Recent advances in the use of microfluidic technologies for single cell analysis

Abstract

The inherent heterogeneity in cell populations has become of great interest and importance as analytical techniques have improved over the past decades. With the advent of personalized medicine, understanding the impact of this heterogeneity has become an important challenge for the research community. Many different microfluidic approaches with varying levels of throughput and resolution exist to study single cell activity. In this review, we take a broad view of the recent microfluidic developments in single cell analysis based on microwell, microchamber, and droplet platforms. We cover physical, chemical, and molecular biology approaches for cellular and molecular analysis including newly emerging genome-wide analysis.

Fig. 1

Microfluidics-based single-cell isolation methods. (A) U-shaped traps. Reprinted with permission from D. D. Carlo, et al., Anal. Chem., 2006, 78, 4925–4930. Copyright 2006 American Chemical Society. (B) Integrated valve traps. Reprinted with permission from J. W. Hong, et al., Nat. Biotechnol., 2004, 22, 435–439. Copyright 2004 Macmillan Publishers Ltd. (C) DEP traps. Reprinted with permission from U. C. Schroder, et al., Anal. Chem., 2013, 85, 10717–10724. Copyright 2013 American Chemical Society. (D) Optical traps. Reprinted with permission from A. Y. Lau, et al., Lab Chip, 2008, 8, 1116–1120. Copyright 2008 The Royal Society of Chemistry. (E) Droplets. Reprinted with permission from J. F. Edd, et al., Lab Chip, 2008, 8, 1262–1264. Copyright 2008 The Royal Society of Chemistry. (F) Microwells. Reprinted with permission from A. Revzin, et al., Lab Chip, 2005, 5, 30–37. Copyright 2005 The Royal Society of Chemistry.

Fig. 2

Microfluidics-based physical analysis of single cells. (A) Single-cell deformability cytometry. Reprinted with permission from H. T. K. Tse, et al., Sci. Transl. Med., 2013, 5, 212ra163–212ra163. Copyright 2013 The American Association for the Advancement of Science. (B) Single-cell impedance cytometry. Reprinted with permission from D. Holmes, et al., Lab Chip, 2009, 9, 2881–2889. Copyright 2009 The Royal Society of Chemistry.

Fig. 3

Microfluidics-based chemical analysis of single cells. (A) Chemical cytometry. Reprinted with permission from H. -Y. Wang, et al., Chem. Commun., 2006, 33, 3528–3530. Copyright 2006 The Royal Society of Chemistry. (B) TIRF (Total Internal Reflection Fluorescence) flow cytometry. Reprinted with permission from J. Wang, et al., Anal. Chem., 2008, 80, 9840–9844. Copyright 2008 American Chemical Society. (C) CARS (Coherent Anti-Stokes Raman Spectroscopy) cytometry. Reprinted with permission from H. -W. Wang, et al., Opt. Express, 2008, 16, 5782–5789. Copyright 2008 Optical Society of America. (D) Mass cytometry. Reprinted with permission from S. C. Bendall, et al., Science, 2011, 332, 687–696. Copyright 2011 The American Association for the Advancement of Science.

Fig. 4

Single cell analysis based on immunoassays and PCR. (A) Single-cell immunoassay. Reprinted with permission from Y. Lu, et al., Proc. Natl. Acad. Sci. U. S. A., 2015, 112, E607–E615. Copyright 2015 National Academy of Sciences. (B) PCR-based single-locus analysis. Reprinted with permission from D. J. Eastburn, et al., Nucleic Acids Res., 2014, 42, e128. Copyright 2014 Oxford University Press.

Fig. 5

Single-cell transcriptomic and genomic analysis. (A) Single-cell transcriptomic analysis. Reprinted with permission from E. Z. Macosko, et al., Cell, 2015, 161, 1202–1214. Copyright 2015 Elsevier. (B) Single-cell genomic analysis. Reprinted with permission from F. Lan, et al., Nat. Biotechnol., 2017, 35, 640–646. Copyright 2017 Macmillan Publishers Ltd.