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Review
. 2022 Jan 21;12(2):58.
doi: 10.3390/bios12020058.

Microfluidic Compartmentalization Platforms for Single Cell Analysis

Xuhao Luo  1 , Jui-Yi Chen  1 , Marzieh Ataei  2 , Abraham Lee  1   2
Affiliations
Review

Microfluidic Compartmentalization Platforms for Single Cell Analysis

Xuhao Luo et al. Biosensors (Basel). .

Abstract

Many cellular analytical technologies measure only the average response from a cell population with an assumption that a clonal population is homogenous. The ensemble measurement often masks the difference among individual cells that can lead to misinterpretation. The advent of microfluidic technology has revolutionized single-cell analysis through precise manipulation of liquid and compartmentalizing single cells in small volumes (pico- to nano-liter). Due to its advantages from miniaturization, microfluidic systems offer an array of capabilities to study genomics, transcriptomics, and proteomics of a large number of individual cells. In this regard, microfluidic systems have emerged as a powerful technology to uncover cellular heterogeneity and expand the depth and breadth of single-cell analysis. This review will focus on recent developments of three microfluidic compartmentalization platforms (microvalve, microwell, and microdroplets) that target single-cell analysis spanning from proteomics to genomics. We also compare and contrast these three microfluidic platforms and discuss their respective advantages and disadvantages in single-cell analysis.

Keywords: droplets; microvalves; microwells; single-cell analysis; single-cell compartmentalization.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The number of publications from Web of Science from the year 2000 to 2021 on microvalve, microwell, and droplet microfluidics for single-cell analysis.
Figure 2
Figure 2
Fluidigm C1 IFC Autoprep System. (A) Illustration of C1 IFC: (left) IFC with carrier; (middle) diagram of the IFC connection; (right) example of one single cell immobilized in one captured site on IFC. (B) Schematic of the C1 IFC operation. Reprinted with permission.
Figure 3
Figure 3
Schematic of DamID protocol and the function of each chamber [40]. Reprinted with permission.
Figure 4
Figure 4
Illustration of microfluidic device and the association/dissociation kinetics. (A) The configuration of the device. (B) Visualization of the inlets (yellow and green) and the control channels (red). The image at the bottom right shows the beads captured by the sieve valves. (C) Measurement of association rate. (D) Measurement of dissociation rate [48]. Reprinted with permission.
Figure 5
Figure 5
Schematic of (A) scRNA printed on a glass slide and (B) captured on a barcoded bead [68]. Reprinted with permission.
Figure 6
Figure 6
Techniques of capturing the antibodies secreted by single cells in microwells. (A) single cells are captured in microwells. (B) Microwells are sealed with 2nd antibody-coated or antigen-coated glass slide. (C) The presence of the target antibody is visualized by fluorescently labeled antigen or secondary antibody. (D,E) Functionalization of the well. (D) Functionalize the top surface of the well, as reported by Muraguchi et. al. [54,59]. (E) Functionalize the top and the inner surface of the well, as studied by Torres et al. [79].
Figure 7
Figure 7
Illustration of antibody-secreting cells using VyCAP. The microwells have a 5 µm hole at the bottom. The wells are brought in contact with a ligand-coated membrane (gray) and the antibodies were captured on the membrane. The captured antibodies were visualized by fluorescent immunostaining [83]. Reprinted with permission.
Figure 8
Figure 8
In situ single-cell Western blot (in situ scWB). (A) Workflow of in situ scWB illustrating an array from one of the 2000 arrays on a device. (B) Preparation of FN-functionalized PA microwell [85]. Reprinted with permission.
Figure 9
Figure 9
Schematic illustration of the configuration of droplet based microfluidic platform. The center stream contains a mixture of microspheres, cells, and secondary antibodies (mix), while the two opposing side streams contain the oil phase. The encapsulated cells proceed into the downstream incubation region for cell analysis and can then be sorted based on secretion of the interrogated analyte [102]. Reprinted with permission.
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