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探讨了改良前后两种MTT比色法的显色机制及区别,并进行了改良方法与未改良方法及同位素掺入方法检测敏感性的比较和改良方法实验影响因素的观察。结果表明①改良前异丙醇只能溶解细胞外部甲,改良后异丙醇不但能溶解细胞周围的甲,而且能将细胞内的MTT代谢物全部溶解。②改良后较改良前的敏感性有了较大的提高,其敏感性检测结果与~3H-TdR检测结果相一致。③改良方法可以测出约3×10~3个CTLL_2细胞和用6×10~3个细胞活化后的PBL。④适当提高MTT浓度能进一步提高检测敏感性及OD值。
The coloration mechanisms and differences between the two MTT colorimetry methods before and after the improvement were discussed. The sensitivity of the improved method and the unmodified method and the isotope incorporation method were compared, and the influencing factors of the improved method experiment were observed. The results showed that 1 before the improvement of isopropanol can only dissolve the outer layer of the cell. After the improvement, isopropanol can not only dissolve the formazan around the cell, but also completely dissolve the MTT metabolites in the cell. 2 The sensitivity of the improved method before the improvement has been greatly improved. The sensitivity test results are consistent with the results of ~3H-TdR detection. 3 Improved methods can detect about 3×10~3 CTLL 2 cells and PBL activated with 6×10~3 cells. 4 Proper increase of MTT concentration can further increase detection sensitivity and OD value.