High-Throughput Particle Uptake Analysis by Imaging Flow Cytometry.
High-Throughput Particle Uptake Analysis by Imaging Flow Cytometry.
复制标题
通过成像流式细胞术进行高通量颗粒摄取分析。
DOI：
10.1002/cpcy.19
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发表时间：
2017
影响因子：
--
通讯作者：
Criss,AlisonK
中科院分区：
文献类型：
--
作者：
Smirnov,Asya;Solga,MichaelD;Lannigan,Joanne;Criss,AlisonK
Quantifying the efficiency of particle uptake by host cells is important in the fields of infectious diseases, autoimmunity, cancer, developmental biology, and drug delivery. Here we present a protocol for high‐throughput analysis of particle uptake by imaging flow cytometry, using the bacteriumNeisseria gonorrhoeaeattached to and internalized by neutrophils as an example. Cells are exposed to fluorescently labeled bacteria, fixed, and stained with a bacteria‐specific antibody of a different fluorophore. Thus, in the absence of a permeabilizing agent, extracellular bacteria are double‐labeled with two fluorophores while intracellular bacteria remain single‐labeled. A spot count algorithm is used to determine the number of single‐ and double‐labeled bacteria in individual cells, to calculate the percent of cells associated with bacteria, percent of cells with internalized bacteria, and percent of cell‐associated bacteria that are internalized. These analyses quantify bacterial association and internalization across thousands of cells and can be applied to diverse experimental systems. © 2017 by John Wiley & Sons, Inc.
