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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.

Original publication

DOI

10.1039/c0lc00613k

Type

Journal article

Journal

Lab Chip

Publication Date

07/04/2011

Volume

11

Pages

1256 - 1261

Keywords

Animals, Antibodies, Monoclonal, Cell Line, Tumor, Humans, Microfluidic Analytical Techniques, Microscopy, Fluorescence, Proteomics, Single-Cell Analysis, Tumor Suppressor Protein p53