PICOGRAM PER CELL DETERMINATION OF DNA BY FLOW CYTOFLUOROMETRY
PICOGRAM PER CELL DETERMINATION OF DNA BY FLOW CYTOFLUOROMETRY
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DOI：
10.1016/0003-2697(84)90374-9
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
RASCH, EM
中科院分区：
文献类型：
--
作者：
LEE, GM;THORNTHWAITE, JT;RASCH, EM
使用从< 0.2 g组织或107个细胞中分离的细胞核，提出了一种在pg水平上定量测定每个细胞DNA量的方法。该技术是基于4“，6-二氨基-2-苯基吲哚（DAPI）与DNA结合时的荧光增强。一个快速，一步的核分离和DNA染色程序是用来准备组织样品流式细胞术分析。冷冻组织提供的结果与新鲜组织相当。用鸡和鳟鱼的红细胞核作为参考标准，测定了几种哺乳动物（大鼠、兔、小鼠、猫和人）和亚马逊茉莉鱼的二倍体细胞的DNA含量。从同一生物体的不同组织中获得的一致值表明了该方法用于DNA测量的准确性。
Using nuclei isolated from < 0.2 g tissue or 107 cells, a method is presented for the quantitative determination of amounts of DNA per cell at the pg level. This technique is based on the enhanced fluorescence of 4'',6-diamidino-2-phenylindole (DAPI) when it binds to DNA. A rapid, 1-step nuclear isolation and DNA staining procedure is used to prepare tissue samples for flow cytometric analysis. Frozen tissues give results comparable to those for fresh tissue. Both chicken and trout erythrocyte nuclei were used as reference standards in the determination of amounts of DNA per diploid cell for several mammals [rat, rabbit, mouse, cat and human] and Amazon molly fish. The consistent values obtained for different tissues from the same organism show the accuracy of this method for DNA measurement.