期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 391, 期 1-2, 页码 133-145出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2013.03.002
关键词
Single cell; Gene expression; T-cell activation; qPCR
资金
- NIAID, NIH
- Collaboration for AIDS Vaccine Discovery (CAVD) from the Bill & Melinda Gates Foundation [OPP1032325]
Highly multiplexed, single-cell technologies reveal important heterogeneity within cell populations. Recently, technologies to simultaneously measure expression of 96 (or more) genes from a single cell have been developed for immunologic monitoring. Here, we report a rigorous, optimized, quantitative methodology for using this technology. Specifically: we describe a unique primer/probe qualification method necessary for quantitative results; we show that primers do not compete in highly multiplexed amplifications; we define the limit of detection for this assay as a single mRNA transcript; and, we show that the technical reproducibility of the system is very high. We illustrate two disparate applications of the platform: a bulk approach that measures expression patterns from 100 cells at a time in high throughput to define gene signatures, and a single-cell approach to define the coordinate expression patterns of multiple genes and reveal unique subsets of cells. Published by Elsevier B.V.
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