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

Ali Salehi-Reyhani, Joseph Kaplinsky, Edward Burgin, Miroslava Novakova, Andrew J. deMello, Richard H. Templer, Peter Parker, Mark A. A. Neil, Oscar Ces, Paul French, Keith R. Willison and David Klug
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.

DOI