[Flow cytometry in determination of DNA content of lymphoid cells: study of methodology and its preliminary clinical application].
The methodology of flow cytometry (FCM) in the determination of DNA content of lymphoid cells was studied. We conclude that the optimal conditions for the Ortho Instruments L-30 Cytofluorograf flow cytometer are as follows: 70% ethanol should be used as fixative and propidium iodide as DNA fluorescent stain at a final concentration of 50 micrograms/ml. RNase and Triton-X-100 added to the mixture and the use of fresh chicken red blood cells (CRBC) as internal standard will ensure the reliability of the data. The interference of doublet cells is eliminated by selecting single cell area signals. We discovered that aneuploidy detection can be optimized by using the coefficient of variation ratio (CVR) together with the DNA ratio (DR). A special computer program was written to calculate the percentage of cells in different phases of the cell cycle, according to DNA histograms derived from FCM. Based on assessment of the DNA content of lymphoid cells of 33 normal subjects, the normal ranges of DR, CVR and S% were found to be 2.57-2.83, 1.02 and less than 9.6% respectively. The diagnosis of two cases of lymphoid cell malignancy is reported and the potential of this method for wide clinical application discussed.
Zhang
Z G
ZG
Song
S Z
SZ
D004247:DNA / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis*; D005434:Flow Cytometry; D006801:Humans; D008214:Lymphocytes / Q000032:analysis; D008223:Lymphoma / Q000175:diagnosis; D008258:Waldenstrom Macroglobulinemia / Q000175:diagnosis
D004273:DNA, Neoplasm; D004247:DNA