SENSITIVE DETECTION OF RNAS IN SINGLE CELLS BY FLOW-CYTOMETRY

被引：31

作者：

YU, H ^{[1
]}

ERNST, L ^{[1
]}

WAGNER, M ^{[1
]}

WAGGONER, A ^{[1
]}

机构：

[1] CARNEGIE MELLON UNIV,CTR LIGHT MICROSCOPE IMAGING & BIOTECHNOL,4400 5TH AVE,PITTSBURGH,PA 15213

来源：

NUCLEIC ACIDS RESEARCH | 1992年 / 20卷 / 01期

关键词：

D O I：

10.1093/nar/20.1.83

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A rapid and sensitive fluorescent in situ hybridization method has been developed to probe RNA contents of individual cells by flow cytometry. Fixed cells in suspension were hybridized with 5' end-fluorophore-labeled oligodeoxynucleotides complementary to defined regions of the RNA of interest and analyzed by flow cytometry. With this method, we monitored combinations of histone H4 mRNA, 18S rRNA and 28S rRNA levels in synchronized HeLa S3 cells by multicolor analysis. A fluorescence signal equivalent to 1800 copies of histone H4 mRNA per cell was detected with signal-to-background ratio of 5.4. If non-specific binding of the fluorophore-labeled probe can be reduced, as few as 100 copies of mRNA of the size of H4 could be detected in individual cells by flow cytometry.

引用

页码：83 / 88

页数：6

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