Accurate assessment of cell count and viability with a flow cytometer.
BACKGROUND
In this study we developed a method to measure cell concentration and viability in specimens received in flow cytometry and cytogenetics laboratories.


METHODS
Specimens are stained with a vital fluorescent dye, SYTO13, the cell impermeant viability dye, 7-AAD, and a leukocyte marker, CD45. After the addition of an internal calibrator microsphere, FLOW-COUNT, the flow cytometer is capable of measuring the viability of nucleated cells, giving a general assessment of leukocyte populations and measuring their concentration.


RESULTS
An accurate assessment of specimen quality is an important parameter when performing flow cytometric and cytogenetic leukemia/lymphoma assessment. High quality specimen is desired to avoid the pitfalls of non-specific staining and limited cellularity/viability.


CONCLUSIONS
Use of a cell count and viability measurement prior to leukemia and lymphoma assessment by flow cytometry and cytogenetics helps to increase the rate of successful immunophenotypic and cytogenetic analysis.
Shenkin
Mark
M
AmeriPath Inc., Orlando, FL, USA. mshenkin@ameripath.com
Babu
Ramesh
R
Maiese
Russell
R
D015415:Biomarkers / Q000032:analysis; D002452:Cell Count / Q000295:instrumentation / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D003582:Cytogenetics / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D007202:Indicators and Reagents; D007938:Leukemia / Q000175:diagnosis* / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000032:analysis; D008223:Lymphoma / Q000175:diagnosis* / Q000276:immunology
D015415:Biomarkers; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D017493:Leukocyte Common Antigens