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

Autor: Yanwei Li, Yue-Ting Xing, Chun Guo, Lin-Lin Wang, Ying-Ying Huang, Hu Hu, Lintao Xu, Jiajia Wang, Xinghui Song
Rok vydání: 2021
Předmět:
Zdroj: FEBS Open Bio, Vol 11, Iss 9, Pp 2453-2467 (2021)
FEBS Open Bio
ISSN: 2211-5463
DOI: 10.1002/2211-5463.13248
Popis: 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.
Flow cytometric sorting is a vital method in biological research and clinical diagnostics. Obtaining high‐quality sorted cells is an ongoing problem. Here, we demonstrate that a certain concentration of FBS helps to increase cell yield and cell viability and keep gene expression stable. Our work provides an effective sorting strategy.
Databáze: OpenAIRE
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