A first step towards practical single cell proteomics: a microfluidic antibody capture chip with TIRF detection

Lab Chip. 2011 Apr 7;11(7):1256-61. doi: 10.1039/c0lc00613k. Epub 2011 Feb 23.

Abstract

We have developed a generic platform to undertake the analysis of protein copy number from single cells. The approach described here is 'all-optical' whereby single cells are manipulated into separate analysis chambers using an optical trap; single cells are lysed by a shock wave caused by laser-induced microcavitation, and the protein released from a single cell is measured by total internal reflection microscopy as it is bound to micro-printed antibody spots within the device. The platform was tested using GFP transfected cells and the relative precision of the measurement method was determined to be 88%. Single cell measurements were also made on a breast cancer cell line to measure the relative levels of unlabelled human tumour suppressor protein p53 using a chip incorporating an antibody sandwich assay format. These results suggest that this is a viable method for measuring relative protein levels in single cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Cell Line, Tumor
  • Humans
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Microscopy, Fluorescence / methods*
  • Proteomics / instrumentation
  • Proteomics / methods*
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*
  • Tumor Suppressor Protein p53 / analysis
  • Tumor Suppressor Protein p53 / immunology

Substances

  • Antibodies, Monoclonal
  • Tumor Suppressor Protein p53