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. 10The 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.

Multiplexed single cell protein expression analysis in solid tumours using a miniaturised microfluidic assay

“…We have previously developed single cell protein analysis methods and have tested and validated them on a range of cell lines [14][15][16]. These methods are based on microfluidics, with optical trap cell handling and incorporating single molecule detection.…”

Laser-based techniques for non-destructive surgery on individual cells

“…This would provide a truly quantitative read-out of cellular behavior. 10 Optical traps are also useful for injecting material. We found that cells grown on amorphous silicon (as used in solar panels) selectively and reversibly developed membrane nanopores under illumination from IR lasers, to which they would typically be transparent.…”

Optical tweezers for precise control of micro-bubble arrays: in situ temperature measurement

“…Sonoporation, or sonically enhanced permeability of cell membranes, has been demonstrated using both laser and acoustically induced microcavitation [9]. Single cell lysis via shock waves caused by laser induced microcavitation, has facilitated single cell proteomics [10].…”