Discriminating cellular heterogeneity using microwell-based RNA cytometry

被引：34

作者：

Dimov, Ivan K. ^{[1
,2
]}

Lu, Rong ^{[2
]}

Lee, Eric P. ^{[1
,2
]}

Seita, Jun ^{[2
]}

Sahoo, Debashis ^{[2
]}

Park, Seung-min ^{[1
,3
]}

Weissman, Irving L. ^{[2
]}

Lee, Luke P. ^{[1
]}

机构：

[1] Univ Calif Berkeley, Dept Bioengn, Berkeley Sensor & Actuator Ctr, Biomol Nanotechnol Ctr, Berkeley, CA 94720 USA

[2] Stanford Univ, Sch Med, Inst Stem Cell Biol & Regenerat Med, Stanford, CA 94305 USA

[3] Stanford Univ, Sch Med, Dept Radiol, Stanford, CA 94305 USA

来源：

NATURE COMMUNICATIONS | 2014年 / 5卷

关键词：

HEMATOPOIETIC STEM-CELLS; POLYMERASE-CHAIN-REACTION; GENE-EXPRESSION; MESSENGER-RNA; SELF-RENEWAL; REVEALS; COMPARTMENT; MICROARRAY; MECHANISM; CD41;

D O I：

10.1038/ncomms4451

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：

Discriminating cellular heterogeneity is important for understanding cellular physiology. However, it is limited by the technical difficulties of single-cell measurements. Here we develop a two-stage system to determine cellular heterogeneity. In the first stage, we perform multiplex single-cell RNA cytometry in a microwell array containing over 60,000 reaction chambers. In the second stage, we use the RNA cytometry data to determine cellular heterogeneity by providing a heterogeneity likelihood score (HLS). Moreover, we use Monte-Carlo simulation and RNA cytometry data to calculate the minimum number of cells required for detecting heterogeneity. We apply this system to characterize the RNA distributions of ageing-related genes in a highly purified mouse haematopoietic stem cell population. We identify genes that reveal novel heterogeneity of these cells. We also show that changes in expression of genes such as Birc6 during ageing can be attributed to the shift of relative portions of cells in the high-expressing subgroup versus low-expressing subgroup.

引用

页数：12

共 37 条

[1]

[Anonymous], 2006, Pattern recognition and machine learning

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