Improved strategy for jet-in-air cell sorting with high purity, yield, viability, and genome stability.
FEBS Open Bio
; 11(9): 2453-2467, 2021 09.
Article
in En
| MEDLINE
| ID: mdl-34233080
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Flow Cytometry
Type of study:
Guideline
Limits:
Humans
Language:
En
Journal:
FEBS Open Bio
Year:
2021
Document type:
Article
Affiliation country:
China