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At least 631 records · Page 35Linked to original sources
PubMed1984

Identification of thymostimulin secreting cells in calf thymus by immunoperoxidase method.

An anti- thymostimulin (TS) serum was tested on calf thymus to study the localization of the hormonal factor. The immunoperoxidase method was applied to tissue fixed in Bouin's fluid and embedded in paraffin, or to tissue fixed in paraformaldehyde and embedded in Epon for semi-thin sections. Immuno-reactivity was shown, with DAB- Fluka , in reticulo-epithelial cells in the medulla, and between the cortex and the medulla, while with DAB-Sigma reactivity was found in the cortex as well. The external cells of Hassall's corpuscles were also reactive. Myoid cells were not reactive. In semi-thin sections a weak reactivity was noted at the periphery of a few lymphocytes. Comparison with the localization of other thymic factors, and the possibility of a functional cycle of the epithelial cells synthesizing one or more factors are discussed.

Animals↗
PubMed1983

A rinsing and incubation chamber used for immunocytochemistry of vibratome sections.

This paper describes the construction and use of a washing/incubation chamber that may be used for peroxidase-antiperoxidase immunocytochemistry of vibratome sections. The advantages of the chamber include: all sections are processed simultaneously with exception of exposure to the primary antiserum; the exposure of the sections to DAB substrate can be precisely controlled; and also the sections are not directly manipulated during the procedure resulting in a much greater integrity of the tissue. Use of the chamber reduces the amount of labor normally required to wash the sections throughout the procedure. Application of this system to the development and standardization of quantitative immunocytochemical techniques is also discussed.

Immunoenzyme Techniques↗
PubMed1975

Observations on the selective demonstration of nucleolar material by protein staining techniques in epon thick sections.

The effect of different protein stainings on thick sections of Epon embedded salivary glands of Chironomus pallidivittatus is described. Pretreatment of sections with several metallic salts affected the affinity for aniline blue black in different structures. The uranyl and aluminium ions followed by aniline blue black resulted in a highly selective nucleolar staining which proved also efficient in Allium cepa meristematic cells and which seems useful for the preferential demonstration of nucleolar material in thick sections.

Animals↗
PubMed1986

Ultracryotomy for the study of unfixed membranes.

Plasma membranes from chromaffin cells of bovine adrenal medullae and from chicken macrophages were isolated on a urografin density gradient, frozen and sectioned without previous chemical fixation. Their receptor binding sites were localized by specific labelling. The sections were then post-fixed in the presence of K2Cr2O7 to produce positive staining of the membrane proteins. Chromaffin cell membranes formed single vesicles. The nicotinic acetylcholine receptor (localized using a monoclonal antibody against its cholinergic binding site) was always found in patches on the surface of vesicles, whose profiles corresponded to thickened bilayers. Macrophage membrane vesicles were agglutinated. The mannose receptor (localized using the ligand, mannosylferritin) was randomly distributed within the electron-dense coat of the agglutinated vesicles or on electron-dense caps involved in agglutination. The binding sites of both receptors were intact, as revealed by their being recognized by a monoclonal antibody against their cholinergic binding sites and by the active binding of the mannosylated ligand which was inhibited by mannan. The distribution of the receptors on the vesicles reflected their distribution on the cell surface.

Adrenal Medulla↗
PubMed1989

Cryostat sections for coexistence studies and preembedding electron microscopic immunocytochemistry of central and peripheral nervous system tissue.

Perfusion-fixed tissue blocks were incubated in high molar sucrose solutions, shock frozen in melting isopentane, and sectioned on a conventional cryostat. Semithin sections (2-4 microns) alternatingly stained for parvalbumin and glutamate decarboxylase enabled us to demonstrate the coexistence of both antigens in the same cell. Thick sections (40 microns) of central and peripheral nervous system tissue were immunostained and processed for correlated light and electron microscopic studies. At the electron microscopic level, the preservation of ultrastructural features such as membranes and synaptic contacts was comparable to that normally seen in vibratome sectioned material. Hence, this technique can successfully be used for preembedding coexistence studies and electron microscopic preembedding immunocytochemistry when vibratome sectioning is problematic.

Animals↗
PubMed1987

Serial section method for cytofluorometric determinations of the DNA content of component cells of the human cochlea.

We describe a method for measuring the DNA content of the component cells of the organ of Corti using serial sections of human cochleae obtained at autopsy. Cochleae were fixed in Carnoy's solution and embedded in Acrytron E, a water-miscible methacrylate resin. A procedure was developed to reduce the background fluorescence in methacrylate-embedded sections; the resin was pretreated with ion-exchange resin (Amberlite IRA-410). Experiments showed that pretreatment reduce the background fluorescence practically to zero. Seventy 3 microns-thick serial sections were prepared on fluorescence free glass slides and stained with azocarmin G and acriflavine-Feulgen. After postirradiation using blue excitation light, the amount of Feulgen-DNA present in the target nucleus in each section was determined using a microfluorometer. The amount of DNA in the entire nucleus was determined by adding together the DNA content of the segments of the nucleus. The characteristic appearance of the organ of Corti made it easy to detect these cells; under green excitation light the cells of this organ exhibited red cytoplasmic azocarmin-G fluorescence. Due to the relatively wide internuclear spaces, cytofluorometry fo individual nuclei could be performed without interference from the neighboring cells. Our technique using serial sections allowed us to measure the DNA content of individual cells and obtain histological information about particular cells and their neighboring cells. Several polyploid cells were found among the Hensen's cells in the cochlea, while all other component cells of the organ of Corti were diploid.

Cell Nucleus↗
PubMedDate not supplied

Microtome-cryostat.

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Biomedical Engineering↗
PubMed1989

A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system.

(1) A preparation is described which allows patch clamp recordings to be made on mammalian central nervous system (CNS) neurones in situ. (2) A vibrating tissue slicer was used to cut thin slices in which individual neurones could be identified visually. Localized cleaning of cell somata with physiological saline freed the cell membrane, allowing the formation of a high resistance seal between the membrane and the patch pipette. (3) The various configurations of the patch clamp technique were used to demonstrate recording of membrane potential, whole cell currents and single channel currents from neurones and isolated patches. (4) The patch clamp technique was used to record from neurones filled with fluorescent dyes. Staining was achieved by filling cells during recording or by previous retrograde labelling. (5) Thin slice cleaning and patch clamp techniques were shown to be applicable to the spinal cord and almost any brain region and to various species. These techniques are also applicable to animals of a wide variety of postnatal ages, from newborn to adult.

Animals↗
PubMed1975

[Determination of intracapillary HbO2 saturation with a cryo-microphotometric method, applied to the rabbit myocardium (author's transl)].

We succeeded in determining the O2 saturation of Hb in capillaries of the rabbit myocardium with a cryo-microphotometric method. Myocardial samples (2-3 mm) were removed in situ with a pair of nitrogin-cooled copper tongs, and rapidly frozen in Freon 13. The samples were sectioned into 10-12 mum slices in a cryotome at about minus 60 degrees C. Hemoglobin absorption spectra were measured on the capillary sections of the frozen slices at minus 100 degrees C in a vacumm-isolated microscope cooling chamber which we developed for the UMSP 1 (C. Zeiss, Oberkochen). The spectra were subdivided into three basic components (oxygenated Hb, desoxygenated Gb, and desoxygenated dehydrated Hb) with a weighted multi-component analysis (Lübbers and Wodick, 1969). The basic components were measured in Hb solutions. The accuracy of the method was tested both with Hb solutions inserted into glass capillaries, and Hb droplets. O2 saturations of the Hb solutions were measured at room temperature. Subsequently, both test solutions were frozen, HbO2 saturation measured in the glass capillaries at low temperatures agreed well with those recorded at room temperature. HbO2 saturation of the sectioned hemoglobin droplets was found to be systematically increased with low Hb32 saturations as compared with the records made at room temperature. The systematic error was determined and the intracapillary HbO2 saturations were corrected accordingly. Measurements on samples of the rabbit myocardium showed that the histogram of capillary HbO2 saturations has a maximum at saturations of 20-30%. The lowest saturation values were found to be between 0 and 10%, but these values were recorded less frequently. The arterial HbO2 saturation in the femoral artery was about 98%.

Animals↗