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Improving single-cell transcriptome sequencing efficiency with a microfluidic phase-switch device
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DOI：10.1039/c9an00823c.png)
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摘要
En 中文
In this paper, we present a novel method to improve the efficiency of single-cell transcriptome sequencing for analyzing valuable cell samples. The microfluidic device we designed integrates multiple singlecell isolation chambers with hydrodynamic traps and achieves a nearly 100% single-cell capture rate and minimal cell loss, making it particularly suitable for samples with limited numbers of cells. Single cells were encapsulated using a novel phase-switch method into picoliter-sized hydrogel droplets and easily recovered for subsequent reactions. Minimizing the reaction volume resulted in a high reverse transcription (RT) efficiency for RNA sequencing (RNA-Seq). With this novel microfluidic platform, we captured dozens of hESCs (H9) simultaneously and obtained live cells in individual picoliter volumes, thus allowing for the convenient construction of a high-quality library for deep single-cell RNA-Seq. Our single-cell RNA-Seq results confirmed that a spectrum of pluripotency existed within an H9 colony. This integrated microfluidic platform can be applied to various cell types for the investigation of rare cellular events, and the phase-switch single-cell processing strategy will improve the efficiency and accessibility of single-cell transcriptome sequencing analysis.
Keyword:
CIRCULATING TUMOR-CELLS
EMBRYONIC STEM-CELLS
GENE-EXPRESSION
HETEROGENEITY
LINES
PLURIPOTENCY
MAINTAIN
GROWTH
DNA
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期刊
A
IF:
3.3
论文数:
1.6W
被引数:
3.3W
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