U. Oelschlaegel et al., Flow cytometric DNA-quantification of three-color immunophenotyped cells for subpopulation specific determination of aneuploidy and proliferation, J IMMUNOL M, 253(1-2), 2001, pp. 145-152
A method is described for three-color immunophenotyping and simultaneous DN
A-quantification using a flow cytometer equipped with a 488-nm argon laser
and a mercury lamp (UV). The approach includes reproducible immunophenotypi
ng comparing antigen expression before and after cell manipulation for DNA-
measurement. The coefficients of variation after DNA-staining (CV = 3.13 fo
r T-cells in peripheral blood and CV = 3.38 for T-cells in bone marrow) wer
e adequate for exact DNA-analysis. For aneuploidy detection, a true interna
l standard was established measuring, for example, the DNA-content of T-cel
ls in B-cell disease simultaneously with the DNA-content of the malignant c
ells. Using this method, aneuploidies could be unequivocally detected in 17
out of 24 patients with multiple myeloma. Furthermore, intratumor heteroge
neities in DNA-content and antigen expression could be recognized, allowing
an exact separation of tumor cells and normal hematopoiesis. The study als
o demonstrated the importance of exact immunophenotypic characterization of
lymphocyte subpopulations and the determination of their specific prolifer
ation, for example after proliferation induction in cell cultures. Future s
tudies should address the applicability of this rather simple multiparamete
r approach for simultaneous immunophenotyping and DNA-measurement especiall
y in the detection of minimal amounts of aneuploid cells after chemotherapy
. (C) 2001 Elsevier Science B.V. All rights reserved.