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We developed an indirect immunogold-silver staining method for detection of leukocyte cell surface antigens in cell smears. Air-dried and fixed cytocentrifuge preparations or smears of peripheral blood leukocytes were incubated with monoclonal antibodies (MAb) and colloidal gold-labeled secondary antibodies. The preparations were post-fixed and silver enhancement was performed. The smears were counterstained with May-Grunwald-Giemsa and examined in brightfield light microscopy. The morphology of the cells was well preserved. Leukocytes reacting with the MAb showed black granules on their surface membranes. The intense immunostaining and the low background allowed a rapid enumeration of the positive cells. The labeling could be detected with high sensitivity by epipolarization microscopy. This immunogold-silver staining method was used to quantify T- and B-lymphocytes and natural killer cells in buffy coat smears of normal adult blood. These lymphocyte subsets correlated well with those obtained in smears with the alkaline phosphatase-anti-alkaline phosphatase (APAAP) method and with those found by labeling of mononuclear cells in suspension with immunogold-silver staining. This immunogold-silver staining method forms a good alternative to immunoenzyme methods for study of hematologic cells. In addition, it could be a general procedure for detection of cell surface antigens in all kinds of cell smears.
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W Renmans, and
E Segers, and
V De Valck, and
K Jochmans, and
B Van Camp
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M De Waele, and
W Renmans, and
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V De Valck, and
K Jochmans, and
B Van Camp
August 1990,
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
M De Waele, and
W Renmans, and
E Segers, and
V De Valck, and
K Jochmans, and
B Van Camp
January 1992,
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M De Waele, and
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January 1988,
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M De Waele, and
W Renmans, and
E Segers, and
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K Jochmans, and
B Van Camp
January 1988,
Annals of the New York Academy of Sciences,
M De Waele, and
W Renmans, and
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V De Valck, and
K Jochmans, and
B Van Camp
May 1988,
The British journal of dermatology,
M De Waele, and
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V De Valck, and
K Jochmans, and
B Van Camp
July 1996,
Histochemistry and cell biology,
M De Waele, and
W Renmans, and
E Segers, and
V De Valck, and
K Jochmans, and
B Van Camp
January 1984,
Histochemistry,
M De Waele, and
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V De Valck, and
K Jochmans, and
B Van Camp
June 1997,
Diagnostic cytopathology,
M De Waele, and
W Renmans, and
E Segers, and
V De Valck, and
K Jochmans, and
B Van Camp
March 1983,
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
