Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2016 Jul 5;21(7):881.
doi: 10.3390/molecules21070881.

Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

Na Wen  1 , Zhan Zhao  2 , Beiyuan Fan  3 , Deyong Chen  4 , Dong Men  5 , Junbo Wang  6 , Jian Chen  7
Affiliations
Review

Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

Na Wen et al. Molecules. .

Abstract

This article reviews recent developments in droplet microfluidics enabling high-throughput single-cell analysis. Five key aspects in this field are included in this review: (1) prototype demonstration of single-cell encapsulation in microfluidic droplets; (2) technical improvements of single-cell encapsulation in microfluidic droplets; (3) microfluidic droplets enabling single-cell proteomic analysis; (4) microfluidic droplets enabling single-cell genomic analysis; and (5) integrated microfluidic droplet systems enabling single-cell screening. We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on key performances of throughput, multifunctionality, and absolute quantification.

Keywords: droplet microfluidics; high-throughput; single-cell encapsulation; single-cell genetic analysis; single-cell proteomic analysis; single-cell screening.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(a) Schematic of a microfluidic T channel and (b) sequences of images showing the encapsulation of a single B lymphocyte into an aqueous droplet in silicone oil. Optical trapping was used to transport and position the cell close to the water/oil interface. Upon application of a pressure pulse to the microchannels, the cell was carried away by the flow as the droplet was sheared off. Reproduction with permission from [34].
Figure 2
Figure 2
Size-based droplet sorting after cellular encapsulation. A purely hydrodynamic approach for single-cell encapsulation, followed by spontaneous self-sorting of these droplets based on lateral drift of deformable objects in a shear flow, and sterically-driven dispersion in a compressional flow. Reproduction with permission from [37].
Figure 3
Figure 3
Inertial flow-based cell spacing and single-cell encapsulation using (a) a high aspect-ratio straight microchannel (reproduction with permission from [41]); (b) a curved microchannel (reproduction with permission from [42]); and (c) a short pinched flow channel (reproduction with permission from [43]).
Figure 4
Figure 4
Microfluidic droplets enabling single-cell proteomic analysis. (a) Individual E coli and substrate 3-O-methylfluorescein-phosphates were encapsulated within single droplets where the substrates were enzymatically hydrolyzed by the target enzyme alkaline phosphatase expressed by E coli, leading to fluorescent detections. Reproduction with permission from [47]; (b) Both microspheres conjugated with capture antibodies and detection fluorescence labeled antibodies were encapsulated with single cells and the secreted IL-10 of CD4 + CD25 + regulatory T cells was captured on the microsphere surface and detected via detection antibodies, generating localized fluorescent signals on microsphere surfaces. Reproduction with permission from [58].
Figure 5
Figure 5
Microfluidic droplets enabling single-cell genomic analysis. (a) Individual cells together with primer-functionalized microbeads were encapsulated in uniform PCR mix droplets. After bulk PCR amplification, the droplets were lysed and the beads were recovered and rapidly analyzed via flow cytometry. Reproduction with permission from [71]; and (b) an agarose droplet-based microfluidic method for emulsification RT-PCR, where reverse primers were covalently conjugated to agarose, which functioned as the trapping matrix to replace conventional primer functionalized microbeads, resulting in high PCR efficiency (~95%). Reproduction with permission from [81].
Figure 6
Figure 6
Integrated microfluidic system for single-cell screening, including key steps of cell encapsulation, incubation, fluorescence detection of metabolic molecules, and droplet sorting relying on the fluorescent intensities. Reproduction with permission from [91].
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Wang D., Bodovitz S. Single cell analysis: The new frontier in ‘omics’. Trends Biotechnol. 2010;28:281–290. doi: 10.1016/j.tibtech.2010.03.002. - DOI - PMC - PubMed
    1. Fritzsch F.S., Dusny C., Frick O., Schmid A. Single-cell analysis in biotechnology, systems biology, and biocatalysis. Annu. Rev. Chem. Biomol. Eng. 2012;3:129–155. doi: 10.1146/annurev-chembioeng-062011-081056. - DOI - PubMed
    1. Tsioris K., Torres A.J., Douce T.B., Love J.C. A new toolbox for assessing single cells. Annu. Rev. Chem. Biomol. Eng. 2014;5:455–477. doi: 10.1146/annurev-chembioeng-060713-035958. - DOI - PMC - PubMed
    1. Vasdekis A.E., Stephanopoulos G. Review of methods to probe single cell metabolism and bioenergetics. Metab. Eng. 2015;27:115–135. doi: 10.1016/j.ymben.2014.09.007. - DOI - PMC - PubMed
    1. Klepárník K., Foret F. Recent advances in the development of single cell analysis—A review. Anal. Chim. Acta. 2013;800:12–21. doi: 10.1016/j.aca.2013.09.004. - DOI - PubMed