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

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 compl...

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Autores principales: Song, Xinghui, Wang, Jiajia, Li, Yanwei, Xing, Yueting, Guo, Chun, Huang, Yingying, Xu, Lintao, Hu, Hu, Wang, Lin‐lin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8409286/
https://www.ncbi.nlm.nih.gov/pubmed/34233080
http://dx.doi.org/10.1002/2211-5463.13248
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author Song, Xinghui
Wang, Jiajia
Li, Yanwei
Xing, Yueting
Guo, Chun
Huang, Yingying
Xu, Lintao
Hu, Hu
Wang, Lin‐lin
author_facet Song, Xinghui
Wang, Jiajia
Li, Yanwei
Xing, Yueting
Guo, Chun
Huang, Yingying
Xu, Lintao
Hu, Hu
Wang, Lin‐lin
author_sort Song, Xinghui
collection PubMed
description 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.
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spelling pubmed-84092862021-09-03 Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability Song, Xinghui Wang, Jiajia Li, Yanwei Xing, Yueting Guo, Chun Huang, Yingying Xu, Lintao Hu, Hu Wang, Lin‐lin FEBS Open Bio Methods 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. John Wiley and Sons Inc. 2021-07-30 /pmc/articles/PMC8409286/ /pubmed/34233080 http://dx.doi.org/10.1002/2211-5463.13248 Text en © 2021 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods
Song, Xinghui
Wang, Jiajia
Li, Yanwei
Xing, Yueting
Guo, Chun
Huang, Yingying
Xu, Lintao
Hu, Hu
Wang, Lin‐lin
Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title_full Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title_fullStr Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title_full_unstemmed Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title_short Improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
title_sort improved strategy for jet‐in‐air cell sorting with high purity, yield, viability, and genome stability
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8409286/
https://www.ncbi.nlm.nih.gov/pubmed/34233080
http://dx.doi.org/10.1002/2211-5463.13248
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