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We have developed immunocytochemical staining methods for the simultaneous phenotypic and karyotypic characterization of individual cells. Following a mild hypotonic pretreatment, isolated cells are cytocentrifuged on poly-L-lysine coated slides, fixed in formol buffered acetone, and subsequently labeled with monoclonal antibodies utilizing indirect immunoenzymatic staining procedures with horseradish peroxidase (HRP) or alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) as second antibodies. Preparations are refixed consecutively in methanol and 45% acetic acid and counterstained with either "Stains-all" (HRP labeled preparations) or Giemsa (APAAP labeled preparations). C-banding or weak G-banding, which allows the identification of individual chromosomes, can be induced in labeled as well as unlabeled mitotic cells by Ba(OH)2 and/or 2 X SSC treatment after refixation, respectively. Our method has been successfully tested with a variety of monoclonal antibodies against lymphoid, myeloid, erythropoietic, and thrombopoietic cell surface antigens. It is fast, allows the adjustment of the intensity of cell surface staining, and results in permanent preparations suitable for light microscopic analysis.
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
January 1973,
Methods in cell biology,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
March 1990,
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
June 1996,
The Histochemical journal,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
January 1990,
Haematology and blood transfusion,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
January 1986,
Histochemistry,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
January 1969,
Archiv fur experimentelle Veterinarmedizin,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
July 1985,
Diagnostic microbiology and infectious disease,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
January 1976,
Journal of immunological methods,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
June 1984,
Journal of clinical pathology,
O A Haas, and
U Köller, and
P Ambros, and
R Kornmüller, and
O Majdic, and
H Gadner, and
W Knapp
December 1987,
Nihon Ketsueki Gakkai zasshi : journal of Japan Haematological Society,
