Body Slices Research Articles

A Segmentation Method of Serialized Human Body Slices Based on Matting Strategy and Skeleton Extraction.

In this paper, a semiautomatic image segmentation method for the serialized body slices of the Visible Human Project (VHP) is proposed. In our method, we first verified the effectiveness of the shared matting method for the VHP slices and utilized it to segment a single image. Then, to meet the need for the automatic segmentation of serialized slice images, a method based on the parallel refinement method and flood-fill method was designed. The ROI (region of interest) image of the next slice can be extracted by using the skeleton image of the ROI in the current slice. Utilizing this strategy, the color slice images of the Visible Human body can be continuously and serially segmented. This method is not complex but is rapid and automatic with less manual participation. The experimental results show that the primary organs of the Visible Human body can be accurately extracted.

The topography and morphometrics of the pubic ligaments

BackgroundConflicting anatomical reports and the little attention given to the pubic ligaments impede the interpretation of radiological and clinical examinations on groin pain. Morphometric data on the pubic ligaments are lacking. MethodsThe muscular relations of the symphysis pubis were examined in layered dissection (n = 10), hemipelves (n = 60) and (un)stained plastinated body slices of body donors (n = 3). The sagittal and coronal areas, width, mean and maximum thickness of pubic ligaments were determined. ResultsThe adductor longus, brevis, rectus abdominis and pyramidalis muscles are attached to the anterior pubic ligament (APL). The adductor brevis and gracilis muscle are connected to the inferior pubic ligament (IPL). The IPL and superior pubic ligament (SPL) are thicker than the APL and posterior pubic ligament (PPL). The PPL is the thinnest pubic ligament. The APL has a larger sagittal area in women than in men compared to the IPL. The SPL and IPL are thicker in men compared to women. ConclusionThe APL is the ligamentous anchor for the originating and inserting muscles. Investigations of the pubic ligaments might help to determine symphysis instability or severity of injury and should be included as a further criterion for surgical management.

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 3mm. Impregnation mixture is made of epoxy E12 resin plus E1 hardener (100ppw; 28ppw). 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 20-24hr. 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.

Ligamental compartments and their relation to the passing spinal nerves are detectable with MRI inside the lumbar neural foramina.

Intraforaminal ligaments (IFL) in lumbar neural foramina (NF) and their relation to the lumbar spinal nerves (SN) are addressed. Giemsa- and PAS-stained plastinated body slices of 15 lumbar spines were made and compared to MRI and CT data acquired of the same fresh specimens. We dissected one fixed lumbar spine to discuss our results with previous literature. Macroscopic pathophysiological changes and operational interventions at these lumbar spines were excluded. In the NF, thin medial IFL touch the SN. As a second compartment, intermedial vertical IFL are seen. A third lateral horizontal compartment of IFL is formed by thick cranial and caudal ligaments. Ligaments of the second and third compartments have no direct contact with the SN. From medial to lateral, the IFL thicken. All compartments are 3D reconstructed. If compartments of the IFL have no direct contact with the SN seen in the slices, a connection was noticed after dissection. Manual dissection seems to be inappropriate for a detailed study of the IFL. The lateral and intermedial compartments being free of the SN may transmit power and protect the SN, while the thin medial IFL may lead the SN passing the NF under physiological conditions. We conclude from the close topographical relation that the IFL may be relevant in foraminal stenosis. Any herniation in the NF presses IFL to the SN. Therefore, we think the IFL themselves could cause neurogenic claudication in case of their non-physiological turnover. Visualisation of IFL seems to be possible by using MRI. These slides can be retrieved under Electronic Supplementary Material.

Ligamental compartments and their relation to the passing spinal nerves are detectable with MRI inside the lumbar neural foramina

Intraforaminal ligaments (IFL) in lumbar neural foramina (NF) and their relation to the lumbar spinal nerves (SN) are addressed. Giemsa- and PAS-stained plastinated body slices of 15 lumbar spines were made and compared to MRI and CT data acquired of the same fresh specimens. We dissected one fixed lumbar spine to discuss our results with previous literature. Macroscopic pathophysiological changes and operational interventions at these lumbar spines were excluded. In the NF, thin medial IFL touch the SN. As a second compartment, intermedial vertical IFL are seen. A third lateral horizontal compartment of IFL is formed by thick cranial and caudal ligaments. Ligaments of the second and third compartments have no direct contact with the SN. From medial to lateral, the IFL thicken. All compartments are 3D reconstructed. If compartments of the IFL have no direct contact with the SN seen in the slices, a connection was noticed after dissection. Manual dissection seems to be inappropriate for a detailed study of the IFL. The lateral and intermedial compartments being free of the SN may transmit power and protect the SN, while the thin medial IFL may lead the SN passing the NF under physiological conditions. We conclude from the close topographical relation that the IFL may be relevant in foraminal stenosis. Any herniation in the NF presses IFL to the SN. Therefore, we think the IFL themselves could cause neurogenic claudication in case of their non-physiological turnover. Visualisation of IFL seems to be possible by using MRI. These slides can be retrieved under Electronic Supplementary Material.

Tracking morphological complexities of organ development in culture

Organogenesis is one of the most striking process during development. During this period, organ primordia pass throughout several stages in which the level of organisation increases in complexity to achieve the final organ architecture. Organ culture, a method in which an isolated organ is explanted and maintained ex-vivo, is an excellent tool for following the morphological dynamics during development. While most of the work has been made in early stages of development, culturing organs in mid-late stages is needed to understand the achievement of the final organ anatomy in the new-born. Here, we investigated the possibility of following morphological changes of the mice heart, lung, kidney and intestine using a filter-grid culture method for 7 days starting at E14.5. We observed that the anatomy, histology and survival of the cultured organs were indicative of a continuity of the developmental processes: they survived and morphodifferentiated during 5–7 days in culture. The exception was the heart, which started to die after 4 days. Using a second approach, we demonstrated that heart tissue can be easily cultured in body slices, together with other tissues such as the lung, with a healthier differentiation and longer survival. The culture method used here, permits a high-resolution imaging to identify the dynamic of organ architecture ex-vivo using morphovideos. We also confirmed the suitability of this system to perform lineage tracing using a vital dye in branching organs. In summary, this work tested the feasibility of monitoring and recording the anatomical changes that establish the final organ structure of the heart, lung, kidney and intestine. Additionally, this strategy allows the morphological study of organ development including fate maps with a relative long-term survival up to the onset of differentiation. This work contributes to elucidating how organs are formed, promoting the understanding of congenital malformations and to design organ replacement therapies.

Periodic acid-Schiff (PAS) reaction and plastination in whole body slices. A novel technique to identify fascial tissue structures

Since collagen rich fascial tissue is often very delicate and difficult to discern on native tissue slices, we have developed a method for staining full-body slices using the periodic acid-Schiff (PAS) reaction with subsequent plastination. Since the PAS reaction primarily stains carbohydrates, we could exploit the circumstance that different collagen types vary in carbohydrate content. Contrary to fasciae, tissues such as muscle, bone, nerves and blood vessels exhibit significantly less staining or remain unstained. We have validated the whole-body slice staining results in microscopic tissue slides which were stained with standard extracellular matrix stains such as Masson-Goldner trichrome stain and van-Gieson stain. Furthermore, we have performed immunofluorescence imaging to confirm the presence of collagen in the stained tissue. We achieved very good staining and plastination results and were able to clearly identify even very thin fascia in transversal body slices. This technique may prove useful in advancing our knowledge on the complex topography of fascial structures.

Hoffen P45: A Modified Polyester Plastination Technique for Both Brain and Body Slices

Hoffen P45: A Modified Polyester Plastination Technique for Both Brain and Body Slices

The Human Carotid Body Gene Expression and Function in Signaling of Hypoxia and Inflammation.

Although animal carotid body oxygen sensing and signaling has been extensively investigated, the human carotid body remains essentially uncharacterized. Therefore, we aimed to study the human carotid body in terms of morphology, global and specific expression of sensing and signaling genes as well as inflammatory genes. The human carotid body response to brief or prolonged hypoxia was studied in carotid body slices from adult surgical patients and ACh, ATP and cytokine release was analyzed. We demonstrate that the human carotid body expresses key oxygen sensing and signaling genes in similarity with animal carotid bodies with a few diverging data. The human carotid body moreover shows enrichment of genes in the inflammatory response and releases pro and anti-inflammatory cytokines in response to prolonged hypoxia. In response to acute hypoxia the human carotid body releases ACh and ATP and we thus translate previous findings in animal models to human tissue. We conclude that by releasing pro- and anti-inflammatory cytokines during hypoxia the human carotid body displays a structural and functional capacity to participate in sensing and mediating systemic inflammation.

Plastination: basic principles and methodology

Plastination is the method of long term preservation of the biological tissues with completely visible surface and high durability.The plastinates are devoid of harmful effects of formalin and they serve as excellent teaching tools in education. Additionally,plastination is an outstanding tool to study cross-sectional anatomy. The three major methods used in plastination are siliconeplastination, sheet plastination with epoxy method and sheet plastination with polyester method. Silicone plastination is the mostversatile technique which can be used for the cadavers, organs, portions and slices. Fresh or formalin-fixed (embalmed) specimens can be plastinated with this technique. More flexible specimens can be obtained if fresh tissues are preferred. Silicone plastination consists of five main steps. These are the preparation of the specimen (dissection and fixation if necessary), dehydration,defatting (degreasing), forced impregnation and curing (hardening). Epoxy plastination preserves 2-5 mm slices of biological tissues by using epoxy resins. In this technique, all tissue fluid and a significant amount of fatty tissue is replaced with a curableepoxy resin mixture. Epoxy plastination method provides precise semi-transparent sectional specimens and in these preparations;gross anatomical structures can be examined with the naked eye in a superb quality down to a sub macroscopic level. Polyesterplastination and classic silicone plastination techniques utilize the similar basic principles. In polyester plastination method, thetissue fluid is removed and is replaced with a curable polyester resin. This method can be used for head brain and body slices

The human carotid body releases acetylcholine, ATP and cytokines during hypoxia.

Studies on experimental animals established that the carotid bodies are sensory organs for detecting arterial blood O2 levels and that the ensuing chemosensory reflex is a major regulator of cardiorespiratory functions during hypoxia. However, little information is available on the human carotid body responses to hypoxia. The present study was performed on human carotid bodies obtained from surgical patients undergoing elective head and neck cancer surgery. Our results show that exposing carotid body slices to hypoxia for a period as brief as 5 min markedly facilitates the release of ACh and ATP. Furthermore, prolonged hypoxia for 1 h induces an increased release of interleukin (IL)-1β, IL-4, IL-6, IL-8 and IL-10. Immunohistochemical analysis revealed that type 1 cells of the human carotid body express an array of cytokine receptors as well as hypoxia-inducible factor-1α and hypoxia-inducible factor-2α. Taken together, these results demonstrate that ACh and ATP are released from the human carotid body in response to hypoxia, suggesting that these neurotransmitters, as in several experimental animal models, play a role in hypoxic signalling also in the human carotid body. The finding that the human carotid body releases cytokines in response to hypoxia adds to the growing body of information suggesting that the carotid body may play a role in detecting inflammation, providing a link between the immune system and the nervous system.

Plastination and its importance in teaching anatomy. Critical points for long-term preservation of human tissue.

Most medical curricula rely on human bodies for teaching macroscopic anatomy. Over the past 20 years, plastination has become an important means of preservation of organs, for well dissected specimens or for body slices. Here, several critical points regarding body donation with legal and ethical considerations for long-term preservation, the use of cadavers in teaching and the preparation of plastinates as an additional teaching tool will be discussed. Silicone S10 is the gold standard in the preparation of plastinates. An important point to respect is the preparation of specimens, since only very well dissected body parts or excellent tissue sections should be plastinated to show the extraordinary aspects of the human anatomy. The preparation of thin and transparent sections and preservation with P40 polyester provides an additional technique to prepare resistant body slices. A selection of samples prepared by S10 and P40 are shown and compared. In addition, Prussian or Berlin blue staining of brain slices is shown to discriminate better between gray and white matter and demonstrate neuroanatomical structures. These plastinates have been used for many years in teaching first- and second-year medical students and have not lost their appeal. Students and staff appreciate the use of such plastinates. One of the advantages is that their use is not restricted to the dissection hall; slices and body parts can be used in any lecture room or in small group teaching. Therefore, ethical and legal questions need to be addressed regarding their specific use. Plastinates do not replace the traditional dissection courses, since students learn best the anatomical features of a given region by hands-on dissection and by exploratory anatomy. Furthermore, plastinates are more rigid and do not allow demonstration of hidden structures; they also become more cumbersome for endoscopy or are too rigid for demonstrating mechanical features of joints. However, although not a replacement for traditional dissections, plastination provides an additional tool for long-term preservation and for teaching human anatomy.

Functional Fluorescently Labeled Bithiazole ΔF508-CFTR Corrector Imaged in Whole Body Slices in Mice

We previously reported the identification and structure-activity analysis of bithiazole-based correctors of defective cellular processing of the cystic fibrosis-causing CFTR mutant, ΔF508-CFTR. Here, we report the synthesis and uptake of a functional, fluorescently labeled bithiazole corrector. Following synthesis and functional analysis of four bithiazole-fluorophore conjugates, we found that 5, a bithazole-based BODIPY conjugate, had low micromolar potency for correction of defective ΔF508-CFTR cellular misprocessing, with comparable efficacy to benchmark corrector corr-4a. Intravenous administration of 5 to mice established its stability in extrahepatic tissues for tens of minutes. By fluorescence imaging of whole-body frozen slices, fluorescent corrector 5 was visualized strongly in gastrointestinal organs, with less in lung and liver. Our results provide proof-of-concept for mapping the biodistribution of a ΔF508-CFTR corrector by fluorophore labeling and fluorescence imaging of whole-body slices.

Quantification of human body fat tissue percentage by MRI

The MRI-based evaluation of the quantity and regional distribution of adipose tissue is one objective measure in the investigation of obesity. The aim of this article was to report a comprehensive and automatic analytical method for the determination of the volumes of subcutaneous fat tissue (SFT) and visceral fat tissue (VFT) in either the whole human body or selected slices or regions of interest. Using an MRI protocol in an examination position that was convenient for volunteers and patients with severe diseases, 22 healthy subjects were examined. The software platform was able to merge MRI scans of several body regions acquired in separate acquisitions. Through a cascade of image processing steps, SFT and VFT volumes were calculated. Whole-body SFT and VFT distributions, as well as fat distributions of defined body slices, were analysed in detail. Complete three-dimensional datasets were analysed in a reproducible manner with as few operator-dependent interventions as possible. In order to determine the SFT volume, the ARTIS (Adapted Rendering for Tissue Intensity Segmentation) algorithm was introduced. The advantage of the ARTIS algorithm was the delineation of SFT volumes in regions in which standard region grow techniques fail. Using the ARTIS algorithm, an automatic SFT volume detection was feasible. MRI data analysis was able to determine SFT and VFT volume percentages using new analytical strategies. With the techniques described, it was possible to detect changes in SFT and VFT percentages of the whole body and selected regions. The techniques presented in this study are likely to be of use in obesity-related investigations, as well as in the examination of longitudinal changes in weight during various medical conditions.

Anatomic and Dosimetric Changes During the Treatment Course of Intensity-Modulated Radiotherapy for Locally Advanced Nasopharyngeal Carcinoma

Many patients with nasopharyngeal carcinoma (NPC) have marked anatomic change during intensity-modulated radiation therapy (IMRT). In this study, the magnitude of anatomic changes and its dosimetric effects were quantified. Fifteen patients with locally advanced NPC treated with IMRT had repeated computed tomography (CT) after 18 fractions. A hybrid plan was made to the anatomy of the second computed tomography scan. The dose of the original plan, hybrid plan, and new plan were compared. The mean volume of left and right parotid decreased 6.19 mL and 6.44 mL, respectively. The transverse diameters of the upper bound of odontoid process, the center of odontoid process, and the center of C2 vertebral body slices contracted with the mean contraction of 8.2 mm, 9.4 mm, and 7.6 mm. Comparing the hybrid plan with the treatment plan, the coverage of target was maintained while the maximum dose to the brain stem and spinal cord increased by 0.08 to 6.51 Gy and 0.05 to 7.8 Gy. The mean dose to left and right parotid increased by 2.97 Gy and 2.57 Gy, respectively. A new plan reduced the dose of spinal cord, brain stem, and parotids. Measurable anatomic changes occurring during the IMRT for locally advanced NPC maintained the coverage of targets but increased the dose to critical organs. Those patients might benefit from replanning.

Low glucose effects on rat carotid body chemoreceptor cells' secretory responses and action potential frequency in the carotid sinus nerve

Glucose deprivation (hypoglycaemia) is counterbalanced by a neuroendocrine response in order to induce fast delivery of glucose to blood. Some central neurons can sense glucose, but nevertheless the most important glucose sensors/glycaemia regulators are located outside the brain. Some recent experimental evidence obtained in carotid body (CB) slices and isolated chemoreceptor cells in culture supports a role for the CB in glucose sensing and presumably glucose homeostasis, but this role has been questioned on the basis of a lack of effect of low glucose on the carotid sinus nerve activity. This work was performed in an attempt to clarify if low glucose is or is not a stimulus for the rat CB chemoreceptors. Using freshly isolated intact CB preparations we have monitored the release of catecholamines (CAs) and ATP from chemoreceptor cells in response to several concentrations of glucose, as indices of chemoreceptor cell sensitivity to glycaemia, and the electrical activity in the carotid sinus nerve (CSN), as an index of reflex-triggering output of the CB. We have observed that basal (20% O(2)) and hypoxia (7 and 10% O(2))-evoked release of CAs was identical in the presence of normal (5.55 mm) and low (3, 1 and 0 mm) glucose concentrations. 0 mm glucose did not activate the release of ATP from the CB, while hypoxia (5% O(2)) did. Basal and hypoxia (5% O(2))-induced CSN action potential frequency was identical with 5.55 and 1 mm glucose. Our results indicate that low glucose is not a direct stimulus for the rat carotid body chemoreceptors.

Using Plastinated Specimens in Teaching Veterinary Anatomy

Teaching veterinary anatomy continually presents new challenges. Cadaver procurement and usage comes under question from students, administration and the community. Student expectations rise while there participation wanes. Grades are hopelessly inflated. Time has been sliced away from many basic science courses. What can we do? We can run away or devise a plan. We use a tool that can make all these problems seem worse, it will take more of your time and your Dean's money, it will make some students angry with you, but it is a great hobby for those who enjoy preparing anatomical specimens. Plastination! Our lab has been using plastination for 20 years. Plastination was invented by Dr. Günter von Hagens 26 years ago at the University of Heidelberg. The silicone plastination process has four major steps. For a laboratory to be successful and remain even after you retire or get fired, the first step in the process must be done meticulously. Step 1: Specimen preparation and fixation ‐ plan what the specimen should demonstrate. Hollow organs must be dilated and fixed in a dilated form. Specimens must be positioned in an anatomical arrangement. Step 2: Dehydration: Cold (‐25°C) Acetone is preferred, however alcohol may be used. Step 3: Impregnation: Replacing the volatile solvent (acetone) with a curable polymer. Step 4: Curing, hardening, or cross‐linking the polymer ‐ also a critical step. You must know what the final product is to look like. Once cured, the specimen will remain in that position. Silicone plastination yields somewhat life‐like, dry, durable, aesthetically pleasing specimens that can be used in most any circumstance and any audience. Labelling specimens and/or photographing the specimens and labelling the photograph will enhance student use. Usage: A. Anatomical instruction ‐ Two formats have been used. 1. Plastinated specimens used with no wet dissection of that region and 2. Specimens used as a supplement and review after dissection has been completed. B. Client or public education. C. Anatomical review for Veterinarians. D. Display of anatomy area for tours. Benefits: A. Medical students and Practitioners ‐ Specimens are dry, therefore they are convenient to use at any time even on short notice. Items that are difficult to dissect and/or visualize are good adjuncts to teaching. B. Client/Public education ‐ Great public relations as well as seeing the involved anatomy. C. Anatomy program ‐ The use of prosected, plastinated large animal specimens allows students access to competently prepared, long lasting study material. By not having to dissect the entire large animal cadaver, students do not rush through dissection of the equine and bovine species limbs to compensate for the reduction in laboratory time. This approach allows sufficient time for students to study the limbs of various species within the time frame dictated by the curricular revision. Disadvantage: Some students may not do a complete dissection and/or not use their wet specimens for study/review. Some students want everything to be plastinated so they can memorize one example and forget about variation. Polyester and epoxy polymers may also be used for plastination. These polymers yield are used for preparing thin body slices, which are not flexible. With the advent of MRI, CT and ultrasonography an understanding of sectional anatomy is necessary, these thin slices are an ideal tool to aid understanding images of these modalities. Plastination is a fabulous tool, which is recommended as an adjunct for teaching anatomy.

Sex differences in whole body skeletal muscle mass measured by magnetic resonance imaging and its distribution in young Japanese adults

Objectives: To determine sex differences in the distribution of regional and total skeletal muscle (SM) using contiguous whole body magnetic resonance imaging (MRI) data, and to examine the relations between...

Studies on glomus cell sensitivity to hypoxia in carotid body slices.

Oxygen sensing is a fundamental biological process present in almost all life forms (for reviews see Bunn and Poyton, 1996; Lopez-Barneo et al., 2001). In mammals, the survival in acute hypoxia requires fast respiratory and cardiocirculatory adjustments to ensure sufficient O2 supply to the most critical organs such as the brain or the heart. The main O2 sensor mediating the acute responses to hypoxia is the carotid body (CB), a minute bilateral organ at the bifurcation of the carotid artery innervated by afferent chemosensory fibers. In conditions of hypoxemia, the CBs stimulate the brainstem respiratory centers to evoke hyperventilation. Glomus, or type I, cells are electrically excitable (Duchen et al., 1988, Lopez-Barneo et al., 1988) and constitute the major O2sensitive elements of the CB (Lopez-Barneo et al., 1988, Delpiano and Hescheler, 1989; Peers 1990; Stea and Nurse, 1991, Buckler, 1997). It is broadly accepted that hypoxia signaling in these cells requires the inhibition of O2- sensitive potassium channels of the plasma membrane, which leads to depolarization, external Ca2+influx, and release of the transmitters that activate the afferent sensory fibers (Lopez- Barneo et al., 1993; Urena et al., 1994; Buckler and Vaughan-Jones, 1994; Carpenter et al., 2000; Pardal et al., 2000). This basic scheme of chemotransduction has also been proposed to operate in other 02-sensitive neurosecretory systems, such as cells in the lung neuroepithelial bodies (Youngson et al., 1993), chromaffin cells of the adrenal medulla (Thompson and Nurse, 1998), or PC-12 cells (Zhu et al., 1996).