Allison D C, Mayall B H, Levin J
Department of Surgery, VA Medical Center, Baltimore, Maryland.
Cytometry. 1988 Nov;9(6):573-8. doi: 10.1002/cyto.990090610.
After staining with the Feulgen reaction, the DNA stain contents of 155 mouse bone marrow cells and 22 adjacent chicken erythrocytes were measured by absorption image cytometry by utilizing two different systems--a scanning cytometer and a video cytometer. In the scanning cytometer (M85 microdensitometer, Vickers Instruments, Malden, MA), a spot of light was scanned across the cell. In the video cytometer (TAS Plus, E. Leitz, Rockleigh, NJ), the microscope field, which may contain several nuclei, was imaged onto a Plumbicon video camera. With each system, cells were scanned, digitized into their elementary pixels, and analyzed to determine their integrated absorbance. Comparison of the DNA stain contents of the same G0/G1 bone marrow cells and chicken erythrocytes, as measured by video and scanning cytometry, showed that both techniques gave comparable results; scanning cytometry is more precise. The coefficients of variation of the measurements for the G0/G1 bone marrow cells and for the chicken erythrocytes were 5.9% and 7.0%, respectively, when measured by video cytometry at the absorption peak (584 nm), compared to 4.1% and 3.5%, respectively, for the same cells when measured by scanning cytometry off the absorption peak (615 nm). The video-based measurements were relatively lower than the scanning measurements for darkly stained cells; this suggests that glare and other optical errors which increase with stain darkness caused greater systematic errors in the video cytometer than they did in the scanning cytometer.
用福尔根反应染色后，利用两种不同系统——扫描细胞仪和视频细胞仪，通过吸收图像细胞术测量了155个小鼠骨髓细胞和22个相邻鸡红细胞的DNA染色含量。在扫描细胞仪（M85显微密度计，维氏仪器公司，马萨诸塞州马尔登）中，一束光扫描穿过细胞。在视频细胞仪（TAS Plus，徕卡公司，新泽西州罗克利）中，可能包含多个细胞核的显微镜视野被成像到氧化铅光导摄像管摄像机上。使用每个系统时，细胞被扫描、数字化成基本像素，并进行分析以确定其积分吸光度。通过视频细胞术和扫描细胞术测量的相同G0/G1期骨髓细胞和鸡红细胞的DNA染色含量比较表明，两种技术给出的结果相当；扫描细胞术更精确。在吸收峰（584nm）处通过视频细胞术测量时，G0/G1期骨髓细胞和鸡红细胞测量值的变异系数分别为5.9%和7.0%，而在吸收峰外（615nm）通过扫描细胞术测量相同细胞时，变异系数分别为4.1%和3.5%。对于深色染色的细胞，基于视频的测量相对低于扫描测量；这表明随着染色深度增加而增加的眩光和其他光学误差在视频细胞仪中比在扫描细胞仪中导致更大的系统误差。
