Quantification of virus-infected cells using RNA FISH-Flow

被引:4
|
作者
Warren, Cody J. [1 ]
Barbachano-Guerrero, Arturo [2 ]
Huey, Devra [1 ]
Yang, Qing [2 ]
Worden-Sapper, Emma R. [2 ]
Kuhn, Jens H. [3 ]
Sawyer, Sara L. [2 ]
机构
[1] Ohio State Univ, Dept Vet Biosci, Columbus, OH 43210 USA
[2] Univ Colorado, BioFrontiers Inst, Dept Mol Cellular & Dev Biol, Boulder, CO 80303 USA
[3] Natl Inst Allergy & Infect Dis, NIH, Integrated Res Facil Ft Detrick, Frederick, MD 21702 USA
来源
STAR PROTOCOLS | 2023年 / 4卷 / 02期
关键词
In Situ Hybridization; Microbiology; Molecular Biology;
D O I
10.1016/j.xpro.2023.102291
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We present a protocol to detect cells that have been infected by RNA viruses. The method, RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow), uses 48 fluorescently labeled DNA probes that hybridize in tandem to viral RNA. RNA FISH-Flow probes can be synthesized to match any RNA virus genome, in either sense or anti-sense, enabling detection of genomes or replication intermediates within cells. Flow cytometry enables high-throughput analysis of infection dynamics within a population at the single cell level. For complete details on the use and execution of this protocol, please refer to Warren et al. (2022).(1)
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页数:18
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