Method for the enumeration of micronucleated erythrocyte populat

Details Method for the enumeration of micronucleated erythrocyte populat Method for the enumeration of micronucleated erythrocyte populat

1996-09-06

1999-01-12

Sisson, Bradley L.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 71, 435 72, 435 795, 436501, 436513, 436 94, 536 231, 935 76, 935 77, C12Q 168, G01N 3353, C07H 2102, C12N 1500

Patent

active

058586679

ABSTRACT:
A single laser flow cytometric method for the enumeration of micronuclei in erythrocyte populations, wherein a sample of peripheral blood or bone marrow is obtained and the cell populations in the sample are fixed. Reticulocytes in the fixed samples are treated simultaneously with RNAse and with a fluorescent labeled antibody having binding specificity for a surface marker for erythroblasts/reticulocytes. The erythrocyte populations are then stained with a nucleic acid stain which stains DNA representing micronuclei, if present. The stained and/or labelled erythrocyte populations are then exposed to a laser beam of appropriate excitation wavelength for both the nucleic acid staining dye and the fluorescent label to produce fluorescent emission. The fluorescent emission and light scatter produced by the erythrocyte populations are detected by the flow cytometer from which is calculated the number of specific erythrocyte populations in said sample.

REFERENCES:
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patent: 5409825 (1995-04-01), Hoffman et al.
Tometsko et al., "Analysis of micronucleated cells by flow cytometry. 4. Kinetic analysis of cytogenetic damage in blood," Mutation Research, vol. 334, No. 1, pp. 9-18, Jan. 1995.
Dertinger et al., "Simple and reliable enumeration of micronucleated reticulocytes with a single-laser flow cytometer," Mutation Research, vol. 371, Nos. 3,4, pp. 283-292, Dec. 20, 1996.
Begg et al., "Cell Kinetic Analysis of Mixed Populations Using Three-Color Fluorescence Flow Cytometry," Cytometry, vol. 12, No. 5, pp. 445-454, 1991 .

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