Oxazine stained lymphocytes and method

Details Oxazine stained lymphocytes and method Oxazine stained lymphocytes and method

1994-02-09

1995-04-18

Snay, Jeffrey R.

Chemistry: analytical and immunological testing

Biological cellular material tested

424 3, 424 71, 424722, 435 2, 4352402, G01N 3348

Patent

active

054077940

ABSTRACT:
Peripheral blood lymphocytes displayed a plurality of sizes and colors when exposed first to a fixative solution of C.I. basic blue 141, then to an aqueous alkaline solution of the same dye and subsequently treated with a neutral HEPES buffer solution containing small but effective amounts of various metal compounds. As confirmed with purified lymphocyte subpopulations obtained with a cell sorter, T helper cells (CD4) were small and their nuclei and cytoplasm stained deep blue. T cytotoxic/suppressor cells (CDS) were larger than T helper cells, their nuclei stained pale green or blue green and their cytoplasm contained a cluster of magenta colored granules. Used in the process described, basic blue 141 provides means of identifying and differentiating CD4 and CD8 cells under the ordinary light microscope without using monoclonal antibodies or fluorescence.

REFERENCES:
patent: 4376820 (1983-03-01), Gianinni et al.
patent: 4400370 (1983-08-01), Kass
patent: 4500509 (1985-02-01), Kass
Sigma 1990, "Biochemicals Organic Compounds . . . " p. 1496.
American Journal of Clinical Pathology 88:436-441, 1987. "Identification of Neutrophils . . . "Kass.
Rapid Identification of T Helper and T Cytotoxic/Suppressor Lymphocytes with Oxazine Dye Lawrence Kass Departments of Pathology and Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio 44109 1052-0295/93/6805-247/$3.00/0 Biotechnic & Histochemistry Copyright .COPYRGT.1993 by Williams & Wilkins vol. 68 No. 5 published Oct. 4, 1993 pp. 247-254.

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