Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Microtomy”

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 667 records · Page 37Linked to original sources
PubMed1991

Detection of p24 in HIV-1 infected cells embedded in LR White and Lowicryl K4M.

In this study we present a postembedding on-grid immunogold labelling procedure for the ultrastructural localization of the HIV-1 core protein p24. HIV-1 infected cells were fixed in 0.1% glutaraldehyde, incompletely dehydrated and embedded in LR White or in Lowicryl K4M. Antigenic sites were detected by incubation of ultrathin sections with primary mouse monoclonal antibody anti-HIV-1 p24, followed by the secondary antibody goat anti-mouse IgG coupled to 10nm gold particles. Antigenicity of p24 was found to withstand the applied fixation and was shown to be preserved in LR White as well as in Lowicryl. The described procedure permits the uncomplicated and easy detection of p24 in HIV-1 infected cells and tissues.

Acrylic Resins↗
PubMed1983

Distribution of different fibre types in human skeletal muscles. I. Method for the preparation and analysis of cross-sections of whole tibialis anterior.

The aim of this study was to examine whether small biopsy specimens are representative of the whole human skeletal muscle or whether the different fibre types are unevenly distributed at different depths of the muscle. Ten micrometre thick cross-sections of whole human tibialis anterior were prepared using LKB PMV Cryo-Microtomes with a stroke length of 160 to 480 mm and the sections were stained for myofibrillar ATPase according to a modified procedure. The total and relative number of different fibres (Types 1 and 2) was determined in every 9th mm2 of the section. The data obtained were analysed by means of a computer program, which allowed assessment of bivariate data in the form of contour plots. The total number of fibres varied greatly between individuals (from 96 000 to 162 000; five individuals). The relative number of different fibres varied systematically in all individuals as a function of depth in the muscle. There was a gradual, often dramatic, relative increase in Type 2 fibre occurrence from the surface of the muscle (about 10--25%) towards the deeper regions (30--50%), the maximum being approximately along a line slightly posterior to the middle of the muscle. Additionally, superficial peaks were seen in places. In conclusion, the fibre type distribution in the tibialis anterior is not random. These results point to the importance of defining biopsy depth.

Adolescent↗
PubMed1991

A histochemical method for the demonstration of unsaturated lipids.

A method has been developed for the histochemical demonstration of unsaturated lipids in light microscopy. It is a peracetic acid-thiocarbohydrazide-silver protein sequence followed by a physical development procedure. In the present study on paraffin and cryostat sections of liver, brain and ovary, unsaturated lipids were visualized as distinct reaction products coloured various shades of brown and black. The reaction products are easier to see and the method is more efficient than the peracetic acid-Schiff method.

Animals↗
PubMed1992

Enkephalin- and serotonin-like immunoreactivity in the aortico-pulmonary paraganglia of the white-belly opossum Didelphis albiventris (Marsupialia).

Serial histological sections of the interatrial septum and basal heart vessels of the weaned and juvenile white-belly opossum (Didelphis albiventris) were obtained in order to study the presence of paraganglia and their content of regulatory peptides and serotonin. Paraganglion groups were mapped between the aorta and pulmonary arteries and close to the bifurcation of the pulmonary trunk and were found to contain cells with immunoreactivity to serotonin and to the neuroendocrine markers PGP 9.5 and NSE. When these paraganglia were tested for immunoreactivity to a battery of regulatory peptides, all were found to be positive for methionine-enkephalin, leucine-enkephalin and galanin. The hypothesis is raised that these peptides and serotonin, besides catecholamines, produced by these paraganglia may play a physiological role in the functions of the cardiovascular system of the white-belly opossum.

Animals↗
PubMed1978

Fine structural analysis of the morphological changes involved in the blanching, cooking, dehydration and rehydration of green bean pod tissue.

The influence of blanching and cooking on green bean pod tissue (Phaseolus vulgaris var.nanus L.) was studied by transmission electron microscopy and characteristic histological and cytological effects were recorded. The analysis of dehydraded and rehydrated samples (dehydration by hot air, rehydration by boiling) showed that much of the shrinkage after dehydration can be compensated by water absorption during rehydration. The morphological appearance of boiled raw tissue, however, is not reached.

Cooking↗
PubMed1995

Localization of dopamine D2 receptor mRNA in glomus cells of the rabbit carotid body by in situ hybridization.

The localization of mRNA coding for the dopamine D2 receptor was studied in the rabbit carotid body using in situ hybridization with synthetic 35S-labelled oligodeoxynucleotides. Using autoradiography on cryostat or semi-thin sections, labelling was observed over the cytoplasm of glomus cells, but not over sustentacular cells. A quantitative study showed that labelling intensity (silver grain density) was increased by haloperidol treatment. These results suggest that glomus cells express the dopamine D2 receptor gene and that this expression is regulated.

Animals↗
PubMed1995

Fine-structural investigation of rat brain microvascular endothelial cells: tight junctions and vesicular structures in freshly isolated and cultured preparations.

A comparison was made between endothelial cells in freshly-isolated rat brain microvessels, and following culture of the cells for 1-10 days during growth to confluence. Attention focused on tight junctions and vesicular structures, as seen in thin sections and freeze-fracture replicas. Freshly-isolated vessels had an abnormal appearance, with a profusion of luminal microvillar processes, and extensive cytoplasmic vacuolation. There were numerous vesicular profiles, reaching a density of approximately 60 microns-2, and with a large proportion open to the surface, as shown by labelling with cationized ferritin at 4 degrees C for 5 min. Junctional zones were relatively loosely organized, with evidence for some cell:cell separation, as well as some residual tight junctional sites within zonula adhaerens junctions. In freeze-fracture replicas, junctional strands showed segments of tightly packed intramembrane particles, generally on the P face. After 1 day in culture, the cells appeared more normal, with no vacuolation or luminal processes. Vesicles were still numerous, some associated with junctional zones, while tight junctions were relatively sparse; freeze-fracture showed some incomplete tight junctional strands, with some of the intramembrane particles fracturing onto the E face. The double offset fibrillar nature of the strands could occasionally be seen. Cells cultured for 4 and 10 days showed a progressive increase in the completeness of the junctional zone, with more tight junctional contacts within the length of the adhaerens junction, and an aggregation of microfilaments in the underlying cytoplasm. The number of vesicular profiles declined, and they were progressively excluded from the junctional zone. These observations have relevance for studies on the physiology of the brain endothelium in vitro, and for comparisons with the in vivo condition.

Animals↗
PubMed1995

Immunocytochemical localization of opsin in rod photoreceptors during periods of rapid disc assembly.

Transport of opsin from photoreceptor inner to outer segments has been assumed to occur via the connecting cilium, the only permanent structural connection between these two regions. However, in prior work, little or no immunoreactive opsin has been detected in the cilium, despite the high rate of transport of this protein. This suggests that immune epitopes are masked during passage through the cilium or that opsin is transported via an extra-ciliary route. In this study, we stained the photoreceptors of Xenopus laevis with well-characterized monoclonal antibodies directed at the N-terminal, C-terminal, and 5-6 loop regions of bovine opsin. This was done on isolated retinas incubated in vitro under conditions that support rapid disc assembly, to insure that opsin transport to forming discs was occurring at the time of fixation. Five MAbs that gave robust staining of Xenopus rod inner segment/rod outer segment preparations with the light microscope were utilized for electron microscopic studies on LR White embedded or cryo-ultrathin sections. Four of these stained outer segment discs and inner segment vesicles and plasma membrane. However, no significant staining of the connecting cilium was found. Furthermore, freeze-fractured mouse photoreceptors prepared by the 'fracture-label' technique showed extensive labelling of membrane compartments but lacked staining of the connecting cilium. Isolated retinas incubated under conditions that support robust rod disc synthesis contained many finger-like and vesicular projections of the apical inner segment plasma membrane and inner segment vesicles extending into them. Rod outer segment nascent discs usually made close contact with the inner segment. Both the vesicular profiles associated with the inner segment plasma membrane and the basal discs extending to the inner segment were heavily stained with all four anti-opsin antibodies. This suggests an alternate route for bulk transport of opsin to newly forming discs that involves direct transfer from apical inner segment plasma membrane to nascent discs.

Animals↗
PubMed1991

Two types of sympathetic axon innervating the juxtaglomerular arterioles of the rabbit and rat kidney differ structurally from those supplying other arteries.

Ultrastructural analyses of serial thin sections have revealed two structurally different types of sympathetic axon innervating the afferent and efferent juxtaglomerular arterioles and the intralobular arteries in the outer cortex of the rabbit kidney. Both types of axon have also been found in association with an afferent arteriole in rat kidney. One axon type consists of relatively large diameter unmyelinated axons bearing varicosities in the form of slight expansions. The varicosities have a distinct structural zonation: synaptic vesicles occupy the expansion which faces the smooth muscle cells, whereas the rest of the axon is filled with numerous microtubules. The other axon type has varicosities containing vesicles and mitochondria but few microtubules. The varicosities are generally small and the intervaricosities very thin. The relationship of both axon types with support cells and/or basal lamina is sometimes poorly defined. Both axon types are catecholaminergic as their vesicles take up 6-hydroxydopamine and both types form junctions with arteriolar smooth muscle cells. As well as differing from each other, both types of intrarenal axon differ in several respects from those which innervate other arterial vessels.

Animals↗
PubMed1983

Synaptic vesicles and the nerve-muscle preparation in resinless sections.

Resinless sections have been used to study the fine structural organization of cellular organelles and fibrous components at synapses. Presynaptic elements of the neuromuscular junction, representing the P.N.S., and of cerebral cortical synapses, representing the C.N.S., are examined and compared. Typical ultrastructural features are apparent in presynaptic and postsynaptic components and in the surrounding muscle and neuronal tissue demonstrating the reliability and usefulness of the technique.

Animals↗
PubMed1995

Glutamate, gamma-aminobutyric acid and tachykinin-immunoreactive synapses in the cat nucleus tractus solitarii.

Neurophysiological and pharmacological evidence suggests that glutamate, gamma-aminobutyric acid and tachykinins (substance P and neurokinin A) each have a role in cardiovascular regulation in the nucleus tractus solitarii. This study describes the ultrastructural relationships between nerve terminals immunoreactive for these substances in the nucleus tractus solitarii of the cat using post-embedding immunogold (single and double) labelling techniques on sections of tissue embedded in LR White resin. The technique combines a high specificity of labelling with good ultrastructural and antigenic preservation. Glutamate-immunoreactive terminals, recognized by their high density of gold particle labelling compared to the mean tissue level of labelling, accounted for about 40% of all synaptic terminals in the region of the nucleus tractus solitarii analysed (medial, dorsal, interstitial, gelatinosus and dorsolateral subnuclei). They appeared to comprise several morphological types, but formed mainly asymmetrical synapses, most often with dendrites of varying size, and contained spherical clear vesicles together with fewer dense-cored vesicles. Substance P- and neurokinin A-immunoreactive terminals were fewer in number (9% of all terminals) but similar in appearance, with the immunoreaction restricted to the dense-cored vesicles. Analysis of serial- and double-labelled sections showed a co-existence of substance P and neurokinin A-immunoreactivity in 21% of glutamate-immunoreactive terminals. Immunoreactivity for gamma-aminobutyric acid was found in 33% of all terminals in the nucleus tractus solitarii. These predominantly contained pleomorphic vesicles and formed symmetrical synapses on dendrites and somata. Possible sites of axo-axonic contact by gamma-aminobutyric acid-immunoreactive terminals onto glutamate-or tachykinin-immunoreactive terminals were rare, but examples of adjacent glutamate and gamma-aminobutyric acid-immunoreactive terminals synapsing on the same dendritic profile were frequent. These results provide an anatomical basis for a gamma-aminobutyric acid mediated inhibition of glutamatergic excitatory inputs to the nucleus tractus solitarii at a post-synaptic level.

Animals↗
PubMed1996

Distribution of hyaluronan in articular cartilage as probed by a biotinylated binding region of aggrecan.

The proportion of total tissue hyaluronan involved in interactions with aggrecan and link protein was estimated from extracts of canine knee articular cartilages using a biotinylated hyaluronan binding region-link protein complex (bHABC) of proteoglycan aggregate as a probe in an ELISA-like assay. Microscopic sections were stained with bHABC to reveal free hyaluronan in various sites and zones of the cartilages. Articular cartilage, cut into 20 microns-thick sections, was extracted with 4 M guanidinium chloride (GuCl). Aliquots of the extract (after removing GuCl) were assayed for hyaluronan, before and after papain digestion. The GuCl extraction residues were analyzed after solubilization by papain. It was found that 47-51% of total hyaluronan remained in the GuCl extraction residue, in contrast to the 8-15% of total proteoglycans. Analysis of the extract revealed that 24-50% of its hyaluronan was directly detectable with the probe, while 50-76% became available only after protease digestion. The extracellular matrix in cartilage sections was stained with the bHABC probe only in the superficial zone and the periphery of the articular surfaces, both sites known to have a relatively low proteoglycan concentration. Trypsin pretreatment of the sections enhanced the staining of the intermediate and deep zones, presumably by removing the steric obstruction caused by the chondroitin sulfate binding region of aggrecans. Enhanced matrix staining in these zones was also obtained by a limited digestion with chondroitinase ABC. The results indicate that a part of cartilage hyaluronan is free from endogenous binding proteins, such as aggrecan and link protein, but that the chondroitin sulfate-rich region of aggrecan inhibits its probing in intact tissue sections. Therefore, hyaluronan staining was more intense in cartilage areas with lower aggrecan content. A large proportion of hyaluronan resists GuCl extraction, even from 20-micrograms-thick tissue sections.

Aggrecans↗