Abstract
In a forest of a hundred thousand trees, no two leaves are alike. Similarly, no two cells in a genetically identical group are the same. This heterogeneity at the single-cell level has been recognized to be vital for the correct interpretation of diagnostic and therapeutic results of diseases, but has been masked for a long time by studying average responses from a population. To comprehensively understand cell heterogeneity, diverse manipulation and comprehensive analysis of cells at the single-cell level are demanded. However, using traditional biological tools, such as petri-dishes and well-plates, is technically challengeable for manipulating and analyzing single-cells with small size and low concentration of target biomolecules. With the development of microfluidics, which is a technology of manipulating and controlling fluids in the range of micro- to pico-liters in networks of channels with dimensions from tens to hundreds of microns, single-cell study has been blooming for almost two decades. Comparing to conventional petri-dish or well-plate experiments, microfluidic single-cell analysis offers advantages of higher throughput, smaller sample volume, automatic sample processing, and lower contamination risk, etc., which made microfluidics an ideal technology for conducting statically meaningful single-cell research. In this review, we will summarize the advances of microfluidics for single-cell manipulation and analysis from the aspects of methods and applications. First, various methods, such as hydrodynamic and electrical approaches, for microfluidic single-cell manipulation will be summarized. Second, single-cell analysis ranging from cellular to genetic level by using microfluidic technology is summarized. Last, we will also discuss the advantages and disadvantages of various microfluidic methods for single-cell manipulation, and then outlook the trend of microfluidic single-cell analysis.
Highlights
Over the past few decades, cellular heterogeneity has gradually been emphasized on fundamental biological and clinical research as numerous novel tools/methods for single-cell analysis have emerged [1]
We mainly focus on microfluidic technologies for single-cell manipulation analysis from the aspects of methods and applications
Hu et al used a near-infrared laser focused on indium tin oxide (ITO) glass to generate thermocapillary effect that can trap and transport living single cells with forces of up to 40 pN [77]
Summary
Over the past few decades, cellular heterogeneity has gradually been emphasized on fundamental biological and clinical research as numerous novel tools/methods for single-cell analysis have emerged [1]. Taking flow cytometry as an example, they are automatic, capable of multiple detections, and efficient in single-cell sorting, they are bulky, mechanically complicated, expensive, and demanding for relatively large sample volumes They can only be used for analyzing cells at one time-point. Many single-cell studies require single-cell capture/isolation, and different microfluidic methods, such as hydrodynamic [11,18,19], electrical [20], optical [21], magnetic [22], and acoustic [23] methods, have been developed. Various detection methods, such as fluorescence microscopy, fluorometry and mass spectroscopy, can be combined with microfluidic systems for single-cell analysis from cell morphology to secreted proteins. We discuss the technology and application trend for microfluidics based single-cell analysis
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