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PubMed1987

Estrogen receptor localization in formalin-fixed, paraffin-embedded endometrium and endometriotic tissues.

Monoclonal estrogen receptor (ER) antibody and indirect immunoperoxidase techniques were used to study the immunohistochemical localization of ER in paraffin sections of 20 surgical specimens containing foci of endometriosis. The eutopic endometrium, available in 16 of 20 cases, was dated histologically and stained for ER. Specific nuclear staining was observed in glandular and stromal cells of the endometrium during the proliferative and early secretory phases. Mid and late secretory phase endometria showed only focal staining. Dating of the endometriotic tissues using morphologic criteria revealed that these were in phase with the endometrium in nine of 16 cases. In these cases the ER staining of both the endometriotic tissues and endometrium was similar. The endometriotic tissues could not be dated as either proliferative or secretory in seven cases and stained variably for ER. Diffuse nuclear staining for ER was present in the endometriotic glands of three of these cases in which the endometriotic glands morphologically resembled glands of the basalis contrasting with weak, focal staining in the corresponding eutopic secretory endometria. The majority of endometriotic tissues examined mimicked the cyclic ER expression of the eutopic endometrium and thus appeared to correspond to the morphologic appearance of the endometriotic glands. Immunohistochemical staining with monoclonal estrophilin antibodies may be helpful in supporting the histopathologic diagnosis of endometriosis.

Adult↗
PubMed1982

Double labeled-antigen method for demonstration of intracellular antigens in paraffin-embedded tissues.

A double "labeled-antigen" method has been developed for the simultaneous staining of both kappa and lambda light chains in fixed paraffin sections. The method is a two step procedure utilizing a mixture of antisera against kappa and lambda light chains in the first stage, followed by the addition of a mixture of kappa antigen labeled with horseradish peroxidase and lambda antigen labeled with alkaline phosphatase. The selection of substrates yielding reaction products of contrasting color enabled the observer to distinguish kappa-containing cells (brown) from lambda-containing cells (blue). Reactive plasma cells stained either pure brown (kappa) or clear blue (lambda) in a ratio of 1.5:1. Blood vessels containing serum immunoglobulins showed a mixed brown-blue reaction, as did the Reed-Sternberg cells of some cases of Hodgkin's disease. The advantages of this double labeled-antigen method over previously reported methods for achieving double staining are discussed.

Antigens↗
PubMed1992

Polymerase chain reaction detects hepatitis B virus DNA in paraffin-embedded liver tissue from patients sero- and histo-negative for active hepatitis B.

The polymerase chain reaction (PCR) was used to analyse tissues from paraffin blocks of liver needle biopsies retrospectively. Biopsies of 29 patients with proven HBsAg and HBcAg expression in liver tissue and of 8 healthy volunteers served as positive (group 1) and negative tissue controls (group 2), respectively. These were compared with 16 patients with proven HBsAg expression in liver but lack of HBcAg (group 3), with 23 patients with anti-HBc as the only hepatitis B virus (HBV)-related marker (group 4) and with 21 patients with liver disease and without HBV markers in tissue or serum (group 5). PCR detected HBV sequences in all cases of the positive control group and in 94% of group 3, in 65% of group 4, and in 71.4% of group 5, whereas all healthy volunteers were negative. Our data show that PCR is able to detect HBV-DNA sequences in virtually all patients with active viral antigen expression but also in a high proportion of hepatitic patients who are silent for active HB but may or may not show signs of a contact with the HBV. Thus, PCR for HBV-DNA in paraffin sections might become a useful tool for identifying patients carrying HBV-DNA but not expressing HBV antigens.

Base Sequence↗
PubMed1976

[An improved histofluorescence procedure for freeze-dried paraffin-embedded tissue based on combined formaldehyde-glyoxylic acid perfusion with high magnesium content and acid pH].

A technique is described for highly sensitive and precise visualization of central catecholamine systems in paraffin sections of freeze-dried tissue. The procedure is based on perfusion of the animal with a solution containing formaldehyde and/or glyoxylic acid, in the presence of a very high magnesium content (40 g MgSO4/150 ml solution) and acid pH. The perfused tissue is rapidly frozen, freeze-dried, treated with formaldehyde vapours (at +80 degrees C for 1 h), embedded in parffin in vacuo, and finally sectioned. The present technique has a sensitivity for the dopamine- and noradrenaline-containing systems that is comparable with that of the glyoxylic acid-Vibratome technique, which utilizes fresh, glyoxylic acid-perfused tissue. Thus, the preterminal axon pathways become fluorescent throughout their full extent and the several new terminal systems, discovered with the glyoxylic acid-Vibratome method, are well demonstrable. The method is also highly useful for the study of the cell bodies and their dendritic processes. The catacholamine fibre systems are visualized without any signs of diffusion and with a richness in detail. In animals pretreated with L-tryptophan and MAO-inhibitor the technique is also useful for studies on central indolamine-containing systems.

Animals↗
PubMed1977

Immunofluorescence studies on formalin-fixed and paraffin-embedded material.

In an attempt to apply immunomorphologic techniques to formalin-fixed material, various tissues were studied by indirect immunofluorescence. To minimize nonspecific background staining, formalin-fixed paraffin sections were treated with pronase prior to incubation with the antisera. This technique was published by Huang in 1975 to study hepatitis B core and surface antigen in formalin-fixed liver-tissue. Our examination demonstrate that this technique might enable the morphologist to identify immunoglobulins in formalin-fixed tissues and so to get new criteria for their diagnosis.

Adult↗