Low viscosity silicone with less shrinkage for brain slices.

Plastination consists of replacing lipid and water with a curable polymer. This technique has numerous advantages, of which the production of non-toxic, inert, highly durable, dry, and easy maintenance and storage specimens stand out. Like all anatomical techniques, plastination also has disadvantages, and one of them is tissue shrinkage. The feasibility of using low viscosity domestic silicone (0,1Pa.s at 20°C) to plastinate brain slices was examined. Twenty humans, 10 millimeters (mm) brain slices were impregnated, ten slices each with two polymers [10 with domestic low viscosity polymer - P1 and 10 slices with Biodur® (0,45-0,6Pa.s at 20°C) S10]. Shrinkage was accessed by volume and area measurements. Volume shrinkage was significantly less in the slices impregnated with low viscosity domestic polymer, demonstrating the feasibility to plastinate brain slices with domestic low viscosity silicone polymer.

Resin-embedded anatomical cross-sections as a teaching adjunct for medical curricula: is this technique an alternative to potting and plastination?

With an ever-expanding use of cross-sectional imaging for diagnostic and therapeutic purposes, there has also been an increase in the need for exposure to such radiological and anatomical views at the undergraduate and postgraduate level to allow for early familiarisation with the relevant anatomy. Cadaveric cross-sections offer an excellent link between the two-dimensional radiological images and the three-dimensional anatomical structures. For such cross-sections to be useful and informative within educational settings, they need to be: (i) safe for students and trainees to handle and (ii) robust enough to withstand repeated handling; as well as (iii) displaying anatomy clearly and accurately. There are various ways in which cross-sections can be prepared and presented; plastinated, potted, vacuum-sealed or unmounted. Each of these approaches has advantages and disadvantages in terms of technical complexity, cost and quality. As an alternative to the above methods and their limitations, we propose the presentation of cadaveric cross-sections in a transparent polyester resin. This technique has been used extensively in craft and artistic industries, yet it is not publicised in anatomy teaching settings. The sections were layered in polyester resin contained within a mould. The set resin required finishing by sanding and polishing. The final cross-sections were safe to handle, durable and maintained excellent anatomical relationships of the contained structures. The transparency of the set resin was water-clear and did not obstruct the visibility of the anatomy. The cost of the process was found to be significantly lower, requiring less infrastructure when compared with alternative methods. The following trivial technical difficulties were noted during the resin-embedding process: trapped air causing organs to float; retained water in the anatomical specimens creating bubbles and discoloration; and microbubbles emerging from the solution affecting the finished surface. However, solutions to these minor limitations have been discussed within the paper with the aim of future proofing this technique. The sections have been used in undergraduate medical teaching for 4 years and they have shown no signs of degradation or discoloration. We believe that this method is a viable and cost-effective alternative to other approaches of displaying cross-sectional cadaveric material and will help students and trainees bridge the gap between the traditional three-dimensional anatomy and two-dimensional images.

Preservation of macroparasite species via classic plastination: an evaluation.

Plastination is a preservation method for biological specimens, with important advantages over classic conservation techniques with formaldehyde or alcohol. Plastinated specimens are dry, odourless, and free of carcinogenic and toxic solutions. There are only few references about the plastination of parasites. Moreover, there is no information on the effect of plastination on the morphology and morphometry of these animals. The aim of this study was to define a plastination protocol to preserve various species of parasites, namely the nematodes Parascaris equorum (Goeze, 1782); Ascaris suum Goeze, 1782 and Dirofilaria immitis (Leidy, 1856); the acanthecephalan Macracanthorhynchus hirudinaceus (Pallas, 1781); the trematodes Fasciola hepatica Linnaeus, 1758 and Dicrocoelium dendriticum (Rudolphi, 1819) and the tapeworm Taenia sp. in the best morphological and morphometric conditions. Results showed that some individuals suffered collapse (P. equorum, A. suum, and D. dendriticum). However, other parasites presented good results with almost no change after plastination (D. immitis, M. hirudinaceus and F. hepatica). In conclusion, conventional plastination allowed anatomical preservation of all helminths tested, but modifications to the protocol are needed to prevent collapse.

High field magnetic resonance imaging anatomy of feline carpal ligaments is comparable to plastinated specimen anatomy.

Feline carpal ligament injuries are often diagnosed indirectly using palpation and stress radiography to detect whether there is instability and widening of joint spaces. There are currently no reports describing normal feline carpal ligament anatomy and the magnetic resonance imaging (MRI) appearance of the carpal ligaments. The objective of this prospective, anatomic study was to describe normal feline carpal ligament anatomy using gross plastinated specimens and MRI. We hypothesized that MRI could be used to identify the carpal ligaments as previously described in the dog, and to identify species specific variations in the cat. The study was conducted using feline cadaver antebrachii that were radiographed prior to study inclusion. Three limbs were selected for MRI to confirm repeatability of anatomy between cats. The proton density weighted pulse sequence was used and images were acquired in transverse, dorsal, and sagittal planes. Following MRI, the limbs were plastinated and a collagen stain was used to aid in identification of carpal ligament anatomy. These limbs were sliced in sagittal section, and a further six paired limbs were included in the study and sliced in transverse and dorsal planes. Anatomic structures were initially described using MRI and then subjectively compared with gross plastinated specimens. Readers considered the transverse MRI plane to be most useful for visualizing the majority of the carpal ligaments. Findings indicated that MRI anatomy of the carpal ligaments was comparable to plastinated anatomy; therefore MRI appears to be a beneficial imaging modality for exploration of feline carpal pathology.

Through the fingers: Use of plastinated anatomical specimens for visually impaired students.

The plastination technique produces non-toxic human tissues, ensuring their safe handling in educational settings. This investigation aimed to understand if visually impaired students profit from the use of plastinated anatomical specimens in learning the anatomy of the nervous system. For this purpose, their learning performance was compared to sighted and blindfolded students recruited from three primary schools in Fortaleza city, in the state of Ceará. Initially, a questionnaire was applied before carrying out the pedagogical practice, followed by an anatomy lecture with practical components with the use of plastinated anatomical specimens and synthetic anatomical models of the nervous system. After these steps, the students answered the questionnaire previously applied. Our results showed that the tactile perception of the visually impaired participants was significantly more developed compared to sighted (p < 0.001) and the blindfolded (p < 0.0001) students. The average of correct answers in the reapplied questionnaire was higher in the groups that used plastinated specimens (p < 0.05). In conclusion, the use of plastinated specimens has proven to be an effective tool in promoting a better understanding of anatomical structures, mainly for students with or without visual impairments, making it a valuable asset in anatomy teaching.

Efficacy of plastinated specimens in anatomy education: A systematic review and meta-analysis.

Plastination, a permanent preservation method for human tissues and organs, is increasingly being used in anatomy education. However, there is a paucity of systematic reviews and meta-analyses summarizing the educational efficacy of plastinated specimens. This meta-analysis compared the assessment scores of students exposed to plastinated specimens against those exposed to other common instructional methods. A systematic search was conducted through four databases, from 2000 to July 2022. Titles and abstracts of the retrieved records were screened according to predetermined eligibility criteria. Of the 159 records screened, 18 were subjected to full-text review. Among the 18 studies, five articles reported post-intervention test scores for intervention (plastinated) and control (other modalities) groups. Studies were subjected to GRADE quality assessment, and four studies with moderate to high ratings were included for meta-analysis. Students' perceptions (n = 15 studies) were qualitatively analyzed using an inductive narrative analysis. No significant effect was detected between the intervention (n = 417) and control groups (n = 422) (standardized mean difference = 0.08; 95% CI [-0.36, 0.52]; p = 0.73). Four themes emerged from students' perceptions: ease of use, motivation to study, spatial understanding, and learning preference. Overall, student performance outcomes comparing the use of plastinated specimens versus other instructional modalities are very limited. This meta-analysis suggests that knowledge gained from plastinated specimens is comparable to learning achieved through other modalities; though this outcome should be interpreted with caution as there is currently insufficient evidence for definitive conclusions.

Morphometric and histopathological evaluation of modified Elnady's plastinated tissue compared to non-plastinated tissue: Highlighting its relevance for teaching and research.

The present study aims to evaluate the morphometric and histopathological properties of Modified Elnady's plastinated tissue after a period compared to non-plastinated tissue. The plastination technique is utilized in research and teaching due to the potential health risks associated with prolonged exposure to formalin. The tissues and organs are permanently dried during plastination and can be used for further anatomical, histopathological and surgical educational purposes. This method involves drying tissue and allowing synthetic materials like glycerin to permeate it. The study compared non-plastinated and plastinated tissue post-plastination to determine if structural alterations differed from those linked to plastination. The study examined the histopathological examination of dogs' skin, muscles, liver, lung, and intestine using formalin-fixed organs for paraffin embedding and previously plastinated organs for a plastinated group. The study examined non-plastinated and plastinated tissues, their histological composition and biometric parameters revealing typical structures in the non-plastinated group. Plasmodiumted tissues exhibited a compacted appearance, volume changes, nuclear clarity, and cytoplasmic hypereosinophilia, with statistical differences between the two groups. The study reveals that plastinated tissues, after 5 years of plastination, maintain their histological architecture well, with some exceptions. Plastinated tissues can be utilized in future microscopic and immunological studies and will be beneficial for teaching and research.

Is domestic polyester suitable for plastination of thin brain slices?

Plastination is a technique used to preserve biological tissues while retaining most of their original appearance. In the technique, developed by Dr. Gunther von Hagens in 1977, specimens were impregnated with a polymer, such as silicone, epoxy, or polyester. Considered the most suitable material for brain plastination, polyester has a wide application in teaching and research compared with imaging techniques. The materials for plastination are usually imported from Germany and more expensive than domestic products. If domestic polymers were to enter the market it would favor the expansion of plastination in Brazil. Hence, this study evaluated the feasibility of using domestic polyesters to replace the usual Biodur® (P40) in plastination of brain slices. For this evaluation, 2-mm-thick sections of bovine brains were prepared and plastinated with domestic polyester. Slices were compared before impregnation and after curing using standardized photographs taken after dehydration and after curing. Plastination followed the standard protocol: fixation, dehydration, forced impregnation, and curing. Fifteen brain slices were plastinated with each polyester (P40, P18, and C1-3). There was no significant difference in the percent shrinkage between groups after plastination of P18 and P40, but the curing time of Cristalan© polymer was too short for impregnation. Therefore, no initiator was used for C polymers impregnation. Thus, domestic polyester P18 was a viable option for the process.

An improved plastination method for strengthening bamboo culms, without compromising biodegradability.

Biomaterials are increasingly being designed and adapted to a wide range of structural applications, owing to their superior mechanical property-to-weight ratios, low cost, biodegradability, and CO2 capture. Bamboo, specifically, has an interesting anatomy with long tube-like vessels present in its microstructure, which can be exploited to improve its mechanical properties for structural applications. By filling these vessels with a resin, e.g. an applied external loading would be better distributed in the structure. One recent method of impregnating the bamboo is plastination, which was originally developed for preserving human remains. However, the original plastination process was found to be slow for bamboo impregnation application, while being also rather complicated/methodical for industrial adaptation. Accordingly, in this study, an improved plastination method was developed that is 40% faster and simpler than the original method. It also resulted in a 400% increase in open-vessel impregnation, as revealed by Micro-X-ray Computed Tomography imaging. The improved method involves three steps: acetone dehydration at room temperature, forced polymer impregnation with a single pressure drop to - 23 inHg, and polymer curing at 130 °C for 20 min. Bamboo plastinated using the new method was 60% stronger flexurally, while maintaining the same modulus of elasticity, as compared to the virgin bamboo. Most critically, it also maintained its biodegradability from cellulolytic enzymes after plastination, as measured by a respirometric technique. Fourier transform infrared-attenuated total reflection, and thermogravimetric analyses were conducted and showed that the plastinated bamboo's functional groups were not altered significantly during the process, possibly explaining the biodegradability. Finally, using cone calorimetry, plastinated bamboo showed a faster ignition time, due to the addition of silicone, but a lower carbon monoxide yield. These results are deemed as a promising step forward for further improvement and application of this highly abundant natural fiber in engineering structures.

Influence of Plastination on Ultrasound Transmission Through the Human Skull.

Development, optimization and validation of transcranial ultrasound methods require the use of fresh human or animal skulls. However, to avoid fresh skull degradation over time, fixation methods are required for conservation, such as formaldehyde buffer solution. This method allows for conservation of the skull properties over a relatively long period, but requires specific conditioning (de-gassing) and storage conditions, such that its practical use is limited. Plastination appears to be a unique solution for the preservation and transportation of body parts without constraints. However, the influence of this conservation process has yet to be characterized with respect to ultrasound transmission to verify that the acoustic and mechanical properties of the skulls are not altered by the plastination process. The objective of the study described here was to quantify the effect of plastination on ultrasound transmission through the temporal and parietal areas of the human skull between 200 kHz and 2 MHz. To achieve this, transmission measurements were performed on three different skulls and four areas before and after plastination. It was found that the plastination process results in a transmission loss of 5 dB. Moreover, results indicate that the plastination process does not induce any phase shift in the transmitted signal, validating the proper use of plastinated skulls for in vitro measurements and development of new transcranial ultrasound methods.

Plastination with low viscosity silicone: strategy for less tissue shrinkage.

Plastination is an anatomical technique for preserving biological tissues based on the principle of replacing body fluids with a curable polymer. An inconvenient aspect of this technique is the tissue shrinkage it causes; several studies seek ways to reduce or avoid this shrinkage. Additionally, there are no studies in the literature that quantitatively evaluate the use of low viscosity silicones in plastination having shrinkage of tissue as a parameter. Therefore, this study aimed to evaluate the use of Silicones S10 (Biodur) and P1 (Polisil) in the plastination of different types of biological tissues of a sliced human body, having as a parameter the tissue shrinkage caused in the forced impregnation stage. Human cardiac, pulmonary, splenic, renal, hepatic, muscular, and bone tissues were analyzed. For such purpose, a male human body was used, sliced in 13-15-mm-thick pieces, having as a parameter the before and the after plastination with the different silicones. The standard protocol of the plastination of the slices was followed: dehydration, forced impregnation, and curation. Half of the pieces obtained were plastinated with silicone P1 (group P1) and the other half with S10 (group S10). All tissues and anatomical segments analyzed in this study showed less or equal shrinkage when plastination of the control group (S10) was compared with that of the P1 group. Therefore, we concluded that the lower viscosity silicone promoted less tissue shrinkage, making it a viable alternative to the reference.

Colorimetric evaluation of cross-sectional silicone plastination of the Total head region of sheep and Deplastination of the histological sections of brain tissue.

The aim of the study is to protect and preserve the cross-sectional diagnostic characteristics of the anatomy samples by using silicone plastination method, to examine them both macroscopically and microscopically, and to use them as an educational material. After the dissection procedures of 10 total sheep heads obtained from the slaughterhouse were completed, they were freshly frozen and sliced to prepare cross-sectional samples. Then, statistical analysis was performed after the colorimetric measurements. For microscopic examination, 30 brain samples were divided into three groups (Fresh-F, plastination-P, plastination/deplastination-P/D). Of the total brain samples, 20 were subject to routine plastination protocol. After the plastination/deplastination procedure, the changes occurring in cerebral histology were compared. In terms of tissue preservation, the effect of plastination and deplastination was examined using a light microscope. Plastinates subject to silicone plastination under room temperature were very similar to their natural appearance, and it was observed that they preserved their morphological features. Colour changes in the tissues were statistically evaluated. Volumetric shrinkages were observed as qualitative, especially in the brain. As a result of the evaluation done, it was seen that deplastination with toluene is not possible for the brain tissues. In addition, it was not possible to take cross sections of the plastinated tissues that were not deplastinated. On the contrary, findings regarding that deplastination with 5% sodium methoxide dissolved in methanol can allow microscopic examination in long-term preserved plastinated brain tissues were obtained.

Fibrous configuration of the fascia iliaca compartment: An epoxy sheet plastination and confocal microscopy study.

BACKGROUND AND OBJECTIVES: The underlying anatomical mechanism of the ultrasound-guided fascia iliaca compartment (FIC) block for anaesthesia and analgesia in the lower limb has not been illuminated and numerous variations were attempted to achieve an optimal needle placement. This study aimed to define the fibrous configuration of the FIC. METHODS: A total of 46 adult cadavers were studied using dissection, latex injection, epoxy sheet plastination and confocal microscopy. RESULTS: (1) The fascia iliaca originated from the peripheral fascicular aponeurotic sheet of the iliopsoas. (2) The FIC was a funnel-shaped adipose space between the fascia iliaca and the epimysium of the iliopsoas, had a superior and an inferior opening and contained the femoral and lateral femoral cutaneous nerves but not obturator nerve. (3) The estimated volume of the FIC in the cadavers was about 23 mls, of which about one third was below the level of the anterior superior iliac spine. CONCLUSIONS: This study revealed that the fascia iliaca was aponeurotic and may be less permeable for the local anesthetics. CONCLUSIONS: The FIC contained only the femoral and lateral femoral cutaneous nerves and communicated with the extraperitoneal space and femoral triangle adipose space via its superior and inferior opening, respectively.

Development of an ultrathin sheet plastination technique in rat humeral joints with osteoarthritis induced by monosodium iodoacetate for neovascularization study.

Injection with monosodium iodoacetate (MIA) is widely used to produce osteoarthritis (OA). Ultrathin sheet plastination has been used to study the morphology of structures, with strong application in anatomical education and research. Our aim was to carry out, for the first time, ultrathin sheet plastination of rat humeral joints to observe the neovascularization provoked by OA. We injected 0.1 mL of MIA into the left humeral joints of ten Sprague-Dawley rats. The right shoulders of the same rats were used as control. Sixteen weeks after the injection, the animals were euthanized and were given an immediate red epoxy resin injection through the thoracic aorta. The samples were fixed in 10% formalin, prior to the plastination process, without decalcification. Samples were dehydrated with acetone (100%) at - 25 °C, for 10 days. Later, for degreasing, samples were immersed in methylene chloride at room temperature during 1 week. Forced impregnation was performed inside a stove within a vacuum chamber. The plastinated blocks obtained were cut with a slow velocity diamond blade saw. Slices were placed in curing chambers to achieve curing and final tissue transparentation. 230 µm thickness slices were obtained. The slices were analyzed under magnifying glass and microscope, achieving visualization of OA neovascularization. The cartilage affected by OA loses its ability to remain avascular, and blood vessels invade it from the subchondral bone to the calcified and uncalcified cartilage. Ultra-thin sheet plastination is useful to observe articular cartilage neovascularization, caused by OA induced with MIA in humeral rat joint.

Room temperature/Corcoran/Dow Corning™-Silicone plastination process.

For 20 years, the cold temperature/S10/von Hagens' plastination technique was used to preserve biological specimens without challenge. It became the "gold standard" for preservation of beautiful, dry biological specimens. Near the end of the 21st century, a group from the University of Michigan and environs and Dow Corning™, USA, combined silicone ingredients, similar to the von Hagens' plastination products, however in a different sequence. The new polymer (Cor-tech) was combined with the cross-linker to design the "impregnation mix" which would invade the cellular structure of the specimen and yet was stable at room temperature. Later, curing would be by application of the catalyst onto the impregnated specimen. This unique sequencing of products would become the "Room temperature/Dow Corning™/Corcoran-Silicone plastination technique." The results of this room temperature technique provided similar plastinates, beautiful and practical for demonstration, containing no toxic chemical residues and forever preserved. As the name implies, impregnation of this silicone mix could be done at room temperature, without having to be kept cold. Both processes (cold and room temperature) required the same four basic steps for plastination. As well, both processes used similar basic polymers and additives to produce plastinates. However, they were combined in a different sequence. Cold temperature combines polymer and catalyst/chain extender, which is not stable and therefore must be kept colder than -15°C, while room temperature combines polymer with cross-linker which is stable, and likely forever.

Cold temperature/Biodur® /S10/von Hagens'-Silicone plastination technique.

Plastination is a late 20th century preservation methodology which replaces tissue fluid within a specimen with a curable polymer, such as silicone. Plastination yields superb, beautiful, well-preserved specimens each with their own unique qualities. Silicone polymer is used around the world to preserve macroscopic cadavers or portions/organs thereof. Plastination was conceived by Dr. Gunther von Hagens, Universität Heidelberg, Heidelberg, Germany prior to 1977. Silicone polymer was the primary polymer which emerged initially for plastination. The Biodur® line of silicone polymer and additives was chosen and manufactured because it has consistently produced the best plastinates since the inception of plastination. Since the discovery of silicone, generic and similar silicone polymers are known and used around the World by many industries and used in numerous products. The plastination process has four steps: Specimen preparation, Specimen dehydration and degreasing, Vacuum-forced impregnation of specimens and Specimen hardening.

Plastination of larger and massive specimens-With Silicone.

In 1977, plastination was unveiled, which replaced tissue fluid with a curable polymer. Today preservation via plastination of various animal and plant tissues, organs, and whole bodies is an extremely useful technique to display such and help educate vast arrays of both allied science students and the lay public across the planet. The diversity of applications of plastination techniques seems to be without limits. In fact, the only real limitation to plastination is one's imagination! The size of plastinates during the early years of plastination was comparatively small and dictated primarily by the size of the available plastination kettle/chamber, 35 L Heidelberg plastination kettle (49 cm H × 34.5 cm diam.). In the 1990s larger chambers were designed and slowly became available:150-210 cm (long) × 65-80 cm (wide) and 83-92 cm (high). Today a few large vacuum chambers are in service which will accommodate whole bodies of man and domestic or exotic animals. Today, at least two gigantic chambers are available to impregnate massive specimens. These are 3.5 m × 2 m × 1.5 m (Dalian) and 4 m × 3 m × 2.2 m (Guben). Also, the need for larger quantities of acetone and impregnation mix, not to mention the great increase in specimen preparation time, makes this a major investment. The "cold temperature process" is used to impregnate these massive creations. The room temperature technique could be used. The same four plastination steps are necessary for larger and massive specimens. Besides their tremendous size, the slippery silicone polymer is a reckoning force.

Multiple polymer plastination: Combining different types of polymers in teaching and exhibition plastinates.

Vacuum forced tissue impregnation is the signature step of the plastination process. It requires polymers with a low vapour pressure, low viscosity and a long pot life. Plastination polymers are a compromise between these mandatory requirements on the one hand and various secondary demands such as specimen stability, resistance to UV light and defined light refraction index on the other hand. Combining different polymers in one plastinate instead of using one plastination polymer alone can result in improved specimens for exhibitions and teaching including hands-on use for students. The aim of this study was to assess the range of possible sheet plastinate modifications and how the resulting multiple polymer plastinates can fulfil the secondary requirements of user-friendly plastinates. Adding sub-steps of tissue impregnation and processing to the standard plastination protocol allows combining different polymer properties including the use of substances which are not suitable for conventional plastination as such but have better properties than plastination polymers. Advantages like resistance to UV light and mechanical stability can be combined and characteristic disadvantages of plastination polymers can be avoided. Acrylic protection layers (APL) offer a complete protection of the specimen in combination with advanced presentation possibilities and the option of completely refurbishing valuable specimens. Hybrid sheet plastinates provide lower preparation cost and polymer-tissue interactions for an improved visualization of fat, nerves and brain tissue. Selective impregnation is a promising approach for the clearer differentiation of various structures and tissue types.

E12 technique: Conventional epoxy resin sheet plastination.

Epoxy plastination techniques were developed to obtain thin transparent body slices with high anatomical detail. This is facilitated because the plastinated tissue is transparent and the topography of the anatomical structures well preserved. For this reason, thin epoxy slices are currently used for research purposes in both macroscopic and microscopic studies. The protocol for the conventional epoxy technique (E12) follows the main steps of plastination-specimen preparation, dehydration, impregnation and curing/casting. Preparation begins with selection of the specimen, followed by freezing and slicing. Either fresh or fixed (embalmed) tissue is suitable for epoxy plastination, while slice thickness is kept between 1.5 and 3 mm. Impregnation mixture is made of epoxy E12 resin plus E1 hardener (100 ppw; 28 ppw). This mixture is reactive and temperature sensitive, and for this reason, total impregnation time under vacuum at room laboratory temperature should not last for more than 20-24 hr. Casting of impregnated slices is done in either flat chambers or by the so-called sandwich method in either fresh mixture or the one used for impregnation. Curing is completed at 40°C to allow a complete polymerization of the epoxy-mixture. After curing, slices can be photographed, scanned or used for anatomical study under screen negatoscope, magnification glass or fluorescent microscope. Based on epoxy sheet plastination, many anatomical papers have recent observations of and/or clarification of anatomical concepts in different areas of medical expertice.

P40 polyester sheet plastination technique for brain and body slices: The vertical and horizontal flat chamber methods.

The P40 technique produces high-quality brain and body slices and is the user-friendliest of the polyester techniques. The P40 polyester technique follows the same classical steps for plastination. That is, preparation of the specimen, fixation (optional), dehydration by freeze substitution, forced impregnation and curing. Two methods used to prepare two different types of specimens, that is, brain slices and body slices are described. Each method has its own characteristics depending on the specimen type used. Brain slices were used to illustrate the vertical small chamber method while the body slices were used to illustrate the horizontal large chamber method. The brain slices obtained using P40 are of very good quality presenting good contrast between grey and white matter. The body slices are also of very good quality. The physical appearance of these slices makes them an exceptional instrument for diagnostic imaging and anatomical correlation. Body slices prepared with P40 retain the natural colour of the tissue and preserve the anatomical relationships.