Microfluidic techniques for high throughput single cell analysis

Abstract

The microfabrication of microfluidic control systems and the development of increasingly sensitive molecular amplification tools have enabled the miniaturization of single cells analytical platforms. Only recently has the throughput of these platforms increased to a level at which populations can be screened at the single cell level. Techniques based upon both active and passive manipulation are now capable of discriminating between single cell phenotypes for sorting, diagnostic or prognostic applications in a variety of clinical scenarios. The introduction of multiphase microfluidics enables the segmentation of single cells into biochemically discrete picoliter environments. The combination of these techniques are enabling a class of single cell analytical platforms within great potential for data driven biomedicine, genomics and transcriptomics.

Figure 1

Microfluidic technologies have dramatically increased in throughput capacity while enabling high-purity single cell recognition or separation from bulk samples. Microfluidic techniques that have enabled the evolution of single cell analyses, and their predecessors are illustrated: a) Atomic force microscopy for biophysical marker identification[13] b) Optical stretching for single cell mechanical property measurements[50]. c) Hydrodynamic cell isolation array for single cell capture and analyses[–15]. d) Microfluidic large scale integration[3]. e) Hydrodynamic stretching for single cell mechanophenotyping[48].

Figure 2

Overview of inertial focusing and its application in various high throughput biological sample processing for single cell analyses. a) Inertial focusing behavior in microchannels with rectangular cross-section. b) High throughput inertial focusing mediated magnetophoresis[6]. c) Size based cell sorting in spiral microchannels with rectangular cross section. d) Single cell hydrodynamic stretching for mechanical phenotyping[48]. e) Deterministic cell encapsulation for precise modulation of single cell environments[53].

Figure 3

Overview of microfluidic single cell encapsulation and droplet microenvironment manipulations for downstream molecular analyses. a) Droplet single copy PCR amplification[55]. b) Droplet whole genome PCR amplification[56]. c) Droplet barcoding for single cell transcriptomics[63].