CELL-CYCLE ANALYSIS BY COMBINING THE 5-BROMODEOXYURIDINE-33258 HOECHST TECHNIQUE WITH DNA-SPECIFIC ETHIDIUM-BROMIDE STAINING
CELL-CYCLE ANALYSIS BY COMBINING THE 5-BROMODEOXYURIDINE-33258 HOECHST TECHNIQUE WITH DNA-SPECIFIC ETHIDIUM-BROMIDE STAINING
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DOI：
10.1002/cyto.990020107
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
ELLWART, J
中科院分区：
文献类型：
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作者：
BOHMER, RM;ELLWART, J
细胞在含5-溴脱氧尿苷（BrdUrd）的培养基中生长，收获并用33258 Hoechst和溴化乙锭组合染色，用于双参数流式细胞仪中的分析。两种荧光都由UV激光激发，溴化乙锭还由Hoechst染料的能量转移激发。两种荧光的分离被证明是极好的，投影直方图显示出与通过单一染料染色获得的质量相当的质量。溴化乙锭荧光给出了细胞在DNA复制周期中的位置信息，而BrdUrd猝灭的Hoechst荧光给出了细胞在BrdUrd掺入开始时位于周期中的位置信息。以这种方式，获得关于细胞在BrdUrd孵育时间期间通过循环行进的距离的信息。这种方法可能成为一个强大的工具，在许多调查处理细胞周期扰动文化。
Cells were grown in 5‐bromodeoxyuridine(BrdUrd)‐containing medium, harvested and stained with a combination of 33258 Hoechst and ethidium bromide for analysis in a two‐parametrical flow cytometer. Both fluorescences were excited by UV laser light, and ethidium bromide was additionally excited by energy transfer from the Hoechst dye. The separation of the two fluorescences proved to be excellent, the projected histograms showing a quality comparable to the quality obtained by single dye staining. The ethidium bromide fluorescence gives information as to where the cell is located within the cycle of DNA replication, while the BrdUrd‐quenched Hoechst fluorescence gives information as to where in the cycle the cell was located at the beginning of BrdUrd incorporation. In this way information is obtained concerning the distance a cell traveled through the cycle during the BrdUrd incubation time. This method may become a powerful tool in many investigations dealing with cell cycle perturbations in culture.