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本文比较了用溶血空斑技术计算小白鼠单个抗体分泌细胞数和用光电比色法测定小鼠脾细胞抗体分泌量的方法。由8个经绵羊红细胞免疫的小鼠脾细胞混合制成的悬液,准确计数后制成每ml含1×10~7及2×10~6细胞数,然后由每个稀释度分别测定PFC及溶血素OD值。PFC测定重复20个平皿,而溶血素OD值测定重复10管。由此计算出两种方法所得数字的标准差与标准误。结果指出,用光电比色测溶血素OD值的方法的准确度要比溶血空斑测定法高2.7—2.8倍。但根据同一试验结果,也发现光电比色法的不足之处,当所用免疫脾细胞数相差5倍时(1×10~7,2×10~6),PFC值相差约6.5倍,而OD值之差不到一倍。就此点看来,光电比色法虽较准确,但其敏感度不如溶血空斑技术。因此单独使用溶血素OD值为测定B细胞功能的指标,则不够理想。溶血空斑测定法虽人为影响因素较多,但如严格控制条件,重复性仍较好。
In this paper, the number of single antibody secreting cells in mice was calculated by hemolytic plaque technique and the amount of antibody secreted by splenocytes in mice was measured by photoelectric colorimetry. A suspension of 8 mouse spleen cells immunized with sheep erythrocytes was accurately counted and then made into 1 × 10 7 cells and 2 × 10 6 cells per ml, and then each dilution was used to measure the PFC And hemolysin OD value. PFC assay was repeated 20 plates, and the determination of hemolysin OD value of 10 tubes repeated. From this we can calculate the standard deviation and standard error of the figures obtained by the two methods. The results show that the use of photoelectric colorimetric hemolysin OD value of the accuracy of hemolytic plaque assay 2.7-2.8 times higher. However, according to the same test result, the deficiency of photoelectric colorimetric method was also found. When the number of immune spleen cells used was 5 times different (1 × 10 ~ 7, 2 × 10 ~ 6) Less than double the value. In this point of view, the photoelectric colorimetric method is more accurate, but its sensitivity is not as good as hemolytic plaque technology. Therefore, the separate use of hemolysin OD value for the determination of B cell function index, less than ideal. Hemolytic plaque assay, although more influencing factors of man-made, but if strict control conditions, repeatability is still good.