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Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability

Authors :
Xinghui Song
Jiajia Wang
Yanwei Li
Yueting Xing
Chun Guo
Yingying Huang
Lintao Xu
Hu Hu
Lin‐lin Wang
Source :
FEBS Open Bio, Vol 11, Iss 9, Pp 2453-2467 (2021)
Publication Year :
2021
Publisher :
Wiley, 2021.

Abstract

Flow cytometric sorting is a vital tool in biological research and clinical diagnostics. Theoretically, a high‐speed jet‐in‐air sorter is a fluorescent‐activated cell sorting sorter that ideally processes cells with high purity, yield, and viability. However, high‐speed jet‐in‐air sorting is a complex process due to its inherent requirements for high fluidic stability and electronic and timing precision. Here, we report that an additional manual correction of drop delay leads to improved cell yield. Adding 2% FBS to the loading buffer had no significant effect on the fate of sorted cells in 4 h. However, the addition of a suitable concentration of FBS/BSA in the collecting buffer resulted in a notable increase in cell count and proliferation and a significant decrease in cell apoptosis for cell lines and primary cells. Moreover, the level of gene expression remained steady in the 5% FBS collecting buffer. In summary, here we demonstrate techniques that can be easily followed to refine sorted yields of healthy cells.

Details

Language :
English
ISSN :
22115463
Volume :
11
Issue :
9
Database :
Directory of Open Access Journals
Journal :
FEBS Open Bio
Publication Type :
Academic Journal
Accession number :
edsdoj.846f0847113f4c96b25019c0696ae22b
Document Type :
article
Full Text :
https://doi.org/10.1002/2211-5463.13248