Single-cell analysis and sorting using droplet-based microfluidics.
Single-cell analysis and sorting using droplet-based microfluidics.
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DOI：
10.1038/nprot.2013.046
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发表时间：
2013-05
期刊：
影响因子：
14.8
通讯作者：
Heyman, John A.
中科院分区：
文献类型：
--
作者：
Mazutis, Linas;Gilbert, John;Ung, W. Lloyd;Weitz, David A.;Griffiths, Andrew D.;Heyman, John A.
我们提出了基于液滴的微流体方案，用于对单细胞的高通量分析和分类单细胞的分类。 - 激活细胞分类。用抗小鼠IgG抗体覆盖在50 pl液滴中。 200 Hz和细胞富集。 H;整个过程，包括微流体设备和哺乳动物细胞的制备，需要5-7 d。
We present a droplet-based microfluidics protocol for high-throughput analysis and sorting of single cells. compartmentalization of single cells in droplets enables the analysis of proteins released from or secreted by cells, thereby overcoming one of the major limitations of traditional flow cytometry and fluorescence-activated cell sorting. as an example of this approach, we detail a binding assay for detecting antibodies secreted from single mouse hybridoma cells. secreted antibodies are detected after only 15 min by co-compartmentalizing single mouse hybridoma cells, a fluorescent probe and single beads coated with anti-mouse IgG antibodies in 50-pl droplets. the beads capture the secreted antibodies and, when the captured antibodies bind to the probe, the fluorescence becomes localized on the beads, generating a clearly distinguishable fluorescence signal that enables droplet sorting at ~200 Hz as well as cell enrichment. the microfluidic system described is easily adapted for screening other intracellular, cell-surface or secreted proteins and for quantifying catalytic or regulatory activities. In order to screen ~1 million cells, the microfluidic operations require 2–6 h; the entire process, including preparation of microfluidic devices and mammalian cells, requires 5–7 d.
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影响因子：
6.1
作者：
Churski, Krzysztof;Kaminski, Tomasz S.;Garstecki, Piotr
通讯作者：
Garstecki, Piotr
影响因子：
6.1
作者：
Baret, Jean-Christophe;Miller, Oliver J.;Griffiths, Andrew D.
通讯作者：
Griffiths, Andrew D.
影响因子：
3.7
作者：
Asmolov, ES
通讯作者：
Asmolov, ES
DOI：
10.1073/pnas.0704958104
发表时间：
2007-11-27
影响因子：
11.1
作者：
Di Carlo, Dino;Irimia, Daniel;Toner, Mehmet
通讯作者：
Toner, Mehmet
影响因子：
7.4
作者：
Chen, Fangyuan;Zhan, Yihong;Lu, Chang
通讯作者：
Lu, Chang