Improving Single-Cell Encapsulation Efficiency and Reliability through Neutral Buoyancy of Suspension.

Hangrui Liu,Ming Li,Yan Wang,Lianmei Jiang,Jim Piper

doi:10.3390/mi11010094

Abstract

Single-cell analysis is of critical importance in revealing cell-to-cell heterogeneity by characterizing individual cells and identifying minority sub-populations of interest. Droplet-based microfluidics has been widely used in the past decade to achieve high-throughput single-cell analysis. However, to maximize the proportion of single-cell emulsification is challenging due to cell sedimentation and aggregation. The purpose of this study was to investigate the influence of single-cell encapsulation and incubation through the use of neutral buoyancy. As a proof of concept, OptiPrep™ was used to create neutrally buoyant cell suspensions of THP-1, a human monocytic leukemia cell line, for single-cell encapsulation and incubation. We found that using a neutrally buoyant suspension greatly increased the efficiency of single-cell encapsulation in microdroplets and eliminated unnecessary cell loss. Moreover, the presence of OptiPrep™ was shown to not affect cellular viability. This method significantly improved the effectiveness of single-cell study in a non-toxic environment and is expected to broadly facilitate single-cell analysis.

Highlights

Single-cell analysis has been attracting great interest from both academia and industry as a powerful technique in the field of medical diagnosis, tissue engineering, and cell biology
We developed a medium density matching strategy to improve single-cell encapsulation efficiency and reliability through neutral buoyancy of cell suspension as shown in Micromachines 2020, 11, x in the non-density matched suspension
We alleviated the negative influence of gravity by adding OptiPrepTM to achieve near neutral buoyancy within the suspension

Summary

Introduction

Single-cell analysis has been attracting great interest from both academia and industry as a powerful technique in the field of medical diagnosis, tissue engineering, and cell biology. Individual cells of interest can be sorted from the majority of the population for further studies. This technology has been demonstrated as a powerful tool to investigate cancer biology, by profiling marked heterogeneity, development of new diagnostics, and personalized medicine. Traditional flow cytometry is mostly used in single-cell analysis to scrutinize heterogeneous cell populations in a high throughput and multiplexing manner [3,4]. Flow cytometry allows only single time point measurements, the same single cells can neither be observed over long periods nor suitable for repetitive measurements [2,5]

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