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目的:比较磺基罗丹明B(SRB)法与二苯基溴化四氮盐(MTT)比色法细胞计数的灵敏性和稳定性。方法:取4种不同类型对数生长期的细胞,调整细胞的密度为1×106/L,并按比例稀释为0.039×105~10×105个细胞的梯度。(1)用SRB法与MTT比色法测定各细胞梯度相应A值,分别分析两种方法测到的A值与相应细胞梯度细胞数的相关性。(2)SRB法与MTT比色法各随机取100个A值,测量其在2、16、24h及7d后的A值,比较这两种方法的稳定性。结果:SRB法和MTT比色法测定的A值与不同细胞系的细胞数目均有极好的相关性。SRB法测定的结果几乎不随时间而变化;而MTT法测定的结果受时间影响较大。结论:SRB法与MTT比色法相比较更适于进行大规模的细胞计数。
OBJECTIVE: To compare the sensitivity and stability of cell counting by sulfobarodamine B (SRB) and diphenyltetrazolium (MTT) colorimetric methods. Methods: Four different types of cells in logarithmic growth phase were used. The cell density was adjusted to 1 × 106 / L and the cells were diluted to a gradient of 0.039 × 105-10 × 105 cells. (1) The corresponding A value of each cell gradient was determined by SRB method and MTT colorimetric method. The correlation between the A value measured by the two methods and the cell number of the corresponding cell gradient was analyzed respectively. (2) SRB method and MTT colorimetry were randomly selected 100 A value, measured at 2,16,24h and 7d after the A value, compare the stability of the two methods. Results: The values of A determined by SRB and MTT assay were highly correlated with the number of cells in different cell lines. The result of SRB method hardly changed with time; but the result of MTT method was greatly affected by time. Conclusion: Compared with MTT colorimetric method, SRB method is more suitable for large-scale cell counting.