Flow cytometric fluorescence emission spectrum analysis of Hoechst-33342-stained DNA in chicken thymocytes.
Flow cytometric fluorescence emission spectrum analysis of Hoechst-33342-stained DNA in chicken thymocytes.
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鸡胸腺细胞中 Hoechst-33342 染色 DNA 的流式细胞术荧光发射光谱分析。
DOI：
10.1002/cyto.990060406
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Smith,PJ
中科院分区：
文献类型：
--
作者：
Watson,JV;Nakeff,A;Chambers,SH;Smith,PJ
在我们定制的流式细胞仪中，在两个荧光波长带（绿色和紫色）上同时分析Hoechst-33342染色的鸡胸腺细胞，并鉴定了两个主要亚群。在一个子集（总数的33%）中，发射光谱随时间保持恒定，各自的绿色和紫色荧光强度几乎没有变化。在另一个子集（占总数的42%）中，染色期间绿色荧光增加，由于荧光发射的“形状”随时间变化，导致绿色-紫色比发生相当大的变化。数据表明，两个结合位点，或在同一位点的两种类型的结合，存在于DNA中的这种染料，这些具有不同的结合能，因此，不同的荧光发射特性。
Hoechst‐33342‐stained chicken thymocytes were analysed simultaneously on two fluorescence wavelength bands (green and violet) in our custom‐built flow cytometer, and two major subsets were identified. In one subset (33% of the total) the emission spectrum remained constant with time, with little change in the respective green and violet fluorescence intensities. In the other subset (42% of the total) the green fluorescence increased during staining, resulting in a considerable change in the green‐to‐violet ratio, due to a change in the “shape” of the fluorescence emission with time. The data indicate that two binding sites, or two types of binding at the same site, exist in DNA for this dye and that these have different binding energies and, consequently, different fluorescence emission properties.