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

Xinghui Song,Chun Guo,Hu Hu,Jiajia Wang,Yingying Huang,Yanwei Li,Lin‐Lin Wang,Yueting Xing,Lintao Xu

doi:10.1002/2211-5463.13248

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.

Highlights

The enrichment of viable cells is an essential step in single-cell sequencing for clinical and biotechnology research applications
We investigated the effects on cell count, apoptosis, proliferation and endogenous gene expression caused by adding different concentrations of fetal bovine serum (FBS) or bovine serum albumin (BSA) to the loading buffer and the collecting buffer during fluorescent-activated cell sorting (FACS)
The result was that the sort yield was averagely 88.4% according to the automatic drop delay value, and the yield reached averagely 99.81% when the drop delay value was modified by the manual method under the same conditions (Fig 1.A)

Summary

Introduction

The enrichment of viable cells is an essential step in single-cell sequencing for clinical and biotechnology research applications. A high-end jet-in-air sorter is a fluorescent-activated cell sorting sorter that ideally processes cells with high purity, yield and viability. Flow cytometric sorting is a vital tool in biological research and clinical diagnostics [1]. This method functions as a technology for isolating target cells but can provide abundant information on targeted cells, which can help to guide subsequent experiments [2,3,4]. A high-end jet-in-air sorter is a fluorescent-activated cell sorting (FACS) sorter that ideally processes cells with high purity, yield and viability [6]. A highly accurate sorting strategy to achieve high yield and good biocompatibility, including maintaining high viability and functionality, is a considerable challenge facing high-end jet-in-air sorters

Methods

Results

Discussion

Conclusion