Droplet Microfluidic Flow Cytometer For Sorting On Transient Cellular Responses Of Genetically-Encoded Sensors.

Droplet Microfluidic Flow Cytometer For Sorting On Transient Cellular Responses Of Genetically-Encoded Sensors.
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液滴微流体流式细胞仪,用于对遗传编码传感器的瞬时细胞反应进行分类。

DOI:
10.1021/acs.analchem.6b03235
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发表时间:
2017-01-03
影响因子:
7.4
通讯作者:
Jimenez R
Jimenez R
中科院分区:
化学1区
文献类型:
--
作者:
Fiedler BL;Van Buskirk S;Carter KP;Qin Y;Carpenter MC;Palmer AE;Jimenez R

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荧光生物传感器是重要的测量工具,用于在体内定量pH值,金属离子浓度和其他分析物,以及物理参数,如膜电位。这些传感器的发展及其在检查细胞异质性方面的实施都需要基于化学或物理扰动前后的荧光水平来测量和分选细胞的技术。我们开发了一个液滴微流控平台,用于在添加外源分析物后表达的荧光生物传感器的体内反应的基础上筛选和分离细胞群。我们展示了在几秒钟的时间点范围内解决两个遗传编码的Zn2+传感器的响应的能力,并随后以高精度对不同比例的混合细胞群体进行排序。
Fluorescent biosensors are important measurement tools for in vivo quantification of pH, concentrations of metal ions and other analytes, and physical parameters such as membrane potential. Both the development of these sensors and their implementation in examining cellular heterogeneity requires technology for measuring and sorting cells based on the fluorescence levels before and after chemical or physical perturbations. We developed a droplet microfluidic platform for the screening and separation of cell populations on the basis of the in vivo response of expressed fluorescence-based biosensors after addition of an exogenous analyte. We demonstrate the capability to resolve the responses of two genetically-encoded Zn2+ sensors at a range of time points spanning several seconds and subsequently sort a mixed-cell population of varying ratios with high accuracy.
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