Separation-encoded microparticles for single-cell western blotting

Lab Chip. 2020 Jan 7;20(1):64-73. doi: 10.1039/c9lc00917e. Epub 2019 Nov 27.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylic Resins / chemistry
  • Blotting, Western*
  • Cells, Cultured
  • Estrogen Receptor alpha / chemistry
  • Estrogen Receptor alpha / isolation & purification*
  • Gels / chemistry
  • HEK293 Cells
  • Humans
  • Lab-On-A-Chip Devices*
  • MCF-7 Cells
  • Proteomics
  • Single-Cell Analysis*

Substances

  • Acrylic Resins
  • Estrogen Receptor alpha
  • Gels
  • polyacrylamide