Periodate-Lysine-Paraformaldehyde Fixative a New Fixative for Immunoelectron Microscopy

McLean, Ian W.; Nakane, Paul K.

doi:10.1177/22.12.1077

Cited by 3,666 publications

(1,315 citation statements)

References 10 publications

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“…However, even under the EM, the differentiation between macrophages and IDCs has proved difficult in synovial tissue, because the method of fixation used for monoclonal antibody staining (26) obscures the fine definition of the cellular organelles. Nevertheless, in those transitional areas which were relatively rich in lymphocytes (50-70% of the total cell population), 63D3-cells with the morphologic appearance of IDC formed 10-20% of the DR+ macrophage-like cell population.…”

Section: Discussionmentioning

confidence: 99%

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Electron microscopic study of HLA‐DR and monocyte/macrophage staining cells in the rheumatoid synovial membrane

Iguchi

Kurosaka

Ziff

1986

Arthritis & Rheumatism

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We examined synovial membrane samples from 6 rheumatoid arthritis (RA) and 3 osteoarthritis patients and from 1 normal subject, by an immunoelectron microscopic technique using anti-HLA-DR (anti-Ia) and anti-monocyte/macrophage (63D3) monoclonal antibodies. In the lining layer, the type A macrophage-like cells were strongly DR+ and 63D3+, whereas the type B fibroblast-like cells were almost completely negative. Lymphocyte-rich areas (containing more than 90% densely packed lymphocytes) showed weak and patchy DR staining of the lymphocytes. In these areas, 3 4 % of the cells were macrophage-like cells which were 63D3-, a type of staining compatible with that of the interdigitating cell (IDC). In the plasma cell-containing (transitional) areas, many strongly DR+ macrophage-like cells were observed in close contact with lymphocytes and plasma cells. Ten to twenty percent of these cells were 63D3-, which suggests that they were IDC. Cells with the structural appearance of IDC were most frequently seen in those transitional areas which contained elevated concentrations (50-70%) of lymphocytes. In uninfiltratFrom the Harold C.

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Section: Discussionmentioning

confidence: 99%

“…The technique of this procedure has been previously described (26). Briefly, synovial specimens were cut into approximately 4-mm3 pieces and were promptly fixed with paraformaldehyde-lysine-periodate solution (27) for 6 hours.…”

Section: Methodsmentioning

confidence: 99%

Electron microscopic study of HLA‐DR and monocyte/macrophage staining cells in the rheumatoid synovial membrane

Iguchi

Kurosaka

Ziff

1986

Arthritis & Rheumatism

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“…The cells were suspended at a concentration of 107/ml. , and 2% paraformaldehyde (14). The fixed cells were washed twice with PBS to remove any fixative.…”

Section: Materials and Methods Patients And Normal Controlsmentioning

confidence: 99%

“…Developed initially for immunoelectron microscopy, PLP has recently been shown to be a valuable fixative for the improved detection of cell surface antigens, particularly in lymphoid tissue (14,151. Because of the preservation of surface antigens, which was also documented in our study, it is possible to examine surface markers and Ki-67 expression a t the same time.…”

Section: Discuss I 0 Nmentioning

confidence: 99%

Simultaneous flow cytometric analysis of surface markers and nuclear Ki‐67 antigen in leukemia and lymphoma

et al. 1989

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The monoclonal antibody Ki‐67 identifies an antigen present during the late G1, S, G2, and M phases of the cell cycle, whereas resting cells do not express this antigen. Immunostaining with Ki‐67 provides a simple method with which to determine the growth fraction of a malignant cell population without requiring a laborious procedure or use of radioactive materials. Thus far, detection of Ki‐67–positive cells by flow cytometry was limited because of nuclear location of the antigen. In this study, periodate‐lysine‐paraformaldhyde (PLP) fixation of cells in suspension, labeling with Ki‐67, and the subsequent flow cytometric analysis of the tumor growth fraction is described. Fixation with PLP at −10°C for 15 min rendered the plasma membrane permeable without destroying cell surface antigens. Thus double immunofluorescence studies using both a surface marker and Ki‐67 could be performed. This offers the additional advantage of being able to define the phenotype of proliferating cells. This method was applied to determine the growth fraction in peripheral blood and bone marrow samples of patients with leukemia and non‐Hodgkin's lymphoma. The results of Ki‐67 studies in 91 patients are shown. A wide variability of individual Ki‐67 values was observed within each entity. Use of this flow cytometric procedure substantially facilitates the quantification of proliferating cells in pathological blood and bone marrow samples.

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“…To detect the Mac-2 antigen, namely galectin 3 (17,18), tissue samples were fixed with periodate-lysineparaformaldehyde solution (19) overnight at 4°C before freezing. Immunostaining was performed using rat anti-Mac-2 antibodies (Boehringer, Mannheim, Germany), biotinylated rabbit anti-rat IgG antibodies, and then FITC-labeled avidin or avidin-biotin-peroxidase complexes.…”

Section: Methodsmentioning

confidence: 99%

Rheumatic diseases in an MRL strain of mice with a deficit in the functional Fas ligand

Ito

Terasaki

Itoh

et al. 1997

Arthritis & Rheumatism

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Objective. To characterize Fas antigen expression on the cell surface, and to determine the effect of this expression in rheumatic diseases using a newly established gld-congenic MRL strain of mice (MRLlgld), which is defective in its functional Fas ligand (Fas-L).Methods. Flow cytometric analyses of lymphoid cells and macrophages were performed using anti-Fas and other cell surface markers. Histopathologic manifestations were examined using immunochemistry and light and electron microscopy. Serum levels of IgG and anti-DNA antibodies were measured by single radial immunodiffusion and enzyme-linked immunosorbent assay, respectively.Results. MRL/gld mice developed systemic lymphadenopathy with an accumulation of Thyl.2+, B220+ and CD4-, CDS-T cells, which both express the Fas antigen. Splenic B cells positive for surface IgM and/or surface IgD, and resident peritoneal macrophages exhibited up-regulated expression of the Fas antigen, at much higher levels than those observed in MRL/ MpJ-+/+ (MRL/+) mice. Forms of rheumatic disease were observed in these mice, although not in C3H/HeJgld/gld mice. These forms included diffuse glomerulonephritis, granulomatous arteritis, and arthritis, and were associated with the infiltration of mononuclear cells expressing the Fas antigen. Serum levels of IgG and

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Periodate-Lysine-Paraformaldehyde Fixative a New Fixative for Immunoelectron Microscopy

Cited by 3,666 publications

References 10 publications

Electron microscopic study of HLA‐DR and monocyte/macrophage staining cells in the rheumatoid synovial membrane

Electron microscopic study of HLA‐DR and monocyte/macrophage staining cells in the rheumatoid synovial membrane

Simultaneous flow cytometric analysis of surface markers and nuclear Ki‐67 antigen in leukemia and lymphoma

Rheumatic diseases in an MRL strain of mice with a deficit in the functional Fas ligand

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