Open AccessTechnique and staining optimization leucoconcentration
Author(s) -
Pierrez J.,
Guerci A.,
Guerci O.
Publication year - 1987
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990080515
Subject(s) - staining , flow cytometry , lysis , picric acid , microbiology and biotechnology , lysis buffer , cell , chemistry , cytometry , chromatography , biology , pathology , biochemistry , medicine , organic chemistry
In cytometric clinical application, it is important to obtain cell suspensions rapidly with as little cytological alteration as possible. A procedure has been achieved to prepare cell suspensions for flow cytometric analysis. The leucoconcentration technique, first described by Herbeuval for cytologic analysis, has been modified to be applied in cytometry. This technique involves Saponin lysis of red cells of peripheral blood or bone marrow samples that have been previously fixed with picric acid alcohol solution. Cells in suspension are not shifted and tinctorial affinity is not modified. Then cells have been stained with Mithramycin. Each parameter defined by Crissman has been analyzed to define the best staining conditions. The availability of Leucoconcentration with Mithramycin‐DNA‐staining permits determination of cell cycle with a fine resolution.