Combinatorial labeling of single cells for gene expression cytometry

被引:330
作者
Fan, H. Christina [1 ]
Fu, Glenn K. [1 ]
Fodor, Stephen P. A. [1 ]
机构
[1] Cellular Res Inc, Palo Alto, CA 94304 USA
关键词
POLYMERASE-CHAIN-REACTION; RNA-SEQ; CELLULAR HETEROGENEITY; FLOW-CYTOMETRY; MOLECULE; IMMUNE; IDENTIFICATION; SIGNATURES; PHENOTYPE; FREQUENCY;
D O I
10.1126/science.1258367
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
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页数:9
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