Single-Cell Multiparametric Analysis of Rare HIV-Infected Cells Identified by Duplexed RNAflow-FISH.
Single-Cell Multiparametric Analysis of Rare HIV-Infected Cells Identified by Duplexed RNAflow-FISH.
复制标题
通过双链 RNAflow-FISH 鉴定罕见 HIV 感染细胞的单细胞多参数分析。
DOI：
10.1007/978-1-0716-1871-4_20
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Kaufmann,DanielE
中科院分区：
文献类型：
--
作者：
Dubé,Mathieu;Kaufmann,DanielE
HIV-infected cells are difficult to characterize in vivo because of their great paucity and their diversity. This chapter describes a duplexed flow cytometry method that enables detection, quantification and phenotyping of these rare cells at single-cell resolution. Primary CD4+T cells are enriched from PBMCs, stained for surface and intracellular proteins and then subjected to fluorescent in situ hybridization to label viral RNA before acquisition on a flow cytometer. Technical and analytical advices are provided to improve the quality of the data. This flow cytometric RNA fluorescent in situ hybridization (RNAflow-FISH) procedure can be applied to the characterization of both HIV-infected cells from viremic people living with HIV and reactivated viral reservoirs from virally suppressed individuals on therapy.
