title: A first step towards practical single cell proteomics: a microfluidic antibody capture chip with TIRF detection.
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.
authors.affiliation: Single Cell Proteomics Project, Institute of Chemical Biology, Imperial College London, Exhibition Road, London SW7 2AZ, UK. ali.salehi-reyhani@imperial.ac.uk
authors.forename: Ali
authors.initials: A
authors.lastname: Salehi-Reyhani
authors.forename: Joseph
authors.initials: J
authors.lastname: Kaplinsky
authors.forename: Edward
authors.initials: E
authors.lastname: Burgin
authors.forename: Miroslava
authors.initials: M
authors.lastname: Novakova
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: deMello
authors.forename: Richard H
authors.initials: RH
authors.lastname: Templer
authors.forename: Peter
authors.initials: P
authors.lastname: Parker
authors.forename: Mark A A
authors.initials: MA
authors.lastname: Neil
authors.forename: Oscar
authors.initials: O
authors.lastname: Ces
authors.forename: Paul
authors.initials: P
authors.lastname: French
authors.forename: Keith R
authors.initials: KR
authors.lastname: Willison
authors.forename: David
authors.initials: D
authors.lastname: Klug
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D045744:Cell Line, Tumor; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D016159:Tumor Suppressor Protein p53