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 739 records · Page 41Linked to original sources

An attachment tip and pili-like structures in insect- and plant-pathogenic spiroplasmas of the class Mollicutes.

Ultrastructural studies using scanning electron microscopy (SEM), negative-staining transmission electron microscopy (TEM), and thin-sectioning TEM on four species of Spiroplasma, in vitro and/or in vivo, indicated that their helices commonly possess one tapered end (tip structure) and one blunt or round end. These tip structures appeared morphologically different from the rest of the helix, exhibiting an electron-dense conical or rod-shaped core. In thin sections of the midgut of the leafhopper Dalbulus elimatus, the tip structures of Spiroplasma kunkelii in the midgut lumen were mostly aligned between microvilli, perpendicular to the apical plasma membrane of epithelial cells. These tip structures appeared frequently attached or closely apposed to the plasma membrane, in which cup-shaped invaginations close to the tips were observed. Pleomorphic forms of spiroplasma, enclosed in membranous vesicles, were found in the cytoplasm of the midgut epithelial cells. These findings suggest that the tip structure may be involved in the orientation and attachment of spiroplasma helices in relation to their host cells, and thus may be functionally comparable to the "attachment organelle" of mycoplasmas. Additionally, pili-like structures were observed by negative-staining TEM on the surface of Spiroplasma melliferum, and in thin sections of S. kunkelii infecting the leafhopper vector Dalbulus gelbus.

Animals↗

Glycine- and GABA-immunoreactive nerve terminals apposed to alpha-motoneurons during postnatal development in kittens: a quantitative study using the disector.

The distribution of gamma-aminobutyric acid (GABA) and/or glycine-immunoreactive (IR) terminal-like structures apposed to somatic and dendritic membrane of lumbar alpha-motoneurons in column 2 was examined in 1- and 3-week-old kittens and in the adult cat. This quantitative study was carried out using a postembedding technique on semithin sections and a stereological method, the disector. Analysis of immunoreactive terminals showed that the percentages of GABA-IR and glycine-IR terminals (these populations include terminals containing both GABA and glycine) apposed to somatic and proximal dendritic compartments of alpha-motoneurons are almost the same in kittens, while in the adult glycinergic innervation becomes predominant. This change results from: (1) the decrease in numbers of GABA-IR terminals contacting the somatic compartment between 3 weeks and adult stage, while the numbers of glycine-IR terminals show no significant changes after birth and the numbers of terminals containing both neurotransmitters (GABA-IR+glycine-IR) present transient changes and (2) the postnatal increase in the dendritic compartment, in numbers of GABA-IR, glycine-IR and GABA-IR+glycine-IR terminals; the increase being larger for glycine-IR terminals. Furthermore, using a postembedding immunogold technique, observations by electron microscopy showed that GABA-IR P boutons apposed to M boutons can already be identified at 1 week after birth.

Age Factors↗

Villus and crypt cell composition in the secreting mouse jejunum measured with x-ray microanalysis.

The response of the villus and crypt cells of the mouse jejunum to secretagogues has been assessed through measurements of cellular composition with x-ray microanalysis. In nonstimulated tissues the Na concentration ([Na]c) of the crypt cells was significantly less, and the K ([K]c) and Cl ([Cl]c) concentrations were significantly greater, than that of the villus cells. There was also a decreasing gradient of [Na]c and increasing gradient of [K]c from the villus tip to crypt base due to a greater number of cells with a high [Na]c and low [K]c in the upper regions of the villi. Theophylline (10 mmol L-1) stimulated a sustained increase in bumetanide sensitive short circuit current (Isc) and significantly decreased the [Na]c of the villus cells. Similar, but smaller changes were seen in the crypt cells. Changes in villus cell [Na]c reflected a reduction in the number of cells with a high [Na]c. Inhibition of the apical Na/H exchanger (1 mmol L-1 amiloride) had little effect on basal Isc and the subsequent addition of theophylline increased Isc to a comparable extent as seen without amiloride. However, after amiloride treatment the only change in cellular composition was a reduction in the [Cl]c of both crypt and villus cells, suggesting that both regions are involved in the secretory response. These data suggest that the dominant response of the jejunum to secretagogues is an inhibition of Na absorption via Na/H exchange in the villi and the secretory response is distributed throughout the crypt/villus axis.

Amiloride↗

Histological study of the human temporo-mandibular joint and its surrounding muscles.

This is a histological study of the human temporo-mandibular joint and its surrounding muscles. Using a microscopic study of serial sections from anatomical specimens from six subjects, the detailed anatomy of the joint is presented with particular regard to the histology. This study has allowed, in particular, the description of the ligaments and capsule as well as the insertions of the masticatory muscles (temporalis, masseter, lateral pterygoid) on this joint. These observations are then compared with the anatomical and histological data already reported on this subject. This study shows that the bulk of the muscular fibres of the lateral pterygoid passes under the foot of the disc is attached over the whole height of the condylar, unite and extend as far as the medial pole of the joint under the insertion of the articular disc. An insertion of the temporo-masseter musculo-tendinous complex on the anterior and lateral capsulo-discal structures was observed. The lateral pterygoid is composed of a succession of tendinous and fleshy fibres. This study confirms the thickening of the lateral capsule that corresponds to a lateral collateral ligament, and the absence of a medial collateral ligament. Medial stability is conferred by the lateral ligament of the contralateral joint.

Aged↗

Potential of P40 plastination for morphometric hip measurements.

Total hip replacement has become one of the most successful surgical operations over the past 25 years. The duration of a total hip prosthesis depends on primary stability, and many studies have tried precisely to evaluate hip joint morphology to obtain excellent contact between bone and prosthetic component. This study performed a morphometric analysis of the human hip joint using, for the first time, the P40 plastination procedure. We cut 42 hip joint compounds into slices 3 mm thick; for exact distance measuring the sections were scanned into the computer. The following mean measurements for hip geometry were obtained: vertical diameter of acetabulum 4.894+/-0.274 cm, depth of acetabulum 1.643+/-0.245 cm, femoral head radius 2.268+/-0.149 cm, femoral neck length 4.3670+/-0.528 cm, acetabular perimeter 6.711+/-0.434 cm, vertical diameter of labrum acetabulare 4.759+/-0.476 cm, depth of labrum acetabulare 2.599+/-0.395 cm, sum of femoral head and neck lengths 6.759+/-0.550 cm, hip axis length 11.859+/-1.007 cm, femoral neck axis length 10.12+/-0.555 cm, and femoral neck diameter 3.349+/-0.276 cm. All of these data reveal a significant gender difference. Our aim was to indicate an unconventional and new method of gaining morphometric hip data by using plastination.

Acetabulum↗

Binswanger's encephalopathy: serial sections and morphometry of the cerebral arteries.

To identify arterial changes that are characteristic of Binswanger's encephalopathy (BE), we analyzed cerebral subarachnoid and medullary arteries of seven BE autopsy specimens by reconstruction of stained serial sections. We also noted the frequency of intimal fibrosis with or without atheroma of the subarachnoid arteries, and determined the medial thickness of the subarachnoid and medullary arteries. The results for the BE specimens were compared with those of six hypertensive brain hemorrhage (HH) specimens and six normotensive (NT) specimens from patients without cerebral abnormalities. In medullary arteries of BE in comparison with HH, we observed nonspecific but significantly more widespread intimal fibrosis with or without atheroma, as well as segmental loss of the medial smooth muscle cells (SMCs), which was sometimes associated with intimal plasma exudation or microaneurysm. A few medullary arteries in BE were completely occluded by fibrous connective tissue. Intimal fibrosis of the subarachnoid arteries was significantly more widespread in BE than in HH and NT. The media of the subarachnoid and medullary arteries was significantly thicker in BE and HH than in NT, and tended to be thicker in BE than in HH. In NT specimens the medullary arteries tended to be thinner in medial thickness than the subarachnoid arteries. These findings suggest that dysfunction of blood flow regulation due to increased arterial stiffness caused by hypertension-induced intimal fibrosis and loss of medial SMCs is an essential mechanism resulting in diffuse myelin loss of the cerebral white matter in BE, whereas luminal stenosis or occlusion and adventitial fibrosis are secondary. Moreover, selective and severe involvement of the cerebral medullary arteries compared with the subarachnoid arteries may be explained by the following two factors, (1) that many medullary arteries have normally dilated segments, and (2) that their media is thinner compared with that of the subarachnoid arteries of the corresponding diameter.

Aged↗

Loss of the TSC2 product tuberin in subependymal giant-cell tumors.

We investigated immunocytochemically the expression of tuberin, the TSC2 gene product, in brain resections from children with and without tuberous sclerosis, to characterize the phenotype of balloon and tumor cells, and to elucidate the relationship between tuberin and formation of subependymal giant-cell tumors. In cortical tubers, tuberin was expressed in processes and cell bodies of balloon cells, which also showed consistent vimentin and nestin immunoreactivity, but no glial fibrillary acidic protein, neurofilament, or galactocerebroside positivity by immunofluorescence confocal microscopy. The majority of balloon cells in white matter showed weaker tuberin immunoreactivity than similar cells in cortical tubers. In subependymal giant-cell tumors, there was minimal to no immunoreactivity. Why tuberin is expressed in tubers but not in subependymal giant-cell tumors is not known. If tuberin plays a role in tumor suppression, then lack of tuberin might be a marker for deletion of both alleles, leading to enhanced cellular growth in subependymal giant-cell hamartomas, eventually of sufficient size to cause clinical symptoms.

Adolescent↗

Use of electron spectroscopic imaging to determine element composition of the melanin granules in the stria vascularis of the guinea pig.

Electron spectroscopic imaging (ESI) was used to analyze the element content of melanin granules in the stria vascularis seen in ultrathin sections of Spurr-embedded cochleae of the guinea pig. To determine element composition, ESI images were taken at different ionization edges, and non-specific background signals were subtracted digitally by an image processing system. The presence of calcium and nitrogen in the melanin granules could be demonstrated clearly. The calcium identified in the melanin granules was then compared with the spatial distributions of calcium binding sites after the application of an antimonate precipitation method, which was used to localize loosely bound calcium. Despite a high calcium concentration within the granules, only very small single scattered calcium precipitates could be detected between these structures as compared with the amount of calcium precipitates attached to the plasma membrane or located within the cell nuclei. The nearly complete absence of precipitates within the melanin granules after the application of antimonate suggests differences in calcium binding and mobility involved in various physiological processes of ion balance regulation within the stria vascularis.

Animals↗

The muscular tissue of the vestibular folds of the larynx.

The muscular tissue of the vestibular folds was investigated in plastinated serial sections of 32 normal adult larynges. Three muscular systems could be distinguished. A posterolateral muscle layer was found to be developed at the lateral margin of the posterior part of the vestibular fold. Its fibers extended in a sagittal direction, and their contraction probably resulted in an adduction of the entire tissue of the vestibular fold towards the midline. Within the anterior part of the vestibular fold, an anterolateral muscle sheet was seen to attach to the thyroid cartilage. An anteromedial muscular system consisted of scattered groups of muscle fibers situated medially and dorsally to the laryngeal ventricle and saccule. These fibers were presumed to exert a downward pressure on the vestibular folds, in addition to an adductor function. According to clinical experience, adductor movements of the vestibular folds can be trained, even in cases with a recurrent laryngeal nerve lesion, in order to produce a compensatory voice. Thus, the muscles of the vestibular folds are probably innervated by the superior laryngeal nerve.

Adult↗

An optimized method for simultaneous demonstration of neurons and myelinated fiber tracts for delineation of individual trunco- and palliothalamic nuclei in the mammalian brain.

The truncothalamic complex has long been considered to be a nuclear group with "non-specific" projections. More recently, it is suggested that these thalamic nuclei play an important role in regulating distinct basal ganglia circuits. To further analyze the exact biological function of individual nuclei of the truncothalamic complex a simple and reliable technique for an exact delineation of distinct nuclei is desirable. Therefore, we evaluated and optimized several potential procedures for a combined visualization of neurons and myelinated fibers. Fiber staining with gold toning or immunocytochemical visualization of myelin basic protein shows high contrast and precision but precludes sufficient demonstration of neuronal cell bodies. When the most common technique for the simultaneous visualization of both structures, the Kluver-Barrera procedure, is used, demonstration of myelinated fibers is restricted when the technique is applied to cryostat or vibratome sections. In the present report this limitation was abolished. The final protocol includes lipid extraction prior to the incubation with Luxol Fast Blue and uses carefully characterized staining conditions for Luxol Fast Blue and cresyl violet rendering microscopically controlled differentiation steps unnecessary. The optimized Kluver-Barrera technique results in high precision localization of individual axons and cell bodies and thus permits an exact and simple delineation of individual nuclei in the vertebrate thalamus.

Animals↗

Immunofluorescence technique for 100-nm-thick semithin sections of Epon-embedded tissues.

An immunofluorescence staining method for Epon-embedded materials is described. Rat kidney and liver were fixed by perfusion with 1% glutaraldehyde for 10 min. Tissue slices were embedded in Epon. Semithin sections with thicknesses ranging from 1,000 to 100 nm were cut and mounted on clean glass slides. Epoxy resin was removed by treatment with 10% sodium ethoxide. Sections were digested with 0.05% trypsin and then treated with sodium borohydride. Sections were immunostained for leucine aminopeptidase (plasma membrane), catalase (peroxisomes), 3-ketoacyl-CoA thiolase (mitochondria), cathepsin D (lysosomes), and LGP107 (lysosomal membrane) using Cy(2)- or Alexa 546-labeled secondary antibodies. In 1,000-nm-thick sections, non-specific fluorescence remained and such fluorescence decreased as the sections became thinner. Clear specific fluorescence was obtained in the sections with thicknesses ranging from 250 to 100 nm with Alexa 546-labeled antibody (red fluorescence) but was not specific enough with Cy(2)- or Alexa 430-labeled antibody (green fluorescence). Sodium borohydride greatly abolished autofluorescence of glutaraldehyde. The present method made it possible to obtain signals in cross-sections of biological materials with a thickness of 250-100 nm, which are difficult to obtain in optical section using a confocal laser microscope.

Acetyl-CoA C-Acyltransferase↗

A method for preparing 2- to 50-micron-thick fresh-frozen sections of large samples and undecalcified hard tissues.

This article describes a method for preparing 2- to 50-micron-thick fresh-frozen sections from large samples and completely calcified tissue samples. In order to perform the more routine work involved, a tungsten carbide disposable blade was installed to a heavy-duty sledge cryomicrotome. An entire 10-day-old rat and bone and tooth samples from a 7-month-old rat were rapidly frozen. The frozen samples were attached to the cryomicrotome stage. The cutting surface of the samples was covered with a polyvinylidene chloride film coated with synthetic rubber cement and cut at -25 degrees C. The soft tissues and the hard tissues were satisfactorily preserved and all tissue cells were easily identifiable. Enzymatic activity in the fresh sections was much stronger than that in chemically fixed and/or decalcified sections. The sections permitted histological and histochemical studies without trouble. In addition, the sections can be used for multiple experiments such as immunohistochemistry, in situ hybridization, and electron microprobe X-ray micro-analysis. This method can be used with conventional cryomicrotome equipment.

Alkaline Phosphatase↗

Signal enhancement at the electron microscopic level using Nanogold and gold-based autometallography.

Immunoelectron microscopy using ultrasmall gold markers is a very sensitive method to detect molecules at high resolution. In order to discriminate the gold particles in the electron microscope, enlargement of gold particles is necessary. So far, mostly silver ions were used for deposition onto the surface of gold grains. In our study, we tested the selective deposition of gold instead of silver ions to enlarge gold particles. This was performed following immunogold detection of DNA at the surface of ultrathin sections embedded in the acrylic resin LR White (postembedding approach). Morphometric analysis of the distribution of DNA in human spermatocytes revealed that the method offers very good specificity and sensitivity and therefore is a good alternative to the use of silver for signal enhancement. This technique was also applied to the detection of ribosomal genes in human testis at the electron microscopic level by in situ hybridization. Ribosomal genes were localized in peri- and intranucleolar chromatin as well as in the dense fibrillar component of nucleoli.

Cell Nucleolus↗

Multi-colour brightfield in situ hybridisation on tissue sections.

We describe the brightfield microscopical detection of multiple DNA target sequences in cell and tissue preparations. For this purpose, chromosome-specific DNA probes labelled with biotin, digoxigenin or fluorescein were simultaneously hybridised and detected by enzyme cytochemistry using two horseradish peroxidase (PO) reactions and one alkaline phosphatase (APase) reaction. For triple-colour detection on single cell preparations, the combination of the enzyme precipitates PO/diaminobenzidine (DAB, brown colour), APase/fast red (FR, red colour) and PO/tetramethylbenzidine (TMB, green colour) resulted in an accurate detection of DNA targets. Embedding of the preparations in a thin cross-linked protein layer further stabilised the enzyme reaction products. For in situ hybridisation on tissue sections, however, this detection procedure showed some limitations with respect to both the stability of the APase/FR and PO/TMB precipitates, and the sequence of immunochemical layers in multiple-target procedures. For this reason, the APase/FR reaction was replaced by the APase/new fuchsin (NF, red colour) reaction and the washing steps after the PO/TMB reaction were restricted to the use of phosphate buffer pH 6.0. Furthermore, to improve the efficiency of the ISH reaction, APase/NF was applied in an avidin-biotin complex detection system and, to avoid target shielding in the triple-target ISH, the third primary antibody was applied prior to the second enzyme cytochemical reaction. These adaptations resulted in stable, well contrasting brown, red and green coloured precipitates. After quick haematoxylin counterstaining, the tissue preparations were directly mounted in phosphate buffer and, optionally, embedded in the cross-linked protein layer.

Color↗

Transmission light microscopy of structurally colored semithin cartilage sections.

Conversion of osmified tracheal cartilage constituents into an array of laminar interference gratings has been attained by three tandem reactions. Oxidation of semithin, LR white-embedded cartilage sections by acetic anhydride in dimethyl sulfoxide is the first step in the conversion process. Subsequent addition reactions of oxidized cartilage pyranoses and furanoses with thiocarbohydrazide constitutes the second step. Reduction of silver proteinate by thiocarbohydrazones and the concomitant coating of cartilage constituents with silver gratings completes the conversion of cartilage sections into a system of layered interference filters. In transmitted light, all components of converted cartilage display vivid structural colors, which allow detailed microscopic analysis of structurally colored cellular and extracellular cartilage constituents.

Animals↗

Identification of target cells by immunohistochemical detection of covalently rearranged estradiol in rehydrated paraffin sections.

Estradiol is released from the binding niche of the receptor and covalently arrested in the molecular vicinity by the Mannich reaction during target fixation in acetic acid/formaldehyde. The exposed steroid is freely accessible for appropriate antibodies. It can be visualized in sections by the second antibody/enzyme technique in high resolution and without enhancements.

Animals↗

Differential expression of IFN-gamma, IL-4, IL-10, and IL-1beta mRNAs in decalcified tissue sections of mouse lipopolysaccharide-induced periodontitis mandibles assessed by in situ hybridization.

To examine the role of T cell subgroups, Th1 and Th2, in the development of periodontitis, the expression of various cytokines was investigated in a mouse model of alveolar bone resorption using in situ hybridization (ISH) with digoxigenin-labeled oligonucleotides. When mice received repetitive injections with Escherichia coli lipopolysaccharide into the gingiva every 48 h, alveolar bone resorption was detectable after the fourth injection, reaching a maximum after the 13th injection. For the best performance of ISH, we first had to choose a decalcification protocol. Among various decalcification protocols, 10% EDTA (4 degrees C, 5-6 days) was the best for 28S rRNA staining. Positive cells for transcripts of interferon-gamma (Th1 product) were detected after the fourth injection, reaching a maximum after the tenth injection. A similar pattern was obtained for interleukin (IL)-10 mRNA (Th2 product) and IL-1beta, while the positive cell number reached a maximum after the 13th and 10th injections, respectively. The number of IL-4 mRNA (Th2 product)-positive cells remained low till the tenth injection, but increased thereafter. Consequently, we found that the population change from Th1 to Th2 in the inflammation site correlated with the transition from gingivitis to periodontitis, indicating differential roles of T cell subgroups in the development of periodontitis.

Alveolar Bone Loss↗

A new radiographic procedure for obtaining correction factors of 3H-beta-self-absorption for quantitative tritium autoradiography.

A 3H-radiographic method based on the absorption of 3H-beta-particles by an overlying tissue section was established for obtaining correction factors of 3H-beta-absorption (c.f.s) on the cellular level for all kinds of sections in a simple and more precise way as possible with interferometry. Unlabelled paraffin or Araldite sections were mounted on a thin uniformly 3H-labelled section of resin, and autoradiographs were prepared. Grain densities of neuronal cell types and cell-free areas within and outside the paraffin or Araldite sections were evaluated in autoradiographs, where the exposure time or the thickness of the overlying histological section was varied. From these values c.f.s were calculated applying the Beer-Lambert law. It was shown that corresponding c.f.s determined with this new radiographic method correspond well with each other. However, they will only agree with those c.f.s obtained by interferometry, if relative c.f.s are compared. Since the c.f.s are quite sensitive to the section thickness, a new parameter phi was introduced, which helps to assess whether the microtome used works exactly. Generally, the method presented can be used on the cytological level as well as for whole areas in every other autoradiographic study.

Animals↗