Abstract
The inherent heterogeneity of individual cells in cell populations plays significant roles in disease development and progression, which is critical for disease diagnosis and treatment. Substantial evidences show that the majority of traditional gene profiling methods mask the difference of individual cells. Single cell sequencing can provide data to characterize the inherent heterogeneity of individual cells, and reveal complex and rare cell populations. Different microfluidic technologies have emerged for single cell researches and become the frontiers and hot topics over the past decade. In this review article, we introduce the processes of single cell sequencing, and review the principles of microfluidics for single cell analysis. Also, we discuss the common high-throughput single cell sequencing technologies along with their advantages and disadvantages. Lastly, microfluidics applications in single cell sequencing technology for the diagnosis of cancers and immune system diseases are briefly illustrated.
Highlights
A single cell is the basic structural and functional unit of living organisms
We summarize the principles of microfluidics for single cell analysis, such as traps-based, vavles-based, and droplet-based methods
A great deal of microfluidic devices have been proposed for single cell analysis in the past years [30] and we focus on the ability of the device on isolation of individual cells for further downstream analysis and culture
Summary
A single cell is the basic structural and functional unit of living organisms. Cells derived from the same type of cells under the same external stimulus or physiological conditions may exhibit cell-to-cell differences [1]. Single cell analysis based on multi-technical combinational microfluidics has revolutionized plenty of research fields, and its application on single cell sequencing has received broad attention for its high-throughput characteristics. The approaches of separating single cell samples include limiting dilution, micromanipulation, laser capture microdissection (LCM), fluorescence activated cell sorting (FACS) and microfluidic technology. Limiting dilution mainly uses hand-pipettes or pipetting robots to isolate individual cells, whose probability for the number of cells per aliquot obey Poisson’s distribution This method is obsolete because it cannot exclude some small cell populations [25]. LCM is an advanced tool to quickly and precisely acquire single cells or cell compartments from mostly solid tissue samples after slice and dyeing treatment This method successfully maintains cell morphology and structure and preserves the spatial location information [27].
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