FLOW CYTOMETRIC CELL CYCLE ANALYSIS USING THE QUENCHING OF 33258 HOECHST FLUORESCENCE BY BROMODEOXYURIDINE INCORPORATION
FLOW CYTOMETRIC CELL CYCLE ANALYSIS USING THE QUENCHING OF 33258 HOECHST FLUORESCENCE BY BROMODEOXYURIDINE INCORPORATION
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使用溴脱氧尿苷猝灭 33258 HOECHST 荧光进行流式细胞术细胞周期分析
DOI：
10.1111/j.1365-2184.1979.tb00117.x
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发表时间：
1979
影响因子：
8.5
通讯作者：
R. Bohmer
中科院分区：
文献类型：
--
作者：
R. Bohmer
L 细胞培养物在含有 2.0 mg/l 溴脱氧尿苷 (BUdR) 的培养基中生长，并用荧光染料 33258 Hoechst 染色以进行流式细胞术分析。在接触 BUdR 期间，细胞在新合成的 DNA 中用 BUdR 取代胸苷。新 DNA 无法用 33258 Hoechst 染色，而 33258 Hoechst 对胸苷具有高度特异性。在流式细胞仪中研究了向生长培养基中添加 BUdR 后荧光分布的时间发展，并且使用数据来计算指数生长培养物中 G1、S 和 G2+ M 阶段的平均持续时间以及同步培养物中的循环渡越时间。在每个实验中确定非循环细胞的百分比。
Cultures of L cells were grown in medium containing 2.0 mg/l bromodeoxyuridine (BUdR) and stained with the fluorescent dye 33258 Hoechst for flow cytometric analysis. During exposure to BUdR, the cells replace thymidine by BUdR in the newly synthesized DNA. The new DNA is not stainable with 33258 Hoechst, which is highly specific for thymidine. The temporal development of the fluorescence distributions after addition of BUdR to the growth medium has been investigated in the flow cytometer, and the data were used to calculate the mean durations of the phases G1, S and G2+ M in exponentially growing cultures as well as the cycle transit times in synchronized cultures. The percentage of non‐cycling cells was determined in each experiment.