Method for Sorting High-Viability Cells Based on Forward Scatter Light Width Parameter

To eliminate the interference of dead cells in samples, specific dyes are commonly used prior to cell sorting to exclude dead cells or label viable cells. However, such methods require specialized reagents, impose certain technical requirements, and increase sample preparation time.

Using jet-in-air sorters, this study found that the forward scatter width (FSC-W) signals of all singlet cells can be divided into two populations: medium FSC-W intensity (FSC-W-medium) and low FSC-W intensity (FSC-W-low). Moreover, the FSC-W-medium population exhibited significantly higher viability and lower autofluorescence. Across eight commonly used cell lines, the proportion of dead cells and autofluorescent cells in the FSC-W-medium population was significantly lower than those in the FSC-W-low population.Furthermore, quantitative proteomic analysis revealed differences in protein expression profiles between these two populations, and the FSC-W-medium cell population exhibited lower intercellular heterogeneity.