Abstract
Many conventional biochemical assays are performed using populations of cells to determine their quantitative biomolecular profiles. However, population averages do not reflect actual physiological processes in individual cells, which occur either on short time scales or nonsynchronously. Therefore, accurate analysis at the single-cell level has become a highly attractive tool for investigating cellular content. Microfluidic chips with arrays of microwells were developed for single-cell chemical lysis in the present study. The cellular occupancy in 30-μm-diameter microwells (91.45%) was higher than that in 20-μm-diameter microwells (83.19%) at an injection flow rate of 2.8 μL/min. However, most of the occupied 20-μm-diameter microwells contained individual cells. The results of chemical lysis experiments at the single-cell level indicate that cell membranes were gradually lysed as the lysis buffer was injected; they were fully lysed after 12 s. Single-cell chemical lysis was demonstrated in the proposed microfluidic chip, which is suitable for high-throughput cell lysis.
Highlights
Cell lysis is crucial in the analysis of intracellular components containing information about genetic or disease characteristics in genomics, proteomics, and metabolomics [1]
Phenylmethylsulfonyl fluoride (PMSF) was employed to lyse single cells deposited in the microwells
Micropatterned HeLa cells in the microfluidic chips with 20- and 30-μm-diameter microwells are shown in Figure 2; the injection flow rate of the cell sample is 2.8 μL/min
Summary
Cell lysis is crucial in the analysis of intracellular components containing information about genetic or disease characteristics in genomics, proteomics, and metabolomics [1]. Chung et al [17] proposed a microfluidic platform utilizing the hydrodynamic flow in conjunction with a careful disposition of the cell traps in an array formed by a serpentine channel for single-cell capture, stimulation, and imaging These methods have merits when existing cell assays are integrated onto a microchip platform. Various optical [27], mechanical [28], electric [29], and chemical [30] approaches for cell lysis have been proposed [31] He et al [32] proposed a method that combines optical trapping and microfluidic-based droplet generation for encapsulating single cells within a picolitre-size aqueous droplet. Microfluidic chips with microwells, 20 and 30 μm in diameter, are developed for cellular patterning using a flow method and the feasibility of chemical lysis for human carcinoma cells (HeLa cells) at the single-cell level is demonstrated. The effects of parameters such as the size of the microwells and the injection flow rate on cellular occupancy are investigated
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