Abstract
Single-cell analysis of cell phenotypic information such as surface protein expression and nucleic acid content is essential for understanding heterogeneity within cell populations. Here the design and use of a dielectrophoresis-assisted self-digitization (SD) microfluidics chip is described; it captures single cells in isolated microchambers with high efficiency for single-cell analysis. The self-digitization chip spontaneously partitions aqueous solution into microchambers through a combination of fluidic forces, interfacial tension, and channel geometry. Single cells are guided to and trapped at the entrances of microchambers by dielectrophoresis (DEP) due to local electric field maxima created by an externally applied AC voltage. Excess cells are flushed away, and trapped cells are released into the chambers and prepared for in situ analysis by turning off the external voltage, by running reaction buffer through the chip, and by sealing the chambers with a flow of an immiscible oil phase through the surrounding channels. The use of this device in single-cell analysis is demonstrated by performing single-cell nucleic acid quantitation based on loop-mediated isothermal amplification (LAMP). This platform provides a powerful new tool for single-cell research pertaining to drug discovery. For example, the single-cell genoty** of cancer-related mutant gene observed from the digital chip could be useful biomarker for targeted therapy.
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Acknowledgments
This work was supported by NIH grant (R21EB018831 and UG3CA211139), NNSFC grant (31901056), and JSAP grant (06200053).
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Qin, Y., Wu, L., Chiu, D.T. (2023). Dielectrophoresis-Assisted Self-Digitization Chip for High-Efficiency Single-Cell Analysis. In: Li, P.C., Wu, A.R. (eds) Single-Cell Assays. Methods in Molecular Biology, vol 2689. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3323-6_3
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DOI: https://doi.org/10.1007/978-1-0716-3323-6_3
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