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一种用于高精度单细胞分选和分配的荧光激活单液滴分配器。

A Fluorescence-Activated Single-Droplet Dispenser for High Accuracy Single-Droplet and Single-Cell Sorting and Dispensing.

机构信息

Department of Chemistry , University of Washington , Seattle , Washington 98195 , United States.

Clinical Research Division , Fred Hutchinson Cancer Research Center , Seattle , Washington 98109 , United States.

出版信息

Anal Chem. 2019 May 21;91(10):6815-6819. doi: 10.1021/acs.analchem.9b01017. Epub 2019 May 9.

Abstract

The ability to sort and dispense droplets accurately is essential to droplet-based single-cell analysis. Here, we describe a fluorescence-activated single-droplet dispenser (FASD) that is analogous to a conventional fluorescence-activated cell sorter, but sorts droplets containing single cells within an oil emulsion. The FASD system uses cytometric detection and electrohydrodynamic actuation-based single-droplet manipulation, allowing droplet isolation and dispensing with high efficiency and accuracy. The system is compatible with multiwell plates and can be integrated with existing microfluidic devices and large-scale screening systems. By enabling sorting based on single-cell reactions such as PCR, this platform will help expand the basis of cell sorting from mainly protein biomarkers to nucleic acid sequences and secreted biomolecules.

摘要

准确的分选和分配液滴的能力对于基于液滴的单细胞分析至关重要。在这里,我们描述了一种荧光激活的单液滴分配器(FASD),它类似于传统的荧光激活细胞分选器,但可对包含油乳液中单细胞的液滴进行分选。FASD 系统使用基于细胞计检测和基于电动液压的单液滴操作,可实现高效、准确的液滴分离和分配。该系统与多孔板兼容,可与现有的微流控设备和大规模筛选系统集成。通过使基于单细胞反应(如 PCR)的分选成为可能,该平台将有助于将细胞分选的基础从主要的蛋白质生物标志物扩展到核酸序列和分泌生物分子。

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Single-Cell RT-PCR in Microfluidic Droplets with Integrated Chemical Lysis.微流控液滴中的单细胞 RT-PCR 与集成化学裂解
Anal Chem. 2018 Jan 16;90(2):1273-1279. doi: 10.1021/acs.analchem.7b04050. Epub 2018 Jan 3.
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