Identification of L3 leukemia and Burkitt's lymphoma cells by flow cytometric quantitation of nuclear and cellular RNA and DNA content.
Separate quantitations of nucleic acids of isolated nuclei and cells of L3 leukemia/Burkitt's lymphoma cell populations of peripheral blood (PB), bone marrow (BM) and lymph node (LN) cell suspensions of 17 patients were performed by acridine orange (AO) flow cytometry (FCM). The cell populations were analysed with respect to cell cycle characteristics and DNA/RNA distribution histograms of cells in various compartments of the cell cycle using mean value, coefficient of variation (CV), third moment about the mean as a measure of skewness (MOM3). The correlations between numbers of L3 blasts detected by microscopy and aneuploid cells quantified by FCM in 10 patients with aneuploid disease were r = 0.75 and r = 0.85 for BM and PB cell populations, respectively. Content of both nuclear and cellular RNA was 3-4 times higher in L3 cells than in normal donor lymphocytes. Percentages of cells in SG2M phase of L3 populations were significantly different from control populations in BM (p less than 0.01) and PB (p less than 0.0001). All samples with L3 blasts had abnormal CV of the DNA, and abnormal CV and MOM3 of the RNA peaks of G0/1 cell histograms. All samples with no blasts by morphology had normal DNA and RNA patterns. AO FCM correctly classified all specimens as involved or not involved with disease in accordance with the cytomorphological diagnoses. Combining AO FCM with prior cell separation increased the sensitivity of the method to detect abnormal cells to 0.02%, or 0.5 L3 cells per microliter of blood.[1]References
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