一种自制流式细胞仪专用溶血素对红细胞和白细胞作用的研究

目的：探讨自制流式细胞仪专用溶血素对血液红细胞的溶解效果和对淋巴细胞、单核细胞和粒细胞表面标志的影响。方法：应用流式细胞术对40份K3 EDTA抗凝外周血分别用自制流式细胞仪专用溶血素与FACS溶血素处理后，检测其对血液红细胞的溶解效果及对淋巴细胞、单核细胞和粒细胞表面特异性抗原CD14+、CD45+表达率的影响。结果：自制流式细胞仪专用溶血素和FACS溶血素，均能使红细胞彻底溶解，且不破坏白细胞，在流式细胞仪获取的前向散射光(FSC)和侧向散射光(SSC)散点图中，可见清晰的淋巴细胞、单核细胞和粒细胞群，其CD14+表达率分别为(1.01±0.24)%和(1.00±0.23)%、(92.80±1.98)%和(92.81±1.94)%、(91.16±3.68)%和(91.22±3.76)%；CD45+表达率分别为(98.94±0.71)%和(98.90±0.72)%、(95.15±2.73)%和(95.11±2.76)%、(99.39±0.48)%和(99.41±0.47)%，使用两种溶血素检测得到的白细胞表面标志具有很好的一致性，其差异无统计学意义（P＞0.05）。结论：自制流式细胞仪专用溶血素具有很好的红细胞溶解效果，但不破坏白细胞，且对淋巴细胞、单核细胞和粒细胞群表面抗原的表达无明显影响作用。

A self-made lysing solution in flow cytometry on white blood cells and red blood cells

To investigate the effect of a self made lysing solution in flow cytometry on red blood cells, and on surface markers of lymphocytes, monocytes and granulocytes. Methods： K3 EDTA peripheral blood of 40 healthy volunteers was dealt with a self made lysing solution and a FACS Lysing solution, and then the lysing effect on red blood cells and the influence on CD14+， CD45+ on lymphocytes, monocytes and granulocytes were detected. Results： Both the self made lysing solution and the FACS Lysing solution could lyse red blood cells completely, but not destroy leukocytes. In FSC/SSC dot plot, there were three clusters of cells: lymphocytes, monocytes and granulocytes. The expression of CD14+ on these cells were (1.01±0.24)% and (1.00±0.23)%, (92.80±1.98)% and (92.81±1.94)%, (91.16±3.68)% and (91.22±3.76)% respectively； and the expression of CD45+ on these cells were (98.94±0.71)% and (98.90±0.72)%, (95.15±2.73)% and (95.11±2.76)%, (99.39±0.48)%and (99.41±0.47)% respectively. The expression of surface markers on leukocytes was not significantly different between the self made lysing solution and the FACS lysing solution(P＞0.05). Conclusion： The self made lysing solution can lyse red blood cells completely ,but not destroy leukocytes and influence the expression of surface antigens on lymphocytes, monocytes and granulocytes significantly.