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At least 811 records · Page 45Linked to original sources
PubMed1982

Electron microscopic localization of specific carbohydrate groups in thin sections of tissues embedded in a hydrophilic resin: concanavalin A receptors in mouse spleen.

An improved postembedding method for electron microscopic localization of concanavalin A (Con A) receptors in ultrathin sections is reported. Materials are embedded in a hydrophilic resin, glycol methacrylate copolymerized with glutaraldehyde and urea, using a precisely controlled polymerization schedule that preserves ultrastructural integrity. Ferritin-Con A binds specifically in ultrathin sections of mouse spleen tissue and of Sephadex beads embedded in this resin. Quantitative investigations of procedures for decreasing nonspecific labeling demonstrate that preincubation of sections with bovine serum albumin reduces background to a very low level. This high-resolution method allows macromolecular affinity probes access to receptors in all cellular locations, while maintaining good morphological preservation.

Animals↗
PubMed1988

A simple two-dye basic stain facilitating recognition of mitosis in plastic embedded tissue sections.

Semithin sections of buccal and palatal mucosa fixed in 2.5% glutaraldehyde followed by 1% osmium and embedded in Durcupan (an araldite-based resin) were stained with 2% malachite green in 50% ethanol at 80 C and poststained in 0.05% crystal violet in Sorensen's phosphate buffer (pH 6.4) at 45 C. Nuclear envelopes and chromatin stain vivid purple in contrast to the surrounding green cytoplasm and cell borders. Chromosomes of dividing cells stain bluish violet. Nucleoli, depending on their level in the epithelium, stain differing shades of greenish blue. The distinct and differential staining of each of these components facilitates recognition of mitoses in oral epithelium, where the small size and crowding of cells in the proliferative compartment renders more conventional stains for plastic sections inadequate.

Animals↗
PubMed1993

DNA flow cytometry from paraffin-embedded tissue in diagnosis of cutaneous malignant lymphomas and pseudolymphomas.

DNA content and cell cycle distributions in paraffin-embedded blocks of 111 skin biopsy specimens of 70 patients with cutaneous malignant lymphomas (CML) and 41 patients with cutaneous pseudolymphomas (CPL) including chronic actinic dermatitis (CAD) were estimated by DNA flow cytometry. A statistical significant difference between DNA indices (DIs) or proliferative indices (PIs) for CML including mycosis fungoides (MF) II, MF III, Sezary's syndrome (SS), cutaneous peripheral T-cell lymphomas other than MF and SS, cutaneous germinal center cell-derived lymphomas and cutaneous genuine histiocytic lymphoma from CPL. DIs were also helpful in differentiating MF I from CPL. There was a linear relationship between DIs and PIs, both of which had a parallel relationship to the degrees of malignancy and mortality of varieties of CML. The finding of aneuploidy is likely to be useful in differentiating CML from CPL. It is worth noting that DIs, PIs and proportions of aneuploidy in CAD were all higher than those of higher malignancy of CML. These data could not be considered as markers of malignancy.

Adolescent↗
PubMed1994

Determination of hepatic iron concentration in fresh and paraffin-embedded tissue: diagnostic implications.

BACKGROUND/AIMS: Determination of hepatic iron concentration (HIC) is essential for the evaluation of hereditary hemochromatosis. Occasionally, only paraffin-embedded liver biopsy specimens are available, or fresh biopsy specimens have been placed in saline for transport. This study aimed to describe a method for extraction of liver tissue from paraffin blocks, determine the accuracy of measurement of HIC in recovered tissue compared with fresh tissue, and determine the effect of immersion in saline on HIC. METHODS: HIC was measured in both fresh and deparaffinized liver specimens (n = 41). Accurate measurements were defined as either a normal result in both specimens or a result in the deparaffinized specimen that was within 30% of the fresh measurement. RESULTS: Measurements of HIC in fresh and deparaffinized tissue showed an excellent linear relationship (r = 0.95). In deparaffinized samples > or = 0.4 mg, accurate measurements were seen in 24 out of 29 specimens, compared with 6 out of 12 specimens weighing < 0.4 mg (P < 0.01). The hepatic iron index calculated from results in deparaffinized samples > or = 0.4 mg correctly classified all patients. Immersion of fresh biopsy specimens in saline for 1 hour resulted in up to 50% iron loss (P < 0.05). CONCLUSIONS: Accurate measurement of HIC in deparaffinized liver biopsy specimens is possible. Calculation of the hepatic iron index from deparaffinized liver tissue can facilitate diagnosis of hemochromatosis when fresh tissue is not available. Samples should not be transported in saline.

Animals↗
PubMed1997

A monoclonal antibody pool for routine immunohistochemical detection of human respiratory syncytial virus antigens in formalin-fixed, paraffin-embedded tissue.

Four monoclonal antibodies (MAbs) with specificities for epitopes on human respiratory syncytial virus (RSV) proteins preserved after formalin fixation and paraffin embedding were identified in fixed and embedded virus-infected HEp-2 cell pellets. The MAbs bound epitopes on the fusion protein, the nucleoprotein, the phosphoprotein, and the M2 protein of the virus. Following high-temperature antigen unmasking, immunohistochemical staining revealed RSV antigens in the lungs of five of seven children who died with confirmed RSV infection and in none of nine children who died for other reasons, with no evidence of RSV infection. Staining was cytoplasmic, granular, and confined to epithelial cells. Intense staining was seen at the apex of ciliated bronchial and bronchiolar epithelial cells in all five positive cases. In one case, of pneumonitis, infected pneumocytes were present in the alveoli and in several cases, CD68-positive, cytokeratin-negative alveolar macrophages stained for viral antigens. These antibodies may prove useful in studies of the pathogenesis of RSV infection.

Antibodies, Monoclonal↗
PubMed1993

In situ detection of human Ig light-chain mRNA on formalin-fixed and paraffin-embedded tissue sections using digoxigenin-labelled RNA probes.

Digoxigenin-labelled RNA probes complementary to human immunoglobulin (Ig) kappa and lambda light-chain mRNAs were produced by in vitro transcription. Using these probes, several existing in situ hybridization protocols were studied. By modifying and optimizing pretreatment procedures, which include hybridization, stringency washings and probe detection, a simplified non-radioactive in situ hybridization method for Ig light-chain mRNAs was developed. The light-chain signals were consistently identified in plasma cells, germinal centrocytes, centroblasts and immunoblasts in formalin-fixed and paraffin-embedded sections of lymphoid tissues. Monotypic light-chain mRNA was demonstrated in archival cases of kappa or lambda light-chain-restricted B-cell lymphoma. Background staining was found to be negligible in all the tissues tested. These results indicate that the in situ hybridization methodology described in this study is specific and sensitive for the detection of Ig light-chain mRNAs and has practical value in routine histology.

Antisense Elements (Genetics)↗
PubMed1984

Localisation of enteropathogens in paraffin embedded tissue by immunoperoxidase.

An indirect immunoperoxidase technique has been used to identify enteropathogens in formol-sublimate fixed paraffin embedded sections of calf intestine. Infections with bovine rotavirus, bovine coronavirus, Newbury agent SRV -1, and K99+ Escherichia coli have been detected in the intestines from experimentally infected and conventially reared diarrhoeic or normal calves. The ability to visualize enteropathogenic agents in histological sections resulted in the demonstration of virus infected cells at sites not previously shown to be infected using the immunofluorescence technique.

Animals↗
PubMed1994

Microwave processing for immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded tissue.

The effect of microwave irradiation on the immunoreactivity of proliferating cell nuclear antigen (PCNA) in five primary breast carcinomas, a tonsil with hyperplasia, and a seminoma of the testis was evaluated in formalin-fixed and paraffin-embedded sections using the monoclonal antibody PC-10. The tissues were fixed in 4% buffered formaldehyde for 6, 24, 28, 72 h, and 1 week. Before incubation with the primary antibody, sections were microwaved for x2 5 min in distilled water, in citrate buffer, or they were processed routinely without microwave irradiation. In sections microwaved in distilled water the immunoreactivity was found to be independent of the length of fixation. PCNA immunoreactivity in sections microwaved twice in citrate buffer was not satisfactory. The PCNA immunoreactivity in sections which were routinely processed without microwave irradiation was markedly decreased after 48 h of fixation, and was nearly absent in sections fixed for up to 1 week. In sections processed without microwave irradiation the optimal dilution of the primary antibody was 1:10, whereas in sections microwaved twice in distilled water the optimal dilution was 1:600. The morphology of the tissue was well preserved after microwave processing. We therefore recommend microwave processing in distilled water for x2 5 min for demonstration of PCNA in formalin-fixed and paraffin-embedded sections using the monoclonal antibody PC-10, since this technique yields optimal staining reactions independent of the length of fixation.

Breast Neoplasms↗
PubMed1989

Lymphocyte membrane antigens in glycol methacrylate embedded tissue.

The use of formalin or Michel's solution either alone or in combination with acetone, and acetone, methanol or ethanol alone as fixatives, and glycol methacrylate as embedding medium were evaluated for their suitability in procedures to detect lymphocyte membrane antigens by OKT and Leu monoclonal antibodies in human tonsils. No staining was detected in sections fixed in 70% or absolute ethanol and embedded in glycol methacrylate with either the direct immunofluorescence or avidin-biotin methods. Fixation in Michel's solutions plus acetone at room temperature revealed staining by both. Neither method resulted in staining after fixation in Michel's solution plus acetone at 4 C presumably due to the slow action of the fixative. Staining was enhanced using a combination of primary and secondary biotinylated antibodies. Dual staining allowed concurrent detection of two antigens in the same section. Glycol methacrylate embedding is a possible replacement for ultracold storage in the preservation of tissue for immunofluorescent staining.

Acrylates↗
PubMed1978

Demonstration of the prostatic origin of metastases: an immunohistochemical method for formalin-fixed embedded tissue.

An indirect immunohistochemical technique is described for identification of the prostatic origin of metastases in formalin fixed, paraffin or paraplast embedded material. A rabbit antiserum against the prostate specific acid phosphatase isoenzyme was developed. The method is applicable with or without previous decalcification. In 30 cases of prostatic carcinoma there was only one negative result, and in 20 cases of metastases from prostatic carcinoma positive results were obtained in every instance. All carcinomas (primary focus or metastasis) of non prostatic origin (55) stained negatively with the developed antiserum. The application and possible limitations of the method are discussed.

Acid Phosphatase↗
PubMed1990

Malignant pleural mesothelioma and epidermal growth factor receptor (EGF-R). Relationship of EGF-R with histology and survival using fixed paraffin embedded tissue and the F4, monoclonal antibody.

The expression of epidermal growth factor receptor (EGF-R) in 34 formalin fixed paraffin embedded specimens of malignant mesothelioma was examined using the F4 antibody. Eight samples of reactive pleura showed homogenous cytoplasmic staining with the antibody. EGF-R positive cells (greater than or equal to 5%) were found in 68% of the mesotheliomas examined. EGF-R positivity was more commonly seen in the epithelial histological subtype than in the sarcomatous or mixed subtypes. Patients with less than 5% of mesothelioma cells staining positive for EGF-R had a significantly shorter survival (median 299 days) compared with patients whose tumours had a greater number of cells positive for EGF-R (median 446 days) (P = 0.04). However, when the histological subgroup was also taken into consideration (epithelial type had a significantly longer survival than the sarcomatous or mixed) the survival difference in relation to EGF-R positivity was no longer significant (P = 0.08). EGF-R could not be used to distinguish between malignant and benign mesothelial tissue and was not an independent prognostic factor for survival.

Adult↗
PubMed1980

A device for serially mounting 0.5-2.0 micrometer sections of 1-2 millimeter epon embedded tissue blocks for light microscopy.

The construction and operation of a simple device for serially mounting small Epon sections for light microscopy are described. This device allows the middle portion of the slide to serve both as a shallow reservoir behind the knife edge for flotation of serially oriented sections, and as a miniature hot plate for the subsequent flattening and adhesion of the sections. The advantages of this device, which gave consistently good results with Epon embedded Argas (Persicargas) arboreus tick tissues, are discussed.

Animals↗
PubMed2003

[Detection of COL1A1/PDGFB fusion transcripts in dermatofibroscoma protuberans by revers transcriptase-polymerase chain reaction using paraffin-embedded tissues].

OBJECTIVE: To detect the COL1A1/PDGFB fusion transcripts and discuss its clinicopathological significance in dermatofibroscoma protuberans. METHODS: Formalin fixed, paraffin-embedded tumor specimens from 12 patients with DFSP were reviewed by light microscope and the expression of COL1A1/PDGFB mRNA resulting from the reciprocal translocation t(17;22) (q22;q13.1) was detected by one-step revers transcriptase-polymerase chain reaction. The following tumor specimens were included as controls: 2 fibrosarcoma, 2 malignant fibrous histocytoma, 3 leiomyosarcoma, 1 dermarofibroma and 1 nerve shealth tumor. RESULTS: The COL1A1/PDGFB fusion transcripts were detected in 8 (67%) of 12 samples from patients with DFSP. Nucleotide sequence analysis using the PCR products confirmed that different regions of the COL1A1 gene, respectively, were fused with of PDGFB gene. No COL1A1/PDGFB fusion transcripts were detected in the control tumors. CONCLUSION: Detection of specific COL1A1/PDGFB fusion transcripts in DFSP will help to diagnose the nature of DFSP and research the mechanism of its molecular histogenesis.

Adolescent↗
PubMed1988

Diagnosis of nasopharyngeal tuberculosis by detection of tuberculostearic acid in formalin fixed, paraffin wax embedded tissue biopsy specimens.

The use of gas chromatography and mass spectrometry with selected ion monitoring detected tuberculostearic acid (TBSA) in 10 of 12 formalin fixed, paraffin wax embedded nasopharyngeal and head and neck biopsy specimens from patients with confirmed tuberculosis and carcinoma, and in one of 50 control specimens (giving a sensitivity of 83% and a specificity of 98%). The two false negative cases had very small tissue fragments and the patient with a false positive result may have had pulmonary tuberculosis. Tuberculostearic acid (TBSA) was also detected in nine of 16 specimens from the head and neck region with non-caseating granulomas suspected, but not confirmed, to be tuberculosis. It is concluded that nasopharyngeal tuberculosis is relatively common in Hong Kong and should be considered when biopsy specimens show granulomas. The detection of TBSA in tissue biopsy specimens is a useful, rapid method for the diagnosis of tuberculosis and other mycobacterial infections, and can be conveniently performed within two days on formalin fixed and paraffin wax embedded material.

Granuloma↗