Issue 34, 2021

Droplet flow cytometry for single-cell analysis


The interrogation of single cells has revolutionised biology and medicine by providing crucial unparalleled insights into cell-to-cell heterogeneity. Flow cytometry (including fluorescence-activated cell sorting) is one of the most versatile and high-throughput approaches for single-cell analysis by detecting multiple fluorescence parameters of individual cells in aqueous suspension as they flow past through a focus of excitation lasers. However, this approach relies on the expression of cell surface and intracellular biomarkers, which inevitably lacks spatial and temporal phenotypes and activities of cells, such as secreted proteins, extracellular metabolite production, and proliferation. Droplet microfluidics has recently emerged as a powerful tool for the encapsulation and manipulation of thousands to millions of individual cells within pico-litre microdroplets. Integrating flow cytometry with microdroplet architectures surrounded by aqueous solutions (e.g., water-in-oil-in-water (W/O/W) double emulsion and hydrogel droplets) opens avenues for new cellular assays linking cell phenotypes to genotypes at the single-cell level. In this review, we discuss the capabilities and applications of droplet flow cytometry (DFC). This unique technique uses standard commercially available flow cytometry instruments to characterise or select individual microdroplets containing single cells of interest. We explore current challenges associated with DFC and present our visions for future development.

Graphical abstract: Droplet flow cytometry for single-cell analysis

Article information

Article type
Review Article
04 Apr 2021
06 Jun 2021
First published
14 Jun 2021
This article is Open Access
Creative Commons BY license

RSC Adv., 2021,11, 20944-20960

Droplet flow cytometry for single-cell analysis

M. Li, H. Liu, S. Zhuang and K. Goda, RSC Adv., 2021, 11, 20944 DOI: 10.1039/D1RA02636D

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