wall of trunk and spinal cord by injecting colored polymer into  the arteries . Materials and methods : After fixation of a human cadaver, colored polymer was injected into the femoral artery by the hydraulic pump . After a few days dissection, dehydration, defatting, impregnation  and curing were carried out. Results : Plastinated specimens obtained were comparable with specimens prepared in Heidelberg, Germany , for their stature, flexibility and traction as done by the universal test Dartc(England) apparatus.   Conclusions:   According   to  this   study,  p-value was greater than 0.05, which shows no significant difference statistically. The specimens obtained were clear, dry, safe, durable and provided an excellent opportunity to demonstrate and study  the  dissected areas.

Plastination of fungi as an aid in teaching botanic classification. Diz A, A Martinez -Galisteo, M Sanchez-Rodriguez1, A  Conde-Perez2. Department of Comparative Anatomy and Pathology. Faculty of Veterinary Sciences. University of Cordoba, Espana, 1Department of Animal Production, Faculty of Veterinary Sciences, University of Cordoba, Espana, 2Sanitary District Jaen Sur. Andalussian Health Service, Junta de Andalucia , Espana, Europe.

Picking fungi is a growing hobby in south Spain. When eating is the intended purpose, greater security is necessary to avoid intoxication including  death.  The aim of this study was to preserve fungi picked during the different seasons of the year, and make a collection of the more frequently collected fungi of south Spain which would aid in teaching amateur people the botanic classification of fungi in the growing local Mycological Societies. Materials and methods: Country fungi were picked in the various stages of growth. The fungi were frozen as rapidly as possible to  maintain  their  shape until transport to the lab. A  GPS location detector was employed to localize the point where each fungus was picked. The standard S-10 silicone  technique  was applied for permanent preservation of  fungi.  To preserve color, no species were mixed during dehydration. The fast gas curing method was used to try and minimize shrinkage. The fungi were classified and placed in padded boxes to facilitate transport . Results: Fungi were successfully plastinated using the S 10 method. Our results were mixed and some  problems were identified : Different degrees of shrinkage, shape changes, and discoloration . Plastination is  a  valid method to preserve fungi collections for teaching mycological classification all year long. However , the current results suggest that they are best used as complementary material to fresh, photographic and bibliographic sources. Conclusion: Plastination of fungi with the standard S-10 silicone technique is a valid method of preservation of fungi and has become a well accepted method of preservation of fungi by local Mycological Societies. Plastinated fungi are used to complement the teaching  of the classification  of fungi to amateur people. However, improved techniques must be found for our plastination lab in order to obtain the better results.

Plastination  of  exotic  animals  in   veterinary medicine. Diz A, J Vivo, F Miro, JL Morales, JM Molero. Department of Comparative Anatomy and Pathology. Faculty of Veterinary Sciences. University a/ Cordoba, Espana, Europe.

Plastination is used more and  more  every  day  around the world as a process for preservation for whole body, body parts and organs of domestic animals in veterinary medicine . In developed countries, it  is  more  frequent use exotics animals as pets. Anatomical and clinical knowledge is growing  daily in veterinary schools. The aim of this study was to preserve  by  silicone plastination reptiles ,  amphibians,   fishes  and invertebrate animals. These zoological  groups  are known as exotic species in veterinary medicine . Materials and methods: Specimens were acquired from various places (necropsy rooms, markets, and dead animals found in the soil). This material was plastinated using the Biodur S-10  standard  technique  develop  by Dr. von Hagens. Results:  Reptiles,  amphibians  and fishes are specimens which are preserved well by this technique . Although those with vivid colors may result in loss of coloration to varying degrees. Moreover , they may shrink depending  on  their  structure  and consistency. Shrinkage can be reduced by injecting polymer-mix from the forced impregnation bath  by mouth, anus and corporal cavities. Plastination of invertebrates had similar results although it is important to consider the great diversity of this zoological group. Conclusion: Plastination is an appropriate method to preserve zoological specimens different  from mammalians . This technique allows the formation of collections of exotic animals, from both healthy and pathological  origins, for study in veterinary  medicine.

Some aspects  of  fungi  plastination.  Diz  A,  A Martinez -Galisteo, J Berlango 1 A Conde-Perei. Department of Comparative Anatomy and Pathology. Faculty of Veterinary Sciences. University of Cordoba, Espana, 1Reina Sofia University Hospital. University of Cordoba, Espana, 2Sanitary District Jaen Sur. Andalussian Health Service, Junta de Andalucia , Espana Europe.

The aim of this study was to determinate if plastination is a valid preservation technique to obtain permanently preserved fungi as an aid to its botanic identification . Materials and methods: Fresh fungi were picked in the Sierra Morena hills, selecting young and healthy specimens. They were frozen and maintained in a household deep freezer until transport to the lab. Fungi were plastinated following the standard  S-10 silicone technique without fixation . Before dehydration, fungi were arranged loosely into an acetone 1resistant plastic basket and covered with a polymer mesh which was fixed to the basket to prevent floating. Dehydration was made in successive (3-4) baths of 100% cool acetone (- 250C) until no more than I% residual water was present. Great care must be used in not mixing different species to preserve color. After dehydration, forced impregnation of S 1O/S 3 was carried out. After impregnation and prior to curing, the fungi were placed in a vertical position (12-24 h.) in the freezer to drain the excess polymer. The fast gas curing method was applied to reduce shrinkage. Specimens were wiped of oozing polymer throughout the curing process. Results: Plastinated fungi, with great interspecies differences, discolor and shrink to some degree depending on the variety of the fungus. Nevertheless , they are preserved permanently , permitting identification of poisonous and edible fungi at any moment. Conclusion : Plastination of fungi with the standard S-10 silicone technique  is  a very valuable method for fungi preservation , although shrinkage, shape changes and partial damage of colour is observed .

Computerized reconstruction of  a  plastinated human kidney using serial tissue sections. Doll F, S Doll,   M-C   Sora1 Lozanoff. Department of Biomedical Sciences, University of Hawaii, School of Medicine, Honolulu, HI, USA, 1University of Vienna, Vienna, Austria, Europe, 2 University of Saskatchewan , Saskatoon, SK, Canada.

Baskground: Computerized  reconstruction  of anatomical structures  is becoming very useful for developing anatomical teaching modules and animations. Although databases exist comprising serial sections derived from frozen cadaveric material , plastination represents an alternate method for developing anatomical data useful for computerized reconstruction . The purpose of this study is to describe a method for developing a computerized model of the human kidney and ureter using plastinates. Materials and Methods: A human kidney was obtained, plastinated , sectioned and subjected to 3D cmputerized reconstruction using WinSURF modeling system (SURFdriver Software). The kidney was generated rapidly and rendered easily on Windows laptop machine in real time. Results: Qualitative observations revealed that the morphological features of the model were consistent with those displayed by typical cadaveric specimens. Morphometric analysis indicated that the model did not differ significantly from a sample of cadaveric specimens. Conclusion : Although further experiments will be required to mm1m1ze tubular compression of the ureter, these data support the use of plastinates for generating tissue sections useful for 3D computerized modeling . Supported by BRIN NIH Grant Number RR-164676 .

Latex injection for the arterial vascular patterns of the Ethmoidal arteries. Erdogmus S, F Govsa. Ege University Faculty of Medicine, Department of Anatomy, Ege University, Izmir, Turkey.

Ethmoidal arteries are damaged in endonasal surgical interventions and in operations performed on the inner wall of the orbit. Ligation of the anterior and posterior ethmoidal arteries causes severe nasal bleeding and mortality . The aim of this study is providing ligation with the appropriate intervention thus reducing the rate of damage to these arteries. Materials and methods: Vessels supplying blood to the orbit were examined by dissecting 16 cadavers that were fixed with a 10% formaldehyde solution. Red latex was injected into the internal carotid arteries before dissection. The size and distribution of the vessels were investigated and the primary supplying vessels were identified. Results : The anterior ethmoidal artery was observed in all specimens except one. The diameter of the artery, 0.92 ± 0.2 mm on the right and 0.88 ± 0. 15 mm on the left, was thicker than the posterior ethmoidal artery. The branching of the anterior ethmoidal artery from the ophthalmic artery was determined to be four different types. The posterior ethmoidal artery was observed in all  cases.  The diameter of the posterior  ethmoidal  artery was 0.66 ±0.21 mm on the right and 0.63 ± 0.19 mm on the left. Conclusion: Knowledge concerning the  variations  in the anatomy of the ethmoidal arteries is important for surgeons. This data emphasizes the importance of a sound anatomical understanding of the arterial system of the ethmoidal cells  in relation to the surgical procedures to be done.

Plastination of musculoskeletal system: A useful tool in teaching. Faizah 0,, HS Farihah, AR Fairuz, G Hairi, AL Azian. Department of Anatomy, Medical Faculty, Universiti Kebangsaan Malaysia, Kuala Lumpur , Malaysia.

Interest in the human body leading to the dissection of corpses has been documented sine the 15th century. Although the study of cell has reached molecular levels, macroscopic anatomy remains as  an indispensable component of the medical training .  The musculoskeletal system being the backbone of Orthopedic Surgery need to be learnt thoroughly in the early  years  of the  course.  Pictures  and  images  alone may be inadequate to achieve this objective. Hands on experience is still seen as one of the best methodology in learning the musculoskeletal system. However, wet and smelly specimens deter effective learning. Plastination techniques allow specimens to be kept longer, dry and easy handling with minimal problems of storage and transportation. Regions of the upper and lower limbs were carefully dissected  to reveal superficial and deep structures. These dissected parts were then processed according to standard plastination procedure which is dehydration, forced impregnation and gas curing. Muscles tendons, cartilages and bones plastinated with silicone rubber retained their natural appearance and colour within the acceptable limits of shrinkage. Although the consistency were rather hard, the results obtained were comparable to the exhibits shown in Body Worlds. However, comparing to the formalin    preserved    specimens,   the    neurovascular bundle were more brittle . Plastination technique provided  more  opportunity  for  students'  self-directed learning since specimens are easily accessible, dry and odourless. In order for the specimens to achieve more flexibility, the concentration of the hardener has been reduced. Steps in dehydration and impregnation are strictly followed in order to obtain specimens which looked natural and realistic and at the same time more durable in view of shortage of cadaver in the near future.

Pitfalls of plastination: Solid and hollow organs. Farihah S, 0 Faizah, AR Fairuz, G Hairi, AL Azian. Department of Anatomy, Medical Faculty, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.

In teaching Gross Anatomy , internal organs are an invaluable teaching aid especially when cadavers are becoming increasingly difficult to obtain. Current methods of preservation , embalming with a solution with formaldehyde as the main preservative, exposes individuals to a variety of hazardous materials and causes irritation to mucous membranes of the eyes and respiratory tract. Embalmed organs remain wet with a fairly strong odor of formalin and there is a tendency for fungal growth in a humid environment. The plastination technique of silicone impregnation allows preservation of organs making them durable, dry and odorless thus facilitating handling. However, a number of technical problems were encountered with the plastination method used in the UKM Medical Faculty. Solid and hollow organs which were plastinated include the liver, lungs, heart and stomach. Results showed that all the organs shrunk between 10 - 30% with hollow organs shrinking more when compared to solid organs. Lungs and stomachs appeared unnatural as they collapsed and become hardened. Plastinated organs from embalmed cadavers were darker in color. Chambers of the heart often contained hardened polymer and were collapsed and difficult to visualize. Multiple technical errors have been identified which gave rise to these results : 1. Improper temperature (warm) and concentration of the acetone during dehydration , 2. Use of a higher concentration ratio of chain extender to polymer, 3.  Inappropriate  pressure and duration during forced impregnation , 4.Hollow organs were not inflated at the beginning of the plastination process. Hydrogen peroxide may be used to decolorize darkened specimens following the initial embalming process. Other corrective measures have been taken to improve the quality of the specimens.

Basis   of   latex   injection   techniques   in   arterial features of the posterior maxillary segment  in relation  to the Le Fort I osteotomy.  Govsa F 1 2 Zuhaf , Y Pinar1. 1Ege University Faculty of Medicine, Department of Anatomy, and 2Ataturk Health Care College, Ege University, Izmir, Turkey.

The most common site of hemorrhage in maxillary osteotomies is the posterior maxilla. Better understanding of the vascular anatomy in posterior maxilla may minimize vascular complications . The aim of this study was to investigate the posterior maxillary region and establish safety guidelines for the Le Fort I osteotomy . Materials and methods : Intra-arterial injection of colored latex and dissection of the vessels running to the palate was performed in 30 male cadavers. The size and distribution of the vessels were investigated and the primary supplying vessels were identified. Results : The maxillary artery enters the pterygopalatine fossa approximately 16.2 mm superior to the nasal floor and gives off the descending palatine artery. The descending  palatine artery travels a short distance within the pterygopalatine fossa and then enters the greater palatine canal. It  travels approximately 10 mm within the canal in an inferior, anterior and slightly medial direction to exit the greater palatine foramen in the region of the second and third molars. Conclusion : Injury to the descending palatine artery during Le Fort I osteotomy can be minimized by not extending the osteotomy more than 35 mm posterior to the piriform rim in males. Pterygomaxillary separation should be made by closely adapting the cutting edge of a curved osteotome or right-angled saw to the pterygomaxillary fissure while  avoiding excessive angulation. Furthermore, the superior cutting edge of the osteotome or saw blade should be less than 10 mm superior to the nasal floq. This study establishes that while it is possible to ' conserve the descending palatine arteries intact during Le Fort I maxillary osteotomies, injury to them leads only to moderate and transient ischemia of the palate thanks to vascular substitution which comes from the ascending palatine arteries and pharyngeal branches running through the soft palate.

Starting plastination  in the 2000's.  Guintard C, E Betti,  J-C  DesFontis,   G  Grondin1 Vi!terinaire de Nantes, Unite d'anatomie comparee, Nantes, France Europe, 1Universite du Quebec a Trois­ Rivieres,   Departement    de   chimie-biologie,    Trois­ Rivieres, Qc, Canada.

Since the introduction of the technique in 1979 by Dr. von Hagens, plastination laboratories have been developed around the world. Each time a project for a new laboratory was presented, two important questions were raised: What equipment is necessary and how much will it cost? These two questions still appear today when someone decides to start plastination. The design and starting cost of plastination laboratories have been reported in the literature in past years (Lischka and Prihoda, 1987; Gubbins, 1990; Bore and Boyes, 1994; O'Sullivan and Mitchell, 1995; Briggs et al., 1997) but has not been updated since 2002. The present work reports on the starting of a plastination laboratory in the summer 2003 at the "Ecole Nationale Veterinaire de Nantes" with a listing of the equipment purchased for this project and its cost. The selected location for this laboratory will be described along with the modifications that were necessary to make the laboratory safe and functional. The cost of these modifications is also listed.

Sheet   plastination   of   human   brain   slices: Comparison between two staining methods. Jareonsuppapuch E, U  Tankittiwat,  P Padungchaichot. Department of Anatomy, Faculty of Medicine, Srinakharinwirot  University, Bangkok, Thailand.

Sheet plastinated brain slices are excellent teaching specimen in medical schools. Sheet plastinated brain slices utilizing resin produced in Thailand were prepared and stained. The aim of our study was to stain 2-6 mm thick sheet plastinated brain slices with Luxol fast blue and Astra blue staining methods. The result demonstrated that both stains clearly differentiate grey and white matter. The grey matter was white and the white matter was greenish blue in the Luxol fast blue staining method whereas the grey matter had blue color and the white matter showed white in the Astra blue staining method. In addition, the shrinkage of the two different stained sheet plastinated brain slices was insignificant.

Study of  brain  evolution  in  human  and  animals using the S/10 plastination method. Joghataei  MT; MH Asadi, A Mohammadzade, F Negahdar, G Jzadi­ Mobarakeh. Iran Medical University, Baqiyatallah University of Medical Science, Department of Anatomy, Tehran, Iran.

During the course of evolution, the area of brain that shows the most change is the cerebral hemispheres. The more recently evolved animals have a larger proportion of the brain taken up by the cerebral cortex. In the "higher" animals, the surface of the cerebral cortex becomes folded resulting in sulci and gyri. Material and Methods: This study used the S/10 plastination method on human, monkey, cat, cow, sheep, chicken, mouse, and fish brains. Results: Different neuroanatomical parts of the brains were compared. Also, sulci and gyri of mammal brains were compared. The results of the study are presented in a comprehensive table. Conclusion: Plastinated brains are available for demonstration of the evolutionary changes.

Computer image demonstrations combined with plastinated specimens: Applications for topographic lectures of head and neck clinical anatomy. Klepa  ek I, I Helekaf , J. Lexa2 Department of Anatomy, Kuwait University, Kuwait, 1Higher Professional School of Applied Art and Secondary School of Applied Art, Prague, Czech Republic, 2Biomedical Engineering Unit, Kuwait University, Kuwait.

The dissection of human cadaver materials may not provide all the topographical relations between human body structures. Because of our 2004 curricular schedule, it appears beneficial to establish an efficient method for lecture presentations to be oriented toward the postgraduate students. Lectures are followed up with clinically oriented anatomy. Here a combination of techniques are used which combine plastinated specimens with images of anatomical structures and computer applications. The dissection of the human material only does not always give correct topographical relations among human body structures. It appears that it is necessary to establish an efficient method for the lecture presentations oriented to the postgraduate students (due to our curriculum for 2004). We follow up the teaching process of  clinically oriented anatomy by combining pictures of the presented  topic,  plastinated  specimens  and  computer viewed applications. Materials and methods: Original pictures of the head and neck regions were made using the method of American retouch. The plastinated specimens (Kuwait and Prague anatomy collections; the Silicone S 10 procedure) and dissected organs served as master specimens. Using computer equipment  and digital audio/video system, the animated series of pictures and 3-D imagination were produces using software (ACDsee, Adobe-Photoshop, 3-D Anamaker). Results: Picture sets (based on curricular materials) were combined into four mutually "dissolving picture sets". These sets (15 pictures each) displayed structures from surface to depth. Photos of the plastinated specimens showing details of the selected topographic areas were incorporated into these sets. The following regions were illustrated : a) The face and orbit. b) The infratemporal fossa. c) The submandibular area. d) The neck triangles . Conclusion :  Desired relations among head and neck structures are presented using  picture sets, which are made as if seen by the physician eyes (if he is leaning towards patient). This is crucial to understand and remember mutual relationships, visual learning and an easy understanding of the external projection of the structures. Materials & Subjects & Methods: Original images of the head and neck regions were made utilizing American retouch technology . The silicone (SlO) plastinated specimens from the Kuwait and Prague anatomy collections and dissected organs served as the master models. An animated series of photos and 3-D imaging were produced using computer equipment, digital audio/video systems and production software (ACDsee , Adobe-Photoshop, 3-D Anamaker) . Results: Based on curriculum materials and needs, image sets were combined into mutually "dissolving image sets". Each set contained 15 images and were programmed to display anatomic structures from superficial to deep. Photos of the plastinated specimens showing details of the selected topographic areas were incorporated to the sets. The following regions were illustrated: 1. The face and orbit, 2. The infratemporal fossa, 3. The submandibular area, 4. The neck triangles. Conclusion: The programs are shown from the perspective of a physician leaning toward and viewing the patient. The sets show the relationships between head and neck structures. This methodology is an important way to reinforce the relationships of structures, visual learning and to understand the projection of deep structures under the surface.

The impact of plastination on the teaching of neuroanatomy. Lopez-Solerm M, 0 Roda-Murillo, A Roda-Murillo, I Ramirez-Ortiz. Department of Human Anatomy and Embryology, Medicine Faculty, University of Granada, Espana, Europe.

The development of plastination has allowed the introduction of new tools into the teaching of anatomy. Specimens obtained by  this method are dry, odorless, long-lasting and non-toxic . The present study compared the degree of satisfaction of students with the utilization of plastinated versus formalin preserved specimens for the study of  neuroanatomy.  Materials and methods: Analysis of variance was used to determine whether there was a relationship between the type of preservation technique (formalin  or plastination) as an independent variable and the degree of satisfaction as the dependent variable. Whether there was equality of means for the two techniques was investigated . Results : Equality of means was not observed . The independent variable (type of technique) significantly influenced the dependent variable (degree of satisfaction). The mean obtained for the plastination technique was higher than that for the formalin technique. Therefore , there was a greater degree of satisfaction with plastinated versus formalin preserved specimens. Conclusion: The plastination technique positively influences the degree of satisfaction of the student.

Plastination of a canine  encephalon  with hydrocephalus. Martin -Alguacil N, R Martin -Orti. Department of Anatomy and Comparative Pathologic Anatomy, School of Veterinary Medicine, Universidad Complutense de Madrid, Espana, Europe.

It is important to visualize the three-dimensional features of the ventricular system in order to understand how some pathological processes  such  as hydrocephalus can occur. Hydrocephalus is an increase in the volume of the cerebrospinal fluid, resulting in an extreme expansion of all the ventricles.  Brain prosections and brain slices were plastinated to study the ventricular system of the dog encephalon. Sections of a dog brain with hydrocephalus were plastinated as well. All specimens were plastinated with  silicone using the Biodur S 10 technique. In order to minimize shrinkage of the brain tissue, the  standard  procedure was slightly modified as recommended by Weiglein (2000). Prior to forced impregnation , the brains were immersed in a mixture of SIO/S3 at -20°C for three days. Good quality specimens were obtained after plastination of all tissues . A very good correlation between anatomical structures was  seen, both  before the specimens were fixed and after they were plastinated. Plastinated specimens of central nervous system  provide   the   student  with   a  useful   tool   to understand the dog ventricular system.

Anatomy of the mammary gland  of  the  goat  using SlO plastinated specimen. Martin -Alguacil N, R Martin-Orti, A Garcia, A Gomez, L Gallego. Department of Anatomy and Comparative Pathologic Anatomy, School of Veterinary Medicine,  Universidad Complutense de Madrid, Espana, Europe.

Understanding the anatomy of the mammary gland is important not only for veterinary students but also for practitioners while undertaking surgery  on that  region. The aim of this project was to provide a  plastinated model to study the anatomy of the mammary gland. The tissue was obtained from adult goats. To avoid  blood clots, the goats were heparinized  prior  to  euthanasia. The vascular system was washed by perfusion of saline solution and then the arteries and veins  were  injected with latex colored with red and blue pigments. The collecting sinuses were dilated due to the presence of milk. The tissue was frozen and cut in 2.5 mm sections. The slices were fixed in 5% formaldehyde for two weeks. The mammary  gland  sections were  dehydrated in increasing baths of acetone and plastinated following the standard  S 10  plastination  technique.  Nice specimens were obtained as a result. There was a good correlation between fresh and plastinated tissue.

Use of plastinated specimens to visualize the vascularization and inervation of the thorax. Martin-Orti R, N Martin-Alguacil. Department of Anatomy and Comparative Pathologic Anatomy, School of Veterinary Medicine, Universidad Complutense de Madrid, Espana, Europe.

Plastination is a great tool to use in teaching veterinary anatomy. It is also of a great value for student self­ directed learning. Students can freely access the prosection and visualize the different structures without spending a lot of time preparing a dissection. Herein we present a plastinated thorax from a foal that was dissected to study the thoraic vascular distribution and innervation as well. Special attention was given to show the thoracic autonomic nerves. Among these structures sympathetic trunk and ganglion, cervicothoracic ganglion, vagosympathetic trunk, recurrent laryngeal nerve and cardiac autonomic nerves were dissected and preserved. The thorax was plastinated using the S-10 technique with slight modifications. A very good quality specimen was obtained.

Room    temperature    degreasing    body    sections. Moreno M , H Abellan1, MD Ayala 1 C Sanchez1   Latorre 1Medicina, Universidad de Murcia, 1Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Murcia, Espana, Europe.

The main objective of this work was to compare two room temperature degreasing processes, a short one (3 weeks) and a long one (6 months). A whole fresh human body was frozen at -40C and transversely cut into sections of lcm thickness. Fixation was done by immersion in a 10% formalin solution and dehydration according to the regular system in cold acetone. Sections from head to cranial part of thoracic cavity were degreased in room temperature acetone for 3 weeks. The other sections were degreased in room temperature acetone for 6 months, (until acetone baths did not tum yellow). The short degreasing process had no shrinkage but the fat tissue had a bad appearance and the sections a bad smell. Sections from the long degreasing process had transparent odourless fat but more shrinkage than short time process.

Sheet plastination of the human midbrain slices demonstrating        dopaminergic       neurons. Padungchaichot P, E-0 Jareonsuppapuch, U Tankittiwat. Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.

Parkinson's disease 1s one of the maJor neurodegenerative disorders. The pathology of the disease is a specific degeneration of dopamine neurons in the midbrain. This was an applied study which utilized the sheet plastination technique for permanent preservation of immunohistochemically processed 1-2 mm thick-midbrain slices demonstrating dopaminergic neurons. Either DMSO or citrate buffer solution was applied to reverse the loss of antigenicity that occurs with some epitopes in formalin-fixed human brain. Primary antisera against tyrosine hydroxylase were used to demonstrate dopaminergic neurons in the midbrain. The results showed dark stained dopaminergic neurons in the immunohistochemically processed midbrain. In addition, the stained neurons could be permanently preserved by the  sheet plastination technique.

Green iguana plastination with the S 10 technique in Macedonia. Pendovski L, V /lieski, G Nikolovski1. Department of Anatomy, Faculty of Veterinary medicine-Skopje, Macedonia, 1Department of Internal Disease, Faculty of Veterinary Medicine-Skopje, Macedonia, Europe.

Green   iguanas   (iguana   iguana),   the   most   popular common lizard, has grow in popularity as a domestic pet. Preservation of the green iguana using the silicone plastination method is an essential process  for storing the specimen as a reference collection for a long time. This work describes the complete process  for plastination of an iguana preformed by  the standard  S 10 silicone technique, according to von Hagens. The green iguana's thick skin provides an opportunity to prevent the process of impregnation with silicone from penetrating the skin. Also the iguana's body was fixed in 3% formalin and in a bad body  position  for more than three years. Its body was enveloped by its tail and its head was positioned below and between  its forelimbs. One of the main goals was to acquire more correct anatomical position of the iguana, especially during the curing stage. The green iguana was dehydrated by the freeze substitution  method  using three acetone baths (100%) at -25°C over a four week period . The fourth acetone bath  was  at  room temperature for the period  of five days. When the acetone purity in the last acetone bath remained over 99.5%, dehydration and defatting were considered complete. Forced impregnation was  preformed,  after the iguana was submerged in a mixture of silicone Sl0/S3 (100:0 .5) in a freezer at -20 °C, by continuous pumping for three weeks. The vacuum was slowly increased to assure better penetration  of  polymer-mix and hence prevent shrinkage of the iguana. The impregnation process was considered  complete  when the bubbles ceased and the pressure was stabilized at 3 mbar. Finally, the iguana was cured by exposure to S6 gas cure. After 5 days, the surface of the iguana was cured, but to assure that the depths of the iguana cured, a tube with syringe was inserted in the abdomen of the iguana and S6 was conducted via this into abdomen for more than seven days. The iguana was perfectly preserved  without  noticeable  shrinkage.  The penetration of silicone trough iguana skin was very successful in spite of its thickness.  Compared  with other plastinated specimens, there wasn't  a  big difference considering the skin thickness. During the curing stage, the iguana shape was reformed even after being fixed for over three years in an inappropriate position. The plastinated iguana looked  very  natural and can be easily carried and manipulated with less risk of damage.

Branches of the facial artery of the perioral region: Latex injection study. Pinar Y, 0 Bilge, F Govsa. Ege University Faculty of Medicine, Department of Anatomy, Ege University, Izmir , Turkey.

There are numerous techniques for reconstruction  of lip defects. The use of flaps requires a detailed knowledge of local vasculature . New flaps for the region  around the mouth can be devised if the surgeon is aware of the distribution of the perioral arterial branches. Materials and methods: Vessels supplying blood to the perioral region were examined by dissecting 25 cadavers  that were fixed with a 10% formaldehyde  solution.  Red latex was injected into the conunon carotid arteries before dissection. The size and  distribution  of  the vessels were investigated and the primary supplying vessels were identified . Results: The facial artery (FA) was symmetrical in 17 of the 25 heads (68%). It terminated as an angular facial vessel in 11 (22%), a nasal facial vessel in 30 (60%), a superior alar vessel in 6 (12%) and a superior labial vessel in 2 facial halves (4 %). It terminated as a hypoplastic type of FA in 1 facial half (2%). The average external  diameter of the superior labial artery at its origin was 1.6 mm. The beginning point of the superior labial artery (SLA) was above the angle of mouth in 34 out of 47 specimens (72.3%), and at the angle of mouth in 13 out of 47 specimens (27.7%). Three of the specimens were not evaluated because in 2 of these 3 the SLA was the continuation of LA and the other was  a  hypoplastic type. The SLA supplied the columellar branches in all samples except for the hypoplastic type (49 specimens). Columellar branches were classified according to their number or type. In 5 specimens (10 %), there  was no inferior labial artery (ILA). In the other specimens, the bifurcation point of the ILA varied between  the comer of the mouth and the inferior margin of the mandible. ILA arises from the FA above the angle of mouth in 4 (8.8%) specimens, under  the  angle of mouth  in  9 (20 %) specimens and, at the angle of mouth in 30 (66.6%) specimens . Conclusion : We suggest that knowledge of the artery's location with respect to easily  identifiable landmarks will help to avoid complications.

The experience of setting up  a  plastination laboratory. Reina-De-La Torre F, A.  Rodriguez­ Baeza, JM Domenech-Mateu, M Querol-Jimenez, I Delgado-Calvarro. Department of Morphological Sciences (Anatomy and Embryology Unit), Faculty of Medicine, Autonomous University of Barcelona, Barcelona , Espana, Europe .

Contrary to intuition, to perform the basic techniques of plastination does not require a significant economical investment. The technical equipment can be  obtained with a limited budget . The present work reports the recent experience in the design and setting up of a laboratory to plastinate anatomical  preparations  using the   standard   S-10  silicone   technique.   The  different ways used to finance the cost of the facility and the technical equipment will be explained . Materials and methods : The plastination laboratory was designed in accordance with the  International  Society  of Plastination guidelines  and  with  the  technical  support of the Plastination Laboratory of the University of Murcia. The financial resources came from the Faculty of Medicine , the Anatomy and Embryology budget or from external grants for increasing safety in research laboratories. The design of the plastination  laboratory was  carried  out  in  order  to  start  up  the  standard  S10 silicone technique . Results : The laboratory occupies a total surface of 12.5 m2 . The laboratory has a forced extraction   system  for  acetone  vapors.  The   financial resources allocated to the plastination laboratory were 10,418 euros. These resources were used as follows : 21.85% for the facility, 61.75% for technical equipment and 16.40% for laboratory materials. Conclusion : The setting up of a plastination laboratory does not need to be a high economical effort. On the contrary, the advantages of the technique justify the investment. It reduces the exposure to toxic fumes, improves the preservation of teaching preparations and promotes techniques in self-directed learning of anatomy. Preliminary   results   with   the   S10   standard technique:

An  experience  of  the   future   in   the Faculty  of  Medicine  of  the  Autonomous  University of Barcelona. Reina-De-La Torre F, A. Rodriguez­ Baeza, JM Domenech-Mateu, M Querol-Jimenez, I Delgado-Calvarro. Department of Morphological Sciences (Anatomy and Embryology Unit), Faculty of Medicine, Autonomous University of Barcelona, Barcelona, Espana, Europe.

According to the collection of technical leaflets for plastination from Heidelberg, the easiest and cheapest plastination methodology is the standard S10 silicone technique . With the technical support and collaboration of the plastination laboratory of the University of Murcia, a plastination experience was initiated in the Faculty of Medicine. The two main objectives were to initiate the elaboration of an anatomical collection of plastinated specimens in order to promote self-learning techniques in human anatomy and to reduce exposure to carcinogenic  toxic  fumes.  The  present  work   shows the preliminary results. Materials and methods: Specimens of various anatomical regions or systems (central nervous system, heart, hand, topographical slices, facial region and bones) from human cadavers previously fixed in Cambridge solution were processed according to the standard S10  silicone  technique. Results :   The   obtained   plastinated    specimens   were optimum with regard to preservation , color and durability. The processing time ranged from 1.5 to 4 months. Specimens with the most difficulties for dehydration and impregnation were the facial and hand regions. Vascular Injection with colored latex complimented the process. The use of plastinated specimens for practical activities in the medicine curriculum has been received by the students enthusiastically. Specimens can be manipulated without disruption, they are odorless and they may be used outside the programmed sessions. Conclusions: The standard S 10 silicone technique is a very standardized method, not difficult from a  technical  point  of  view, and offers a great range of possibilities to improve the teaching of anatomy: It reduces the exposure  of teachers and students during practical activities to carcinogenic toxic fumes, It better preserves teaching specimens, It renders access to such specimens by students, and It promotes techniques in self-directed learning. There is no doubt that the technique will have an important role in the future changes of higher education curricula.

Plastination  in   teaching   neuroanatomy.   Roda­ Murillo 0, Lopez -Solerm M, A Roda-Murillo, I Ramirez-Ortiz. Department of Human Anatomy and Embriology, Medicine Faculty, University of Granada, Espana, Europe.

The difficulty of obtaining human brains for practical classes in Human Neuroanatomy  and  the  fragility  of this material has led to the development of new preservation techniques to obtain more resilient human brain specimens that are faithful to reality and suitable for handling by students. The  present  study compared the value of plastinated versus formalin preserved specimens in neuroanatomy teaching. Materials and methods : A questionnaire was administered to students of neuroanatomy on the use of formalin preserved and plastinated specimens. It asked: Which technique better preserved and allowed longer use of the specimen, Which specimens were more faithful to reality and better to handle, and Which technique was preferred by the students. The results were subjected to statistical analysis. Results: 90.8% of the students considered plastination preserved specimens better when compared with formalin preservation. 93.3% believed plastinated specimens to be faithful to reality. 40.8% thought they were more realistic than formalin preserved specimens, 45.4% thought they were equally realistic, and  only 13% described the plastinated specimens as  less realistic. All of the students believed that the handling was  better  with  plastinated  versus  formalin  preserved specimens. 93.6% preferred plastinated specimens for the study of neuroanatomy and 6 .4% preferred formalin preserved specimens. Conclusion: Plastination is a very valuable tool for teaching of neuroanatomy.

Plastination and mimimally invasive surgery. Sachez Margallo FM, J Uson, F Soria, F Gil1 R Latorre 1.0 Lopez        -Albors1,  M Sarasa 1 S  Climent1. Minimally invasive surgery Center, Caceres, Spain, Europe. Medicina y Cirugia Animal, Facultad de Veterinaria, Universidad de Murcia,  Espana. 1Anatomia y Embriologia, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, Murcia, Espana, Europe.

Plastination is a unique technique of  tissue preservation. Plastinated  organs  are useful as didactic resources when teaching minimally invasive surgery. The morphological and physical characteristics of plastinated specimens are excellent  to  study topographic and clinical anatomy. Specially designed plastinated organs aid the teaching-learning process of minimally invasive surgical techniques, as they allow training of endoscopic techniques and skills. These techniques will be shown in a DVD presented. Digestive system: Whole or  partial  gastrointestinal tracts of the dog were specially designed for exercises in digestive endoscopic explorations (esophagogastroscopy, duodenoscopy and colonoscopy) and plastinated. Visceral topography and the endoluminal aspects of plastinated organs are real and their flexibility allows exploration with a regular digestive endoscopic. Cardiopulmonary System: Plastinated cardiopulmonary blocks and isolated lungs of dogs, were designed to practice respiratory endoscopic explorations (bronchoscopy, cytology and tracheal suction and bronchial flushing). Isolated and plastinated hearts were used as a vascular model for endoluminal and interventional radiology techniques. Whole cardiopulmonary casts, bronchial tree casts and colored silicone pulmonary vessels casts of pigs and sheep were used as didactic tools for thoracoscopic surgery. Reproductive System: Plastinated female genital tracts of dogs and sheep and male genital tracts of dogs and pigs, suitable for gynecological laparoscopy    surgery,    were   prepared   and   used  as anatomical didactic models. Urinary  System: Kidneys, ureters, urinary  bladder and urethra in abdominal and pelvic cavities of the dog, pig and sheep were vascular injected and dissected. These plastinated specimens were    used    for   teaching    experimental    urological laparoscopy.

Embalming technique and dissection procedure of human bodies  at  the  University  of  Antwerp, Belgium. Van Toor I, H Bortier. Human Anatomy and Embryology, Department of Medicine, University of Antwerp, Antwerpen, Belgium, Europe.

Personal health of the technician and the students as well as environmental considerations led to the adaptation of this embalming technique and the dissection procedure for human bodies. It is known that formalin and phenol vapours cause irritation of human airways and cause environmental concerns. Second, there is only one technician at the institution to embalm all of the human bodies. Third, the educational developers of the new curriculum asked whether it would be possible to work  in smaller groups. For all these reasons the embalming technique and the dissection procedure were adapted as follows. Materials and methods: An ethanol solution is injected as the embalming fluid via a catheter in the A femoralis. Second, in order to shorten the embalming period, a pump is fitted to the catheter. Third, during the dissection course, six first year medical students who are coached by at least one medical student dissect one body. Results:  According to the comments  of the technician, the students, the environmental adviser and the medical coaches, there is less irritation of the airways, the human bodies smell less and the department has taken care of the environmental concerns. Second, the technician is happy that embalming a human body only takes three to four hours. Third, the medical students and the curriculum developers are pleased that there is more space to work and that they are coached better. Conclusions The embalming technique and the dissection  procedure of human bodies at the University of Antwerp were successfully adapted to the problems and the questions put forward.

 

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