DNA flow cytometry of paraffin-embedded tissue.
DNA flow cytometry of paraffin-embedded tissue.
复制标题
石蜡包埋组织的 DNA 流式细胞仪。
DOI：
10.1002/cyto.990050618
复制
发表时间：
1984
期刊：
影响因子：
--
通讯作者：
E. Musgrove
中科院分区：
文献类型：
--
作者：
D. Hedley;M. Friedlander;I. Taylor;C. Rugg;E. Musgrove
去年，我们发表了一种使用流式细胞术分析石蜡包埋组织细胞DNA含量的方法（1）。许多人写信给我们，表达了对这种方法的兴趣，但也要求澄清某些技术要点，因为我们现在有使用这种方法的丰富经验，我们认为现在是时候讨论一下原始论文中没有完全讨论的一些问题了。我们已经测试了一系列固定剂，发现中性福尔马林或福尔马林醋酸产生最好的效果。布因的固定剂不太令人满意，可能是由于苦味酸的荧光淬灭，而汞基固定剂是流式细胞术中最差的。使用中性福尔马林的固定质量可能也很关键，因为在死后检查中获得的包埋材料产生非常差的结果，我们现在认为原始材料的处理方式是无法获得可解释的DNA直方图的主要原因。
Last year we published a method for analyzing cellular DNA content of paraffin-embedded tissue using flow cytometry (1). A number of people have written to us expressing interest in the method but also requesting clarification of certain technical points, and since we now have extensive experience using this approach, we thought it timely to go over some points not fully dealt with in the original paper.The way in which the tissue was fixed is of importance. We have tested a range of fixatives and have found that neutral formalin or formaldehydelacetonel acetic acid produces the best results. Bouin’s fixative is less satisfactory, probably owing to fluorescence quenching by the picric acid, and mercury-based fixatives are the worst of all for flow cytometry. Using neutral formalin the quality of fixation is probably also critical, since embedded material taken at postmortem examination produces very poor results, and we now think that the way in which the original material was handled is the major cause for inability to obtain an interpretable DNA histogram.