Durand R E
J Histochem Cytochem. 1982 Feb;30(2):117-22. doi: 10.1177/30.2.6174559.
Hoechst 33342 staining of multicell spheroids, three-dimensional cell clusters grown in vitro, results in a marked gradient of cellular fluorescent intensities inward from the spheroid periphery. The penetration of the dye is concentration and time dependent, so staining can be coupled with fluorescence activated cell sorting techniques to allow disaggregated single cells to be sorted or selected according to their degree of staining and therefore their depth within the spheroid. We have found the staining procedure to be highly reproducible, and to result in minimal toxicity even to the more brightly staining external cells. Comparison of this technique with others for cell selection suggests that increased resolution is available with the Hoechst technique.
用Hoechst 33342对多细胞球体（体外培养的三维细胞簇）进行染色，会导致从球体周边向内部出现明显的细胞荧光强度梯度。染料的渗透取决于浓度和时间，因此染色可与荧光激活细胞分选技术相结合，以便根据细胞的染色程度以及它们在球体内的深度对解离后的单细胞进行分选或筛选。我们发现该染色程序具有高度的可重复性，并且即使对染色较亮的外部细胞也只会产生最小的毒性。将该技术与其他细胞分选技术进行比较表明，Hoechst技术具有更高的分辨率。
