用于DNA和核抗原（Ki-67或溴脱氧尿苷）流式细胞术分析的未固定细胞核免洗双重染色法。

Washless double staining of unfixed nuclei for flow cytometric analysis of DNA and a nuclear antigen (Ki-67 or bromodeoxyuridine).

作者信息

Larsen J K, Christensen I J, Christiansen J, Mortensen B T

机构信息

Finsen Laboratory, Rigshospitalet, University Hospital, Copenhagen, Denmark.

出版信息

Cytometry. 1991;12(5):429-37. doi: 10.1002/cyto.990120508.

DOI:10.1002/cyto.990120508

PMID:1718672

Abstract

Washless methods for double staining of nuclear antigen and DNA in unfixed nuclei were compared with established methods for staining of fixed cells. The methods were tested on phytohemagglutinin (PHA)-stimulated normal human blood lymphocytes for the double staining of 1) Ki-67 antigen and DNA and 2) bromodeoxyuridine (BrdUrd) and DNA, in continuously BrdUrd-labeled cells. With respect to the discrimination between antigen-positive and -negative subpopulations, there was no statistically significant differences between the results from direct (Ki-67) or indirect (Ki-67 or BrdUrd) washless staining of unfixed nuclei and the results from staining of fixed cells. Washless staining of unfixed nuclei was found to be rapid and simple and resulted in greater precision of the DNA analysis and in less aggregation and loss of cells.

摘要

将未固定细胞核中核抗原和DNA双重染色的免洗方法与固定细胞染色的既定方法进行了比较。这些方法在植物血凝素（PHA）刺激的正常人血淋巴细胞上进行了测试，用于1）Ki-67抗原和DNA以及2）溴脱氧尿苷（BrdUrd）和DNA的双重染色，在持续BrdUrd标记的细胞中。关于抗原阳性和阴性亚群之间的区分，未固定细胞核的直接（Ki-67）或间接（Ki-67或BrdUrd）免洗染色结果与固定细胞染色结果之间没有统计学上的显著差异。发现未固定细胞核的免洗染色快速简单，并且在DNA分析中具有更高的精度，细胞聚集和损失更少。