Expression profiling. Combinatorial labeling of single cells for gene expression cytometry

• Published in: Science (American Association for the Advancement of Science) Vol. 347; no. 6222; p. 1258367
• Main Authors: Fan, H Christina, Fu, Glenn K, Fodor, Stephen P A
• Format: Journal Article
• Language: English
• Published: United States 06.02.2015
• Subjects: Combinatorial Chemistry Techniques; DNA Barcoding, Taxonomic - methods; DNA, Complementary - chemistry; Flow Cytometry - methods; Gene Expression Profiling - methods; Hematopoiesis - genetics; Humans; Microspheres; RNA, Messenger - analysis; RNA, Messenger - chemistry; Single-Cell Analysis - methods; T-Lymphocytes
• ISSN: 1095-9203
• DOI: 10.1126/science.1258367

We present a technically simple approach for gene expression cytometry combining next-generation sequencing with stochastic barcoding of single cells. A combinatorial library of beads bearing cell- and molecular-barcoding capture probes is used to uniquely label transcripts and reconstruct the digital gene expression profile of thousands of individual cells in a single experiment without the need for robotics or automation. We applied the technology to dissect the human hematopoietic system and to characterize heterogeneous response to in vitro stimulation. High sensitivity is demonstrated by detection of low-abundance transcripts and rare cells. Under current implementation, the technique can analyze a few thousand cells simultaneously and can readily scale to 10,000s or 100,000s of cells.