Highly multiplexed quantitation of gene expression on single cells.

Publication Type:

Journal Article

Authors:

Dominguez, Maria H; Chattopadhyay, Pratip K; Ma, Steven; Lamoreaux, Laurie; McDavid, Andrew; Finak, Greg; Gottardo, Raphael; Koup, Richard A; Roederer, Mario

Source:

Journal of immunological methods, Volume 391, Issue 1-2, p.133-45 (2013)

Keywords:

2013, April 2013, Public Health Sciences Division, Vaccine and Infectious Disease Division

Abstract:

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.