Second, it allows the sorting of a large range of droplet volumes: from 20 fL 12 to 10 nL 13. First, dielectrophoresis is very efficient for the sorting of water-in-oil droplets, as it is mainly governed by the dielectric contrast between water and oil, independently of additives in these phases. The former approach has indeed been democratized for several reasons. However, the most widespread method is dielectrophoretic-based sorting. A high viability of encapsulated cells in such droplets has been observed over several days 10, and various active droplet sorting solutions such as acoustic, magnetic, pneumatic, thermal, and electric actuation have already been described 11. Even though intracellular staining flow cytometry has been described, the use of protein transport inhibitors can interfere with the analysis 9.Īn alternative solution consists in the compartmentalization of cells in monodisperse emulsion droplets. Whereas both FACS and the latter systems are efficient to screen compounds remaining within the cell or on its surface, they are not suited for cytoplasmic or secreted proteins screening.
A large variety of such microdevices has been developed in recent years, based on different physical mechanisms such as optical manipulation 5, mechanical systems 6, acoustophoresis 7, and electrokinetics 8. Compared with FACS, fluorescence-based cell sorting microsystems allow to reduce sample amounts to eliminate potentially biohazardous aerosols 4 and to implement complex assays. Moreover, it is not compatible with the analysis of small cell populations (< 10 5 cells 3). However, a main drawback of FACS is that it cannot support real-time analysis of single cell or integration of complex assays involving single-cell manipulation, treatment, and final detection 2. The gold standard technology for this purpose is fluorescence-activated cell sorting (FACS) 1.
Fluorescence-based cell sorting is essential in numerous biological assays requiring high-throughput analysis and sorting of single cells.
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