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Separation-encoded microparticles for single-cell western blotting

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Abstract

Direct measurement of proteins from single cells has been realized at the microscale using microfluidic channels, capillaries, and semi-enclosed microwell arrays. Although powerful, these formats are constrained, with the enclosed geometries proving cumbersome for multistage assays, including electrophoresis followed by immunoprobing. We introduce a hybrid microfluidic format that toggles between a planar microwell array and a suspension of microparticles. The planar array is stippled in a thin sheet of polyacrylamide gel, for efficient single-cell isolation and protein electrophoresis of hundreds-to-thousands of cells. Upon mechanical release, array elements become a suspension of separation-encoded microparticles for more efficient immunoprobing due to enhanced mass transfer. Dehydrating microparticles offer improved analytical sensitivity owing to in-gel concentration of fluorescence signal for high-throughput single-cell targeted proteomics.

Graphical abstract: Separation-encoded microparticles for single-cell western blotting

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Publication details

The article was received on 14 Sep 2019, accepted on 14 Nov 2019 and first published on 15 Nov 2019


Article type: Paper
DOI: 10.1039/C9LC00917E
Lab Chip, 2020, Advance Article

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    Separation-encoded microparticles for single-cell western blotting

    B. Gumuscu and A. E. Herr, Lab Chip, 2020, Advance Article , DOI: 10.1039/C9LC00917E

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