Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Plastic Embedding”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 991 records · Page 55Linked to original sources
PubMed1998

The effect of hyperbaric oxygen treatment on bone tissue reactions to c.p. titanium implants placed in free autogenous bone grafts. A histomorphometric study in the rabbit mandible.

This study aimed to investigate the effect of hyperbaric oxygen (HBO) therapy on the tissue reactions to commercially pure (c.p.) titanium implants placed in free autogenous bone graft by a 1-stage procedure. Eighty c.p. titanium implants were placed in the bone grafted from iliac crest to bilateral mandible of 40 Japanese white rabbits without tapping. Twenty rabbits underwent daily a HBO treatment for 60 min under 2.4 ATA during 20 consecutive days and the other untreated 20 rabbits served as controls. The implants with surrounding bone tissue were retrieved 20, 30, 60, 90 and 120 days after surgery, fixated, dehydrated and embedded in resin. About 20 microns thick ground sections were prepared prior to microscopical observations. The bone area and the bone-to-implant contact inside the threads were calculated separately in the grafted bone and in the host bone for each implant. After 30 and up to 120 days, the HBO treated group showed more bone-to-implant contacts in the grafted bone as compared to the non-HBO treated group. In the host bone there were no differences observed between HBO and non-HBO treated groups. This study indicated that HBO treatment was beneficial for the tissue incorporation of c.p. titanium implants when placed immediately in free autogenous bone grafts.

Animals↗
PubMed1995

An easy method for the removal of Epon resin from semi-thin sections. Application of the avidin-biotin technique.

A simple procedure is described for removing Epon resin from semi-thin 1 micron sections, which permits excellent postembedding immunohistochemical staining (avidin-biotin complex technique). The procedure was developed for the detection of growth hormone and prolactin in bovine adenohypophysis fixed with 2% paraformaldehyde and 0.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.4-7.6. The results indicate that the removal of the epoxy embedding medium prior to the application of the immunohistochemical reagents was essential for the successful localization of the antigenic determinants of the two hormones. The immunocytochemical reactivity was obtained only after treating the sections with a solution of potassium hydroxide in a mixture of absolute methyl alcohol and propylene oxide (Maxwell's solution). An enhanced immunoreactivity was obtained when this treatment was followed by an additional treatment with either 4% hydrogen peroxide or a saturated aqueous solution of sodium metaperiodate. Because of the easy preparation of the Epon removal solution and the good structural preservation without damage to the antigenic determinants, Maxwell's solution is suggested as a good etching agent which can be used in immunohistochemical studies on semi-thin sections with excellent results.

Animals↗
PubMed1993

Three-dimensional cytoskeletal structures of the chinchilla organ of Corti: scanning electron microscopy application of the polyethylene glycol method.

We describe the application of a polyethylene glycol (PEG) embedding technique to examine the sensory and supporting structures of the inner ear. The chinchilla organ of Corti was exposed by cracking PEG embedded cochleas. A range of PEG molecular weights (2000-8000) were utilized; PEG 2000, with a melting point of 57 degrees C was preferred. After removal of the PEG, the three-dimensional aspects of intracellular structures were observed using scanning electron microscopy. Filamentous elements in the hair cell cuticular plate and in the supporting cells were clearly observed, as was the meshwork of cross-linked actin filaments in the cuticular portion of sensory hair cells. Microtubule and microfilament alignment patterns in pillar and Deiters cells were also clearly demonstrated. Characteristic structures in the outer hair cell synaptic region, such as the post-synaptic cistern and synaptic body, were well preserved using the PEG method.

Actin Cytoskeleton↗
PubMed1997

The mucosal barrier following abutment dis/reconnection. An experimental study in dogs.

In the present experiment, the effect on the marginal peri-implant tissues following repeated abutment removal and subsequent reconnection was studied. 5 beagle dogs were used. The mandibular premolars were extracted and 2 fixtures of the Brånemark System were installed, 1 in each mandibular quadrant. 3 months later, abutment connection was performed. A 6-month period of plaque control was initiated. Once a month during the plaque control period, the abutment of the right side (test) in each dog was disconnected, cleaned and reconnected to the failure. Thus, each test abutment was removed and reconnected altogether 5x during this period. The contralateral abutment remained undisturbed for 6 months and served as control. 1 month after the last reconnection, the animals were sacrificed and tissue samples, comprising the implant and the surrounding soft and hard peri-implant tissues, were obtained, decalcified, embedded in Epon and sectioned. The following landmarks were identified and used for linear measurements: PM (the marginal portion of the peri-implant mucosa), aJE (the level of the apical termination of the junctional epithelium), B (the marginal level of bone to implant contact), A/F (the abutment/fixture border). The findings indicate that the dis- and subsequent reconnections of the abutment component of the implant compromised the mucosal barrier and resulted in a more "apically" positioned zone of connective tissue. The additional marginal bone resorption observed at the test sites following abutment manipulation may be the result of tissue reactions initiated to establish a proper "biological width" of the mucosal-implant barrier.

Alveolar Bone Loss↗
PubMed2005

Embedding of large specimens in glycol methacrylate: prerequisites for multi-signal detection and high-resolution imaging.

Acrylic resin mixtures are commonly used to study microscopic sections of biological specimens, giving the advantage of good morphological preservation. Existing embedding protocols, however, are suitable for tissue blocks, not exceeding 1 mm in thickness. We have developed a protocol to embed larger specimens (up to 2 cm(3)) in Technovit 8100. This medium allowed us to perform classic histological (trichrome), silver, as well as immunohistochemical staining, needed for multi-signal detection at high-resolution imaging to reconstruct a three-dimensional interpretation of a serially sectioned muscle. The technique was applied to reconstruct the semitendinosus muscle of a fetal pig, 44 days post conception, featuring connective tissue, intramuscular nerves, blood vessels, and muscle fibre types. For the reconstruction, a technique was used that enabled us to insert high-resolution images of histological details into low-resolution images of the entire muscle.

Animals↗
PubMed1993

A new subdivision of the rat midbrain periaqueductal gray based on its myeloarchitecture.

A new subdivision of the periaqueductal gray (PAG) based on its myelin content is described. Using a methacrylate-resin embedding technique for high resolution light microscopy, a myeloarchitectonic map of the normal rat PAG was made. Six main columns were distinguished. Column I is the central thin subependymal layer, in which myelinated axons are virtually absent. This layer is surrounded by column II with many rostrocaudally oriented fibers. Lateral and dorsolateral to column II lies column III, which contains rostrocaudally as well as tangentially oriented fibers. Column III is divided into a ventral and a dorsal part by column IV. This column is located in the dorsolateral part of the PAG and contains very few, mostly tangentially oriented fibers. Column V is located ventrally and contains a mixture of rostrocaudally and tangentially oriented fibers. Column VI is only present in the most caudal part of the PAG and contains mainly tangential fibers.

Animals↗
PubMed1991

Histochemical localisation of acid phosphatase and non-specific esterase in the midguts of two species of tick, Boophilus microplus and Rhipicephalus appendiculatus, as determined by light microscopy.

Serial sections of glycol methacrylate-embedded and frozen midguts of Boophilus microplus and Rhipicephalus appendiculatus were studied histochemically by light microscopy. The use of the naphthol AS-TR phosphate technique combined with glycol methacrylate embedding enabled the precise localisation of lysosomal enzyme activity, despite the ubiquity of haematin granules in tick midgut epithelia. The presence of acid phosphatase and non-specific esterase activity in the same cells was observed in all of the various developmental stages and feeding phases of the ticks. The pattern of appearance of these cells paralleled the reported level of protease activity in the midgut lumen. The cells were found to be solitary, particularly during the slow digestive phases, and appeared to move into the lumen, where they eventually disintegrated. The cells therefore appear to function as holocrine secretory cells. This is the first report indicating the presence of such secretory cells in the midgut of unfed ticks. The disintegration of these cells in the lumen suggests that lumenal digestion may be more important than hitherto realised.

Acid Phosphatase↗
PubMed2001

Enhancement of immunogold-labelled focal adhesion sites in fibroblasts cultured on metal substrates: problems and solutions.

Visualisation of cell adhesion patterns by scanning electron microscopy requires special preparation and labelling. The membranes and cytoplasm must be removed, without damaging the antigen, to facilitate antibody access to vinculin in the focal adhesions. Low beam energy imaging is used to visualise the cell undersurface (embedded in resin after staining with osmium tetroxide) and immunogold-labelled adhesion sites. The gold probe, must be large enough (>40 nm) for detection, while viewing the whole cell, but large gold markers increase steric hindrance and decrease labelling efficiency. This problem can be overcome by using small gold probes (1-5 nm) followed by enlargement with silver enhancement, but osmium tetroxide stain etches the silver. We demonstrated that metal substrates increased this etching. Reducing the concentration of osmium tetroxide and incubation time reduced the amount of etching. We have demonstrated that gold enhancement was not etched by osmium tetroxide, irrespective of the substrate. Therefore, comparative studies of cell adhesion to different biomaterial substrates can be performed using immunogold-labelling with gold enhancement.

Acrylic Resins↗
PubMed2000

Use of a Technovit 7200 VLC to facilitate integrated determination of aluminum by light and electron microscopy.

Technovit 7200 VLC is an excellent embedding medium for both inorganic histochemistry by light microscopy and X-ray microanalysis by scanning and transmission electron microscopy. Liver samples from rats after intraperitoneal treatment with aluminum chloride were fixed in glutaraldehyde and embedded in the resin. Thick sections were easily cut on an ultramicrotome and stained with aluminon for aluminum (Al). An intense positive reaction with aluminon was observed in the Kupffer cells by light microscopy. The surface structures of the same resin block cut for light microscopy were observed under a scanning electron microscope fitted with an energy dispersive X-ray spectrometer. The Kupffer cells appeared white in the backscattered mode. Localization of Al in the Kupffer cells was confirmed by an X-ray distribution map in the scanning electron microscope. Subcellular localization of Al in the Kupffer cells was performed on the same semithin sections using a transmission electron microscope equipped with an energy dispersive X-ray spectrometer. Most Al was found in lysosomes of the Kupffer cells. The resin was stable in the electron beam and chlorine-free.

Acrylic Resins↗
PubMed1994

The organization of subperitoneal connective tissue in the female pelvis.

The organization of the subperitoneal connective tissue in the female pelvis was studied in 6 cases, aged between 58 and 65, deceased from extrapelvic diseases. The pelvic viscera were removed as a whole with the surrounding subperitoneal tissue using a circular cut performed at the level of the pelvic walls. The specimens were fixed in 5% formalin. The bladder dome, uterine body and rectum, up to the anorectal junction, were removed. In two cases the specimens were dehydrated and embedded in paraffin. Ten micron sections were stained with Haematoxilin-Eosin and Azan-Mallory. In 2 other cases the specimens were plastinated according to Von Hagens E12 technique; the passage in acetone at room temperature lasted 24 hours. In the remaining 2 cases the plastination with E12 was carried out after a passage in acetone at room temperature which lasted about 15 days in order to fully remove the lipids from the subperitoneal adipose tissue. The morphological findings demonstrate that the subperitoneal tissue of the female pelvis has an areolar structure with small adipose lobules separated by thin fibrous laminae connected to the vasculo-nervous bundles and to the parietal and visceral layers of the fascia pelvica. The three dimensional spatial organization of the network of thin connective laminae constitutes an anatomical device with possible supporting properties.

Connective Tissue↗
PubMed1999

Analysis of cell death in the trochlear nucleus of the chick embryo: calibration of the optical disector counting method reveals systematic bias.

Detection of changes in numbers of neurons is essential for an understanding of neuronal development, function, and death. Optical disector counting is claimed to be the most efficient technique to estimate accurate numbers of neurons in microscopic sections. We calibrated the optical disector by comparison with three-dimensional reconstructions from serial sections and determined how accurate this technique is relative to conventional profile counting methods. The calibration was performed on the trochlear nucleus in developing chicks. Optical disector estimates, when obtained as generally recommended, were about 25% lower than the actual number of neurons. This underestimate was caused by a nonuniform (bimodal) distribution of neuronal nuclei in paraffin and plastic (glycolmethacrylate) sections, but not in cryosections. The density of neurons in the core of the paraffin and plastic sections was substantially lower than in the upper and lower margins of these sections. Accurate estimates of neuronal numbers were obtained with a modified optical disector method that sampled the entire extent of tissue sections. Previous estimates of numbers of trochlear neurons in the developing chick have been controversial. The modified (calibrated) optical disector method revealed that the number of trochlear neurons decreased from about 1,600 at day 8.5 of incubation (embryonic day, [E] 8.5) to about 900 at the time of hatching. Numbers of pyknotic nuclei peaked at E6 and at E9, revealing an additional early, but postproliferative, period of cell death. Taken together, these data emphasize the need for calibration of stereological counting techniques and the need to examine sampling strategies for potential bias.

Animals↗
PubMedDate not supplied

Morphologic and radioautographic studies of bone formation in relation to titanium implants using the rat tibia as a model.

A rat tibia model was developed to analyze bone formation leading to osseointegration with threaded titanium implants. Miniaturized titanium implants were placed in the anterior aspect of the upper tibia of rats weighing 350 g. Twenty-four rats were involved; 12 rats were sacrificed at 6 weeks, and another two rats were sacrificed weekly for 6 weeks following implantation. Four days prior to sacrifice, the animals were injected intraperitoneally with 3H-proline (10 microCi/g body weight). The rats were fixed by perfusion with 5% glutaraldehyde, and the tibia were decalcified and embedded in Epon. The implants were removed from the embedded bone by fracturing, and the specimens re-embedded. Sections were prepared for light and electron microscopy and radioautography. Morphologic observations indicated that implant placement resulted in bone necrosis and resorption. This was followed by bone growth from the old bone surfaces filling the threads with vascular channels and new lamellar bone. Osseointegration was complete at 6 weeks in all animals examined. Electron microscopy at various places along the bone-titanium interface showed an amorphous layer, a granular electron-dense layer, or a layer of uncalcified collagen fibrils. At each week after surgery, radioautography showed the position and thickness of new bone labeled with 3H-proline during the last 4 days. Radiolabeled new bone was deposited only on previously existing bone and extended toward the available space. Since there was no apparent relationship between the implant and the new bone, it was suggested that titanium is biocompatible, but not necessarily osteoinductive.

Animals↗
PubMed1992

Reevaluation of envelope profiles and cytoplasmic ultrastructure of mycobacteria processed by conventional embedding and freeze-substitution protocols.

The cell envelope architectures and cytoplasmic structures of Mycobacterium aurum CIPT 1210005, M. fortuitum, M. phlei 425, and M. thermoresistible ATCC 19527 were compared by conventional embedding and freeze-substitution methods. To ascertain the integrity of cells during each stage of the processing regimens, [1-14C]acetate was incorporated into the mycolic acids of mycobacterial walls, and the extraction of labeled mycolic acids was monitored by liquid scintillation counting. Radiolabeled mycolic acids were extracted by both processing methods; however, freeze-substitution resulted in the extraction of markedly less radiolabel. During conventional processing of cells, most of the radiolabel was extracted during the dehydration stage, whereas postsubstitution washes in acetone yielded the greatest loss of radiolabel during freeze-substitution. Conventional embedding frequently produced cells with condensed fibrous nucleoids and occasional mesosomes. Their cell walls were relatively thick (approximately 25 nm) but lacked substance. Freeze-substituted cells appeared more robust, with well-dispersed nucleoids and ribosomes. The walls of all species were much thinner than those of their conventionally processed counterparts, but these stained well, which was an indication of more wall substance; the fabric of these walls, in particular the plasma membrane, appeared highly condensed and tightly apposed to the peptidoglycan. Some species possessed a thick, irregular outer layer that was readily visualized in the absence of exogenous stabilizing agents by freeze-substitution. Since freeze-substituted mycobacteria retained a greater percentage of mycolic acids in their walls, and probably other labile wall and cytoplasmic constituents, we believe that freeze-substitution provides a more accurate image of structural organization in mycobacteria than that achieved by conventional procedures.

Cell Membrane↗
PubMedDate not supplied

Postembedding ultrastructural in situ hybridization on ultrathin cryosections and LR white resin sections.

A method was developed for nonisotopic postembedding in situ hybridization (ISH) on ultrathin sections of frozen and of LR White resin-embedded material at the electron microscopic level. The method was successfully applied to detect Epstein-Barr virus (EBV) DNA in the P3HR1 human Burkitt's lymphoma cell line. Each of the steps in the procedure had to be optimized for successful ISH on the frozen and LR White sections. The most important conditions are described. Predigestion with proteinase K was only necessary with the resin sections. Sections were treated with sodium hydroxide to denature target DNA and were hybridized with a biotinylated probe. The probe was best detected with a primary antibody to biotin followed by a gold-conjugated secondary antibody. EBV DNA was detected in the nucleus and/or cytoplasm in 10% to 20% of P3HR1 cells. A similar percentage of cells in thin L-sectioned material prepared by routine methods showed virus particles at different stages of maturation.

Acrylic Resins↗
PubMed2000

Expanded polytetrafluoroethylene entubulation of the rabbit inferior alveolar nerve.

OBJECTIVE: The purpose of this study was to evaluate the use of an expanded polytetrafluoroethylene conduit in the treatment of a 6. 0-mm gap in the rabbit inferior alveolar nerve and compare the results with those of an autogenous interpositional tibial nerve graft. STUDY DESIGN: The inferior alveolar nerves of 5 adult New Zealand White female rabbits (10 nerves) were exposed bilaterally, and a 6-mm segment of each nerve was resected. On one side, chosen at random, the gap was immediately bridged through use of an 8.0 x 2. 0-mm expanded polytetrafluoroethylene conduit; on the other side, the gap was grafted with an autogenous tibial nerve graft. Two randomly selected nerves served as sham-dissected controls. At 15 weeks after surgery, the animals were killed and the entire nerve segments were harvested and prepared according to standard fixation and embedding techniques. The sections were examined histomorphometrically to quantify the degree of axonal regeneration through definition of fascicular number, total fascicular surface area, axonal density, and mean axonal diameter at 3 locations along the repair site. RESULTS: Light microscopic examination revealed the presence of disorganized neural tissue in both groups, with slightly more fibrovascular interfascicular tissue in the expanded polytetrafluoroethylene group. Histomorphometric analysis revealed no significant differences between groups for most of the measured variables. The mean axonal diameter varied between groups, and the fascicular number was greater in the expanded polytetrafluoroethylene group at the middle site. CONCLUSIONS: This study demonstrates that regeneration of the inferior alveolar nerve can occur across a 6.0-mm gap through an expanded polytetrafluoroethylene tube with results comparable to those of an autogenous nerve graft, significant donor site morbidity being avoided. The significant differences between groups were probably due to greater containment of regenerating axonal fibers in the expanded polytetrafluoroethylene group.

Animals↗
PubMed2002

Canal-centering ability of two rotary file systems.

The ability of two nickel-titanium rotary file systems to maintain the original path of the canal were compared by using a new split-mold design (the Endodontic Cube). The mesial roots of 10 mandibular first- and second-molar teeth with separate canals from orifice to foramen were divided into two groups based on canal curvature and morphology. Working lengths for each canal were determined 1.0 mm short of where the file tip was visible on the external surface of the root. The teeth were embedded in composite resin by using the Endodontic Cube as a mold and sectioned into five pieces. ProFile Ni-Ti.06 taper Series 29 and ProFile GT rotary instrumentation techniques were randomly assigned to one of the canals and used in a crown-down fashion to a 0.217 ProFile and a 0.06 GT master apical file, respectively. Digital photographs were made of the middle three sections and software features of Adobe Photoshop were used to measure the movement of the canal centers by superimposing the images of the instrumented and uninstrumented canals. Data were analyzed using ANOVA. Both systems remained centered within the canal with minimal deviation from the original canal path. The largest mean movement was less than 0.15 mm. The Endodontic Cube was an effective tool for studying and comparing instrumentation techniques.

Analysis of Variance↗
PubMed1993

Subpopulations of GABA neurons containing somatostatin, neuropeptide Y, and parvalbumin in the dorsomedial cortex of the lizard Psammodromus algirus.

Different subpopulations of GABA neurons containing the neuropeptides somatostatin and neuropeptide Y, and the calcium binding protein parvalbumin were studied by immunocytochemistry using light and electron microscopy in the dorsomedial cortex of the lizard Psammodromus algirus to investigate the connectivity of different subsets of GABA neurons in the lizard dorsomedial cortical circuitry and to compare cortical regions of reptiles and mammals. GABA neurons were classified into different subsets by using the peroxidase anti-peroxidase immunohistochemical method on adjacent Araldite-embedded semithin sections. GABA neurons in the dorsomedial cortex fall into three major subsets: 1) neurons with somatostatin (and neuropeptide Y), which accounted for about 44% of the GABA population; 2) neurons with parvalbumin, which accounted for about 13% of the GABA neurons; and 3) neurons without parvalbumin or neuropeptides, which represented 40% of all GABA cells. This division of GABA neurons in non-overlapping subpopulations of neuropeptide- and parvalbumin-containing cells is similar to that found in the mammalian hippocampal formation. On the basis of the nerve terminal fields, somatostatin- and parvalbumin-immunoreactive neuronal populations appear to be functionally different, acting on different portions of the projection neurons. Parvalbumin-immunoreactive neurons inhibit the pyramidal neurons at the cell body level, whereas somatostatin-immunoreactive neurons inhibit them on distal dendrites. The results of the present study add more similarities between the lizard dorsomedial cortex and parts of the mammalian hippocampus.

Animals↗
PubMed2000

Co-localization of multiple antigens and specific DNA. A novel method using methyl methacrylate-embedded semithin serial sections and catalyzed reporter deposition.

Co-localization of proteins and nucleic acid sequences by in situ hybridization and immunohistochemistry is frequently difficult as the process necessary to detect the target structure of one technique may negatively affect the target of the other. Morphological impairment may also limit the application of the two techniques on sensitive tissue. To overcome these problems we developed a method to perform in situ hybridization and immunohistochemistry on semithin sections of methyl methacrylate-embedded tissue. Microwave-stimulated antigen retrieval, signal amplification by catalyzed reporter deposition, and fluorescent dyes were used for both techniques, yielding high sensitivity and excellent morphological preservation compared to conventional paraffin sections. Co-localization of in situ hybridization and immunohistochemistry signals with high morphological resolution was achieved on single sections as well as on adjacent multiple serial sections, using computerized image processing. The latter allowed for the co-localization of multiple antigens and a specific DNA sequence at the same tissue level. The method was successfully applied to radiation bone marrow chimeric rats created by transplanting wild-type Lewis rat bone marrow into TK-tsa transgenic Lewis rats, in an attempt to trace and characterize TK-tsa transgenic cells. It also proved useful in the co-localization of multiple antigens in peripheral nerve biopsies.

Animals↗