title: Rapid quantification of live cell receptors using bioluminescence in a flow-based microfluidic device.
abstract: The number of receptors expressed by cells plays an important role in controlling cell signaling events, thus determining its behaviour, state and fate. Current methods of quantifying receptors on cells are either laborious or do not maintain the cells in their native form. Here, a method integrating highly sensitive bioluminescence, high precision microfluidics and small footprint of lensfree optics is developed to quantify cell surface receptors. This method is safe to use, less laborious, and faster than the conventional radiolabelling and near field scanning methods. It is also more sensitive than fluorescence based assays and is ideal for high throughput screening. In quantifying β(1) adrenergic receptors expressed on the surface of H9c2 cardiomyocytes, this method yields receptor numbers from 3.12 × 10(5) to 9.36 × 10(5) receptors/cell which are comparable with current methods. This can serve as a very good platform for rapid quantification of receptor numbers in ligand/drug binding and receptor characterization studies, which is an important part of pharmaceutical and biological research.
authors.affiliation: Department of Biomedical Engineering, 9 Engineering Drive 1, Block EA, #03-12, National University of Singapore, Singapore, 117575.
authors.forename: Ramesh
authors.initials: R
authors.lastname: Ramji
authors.forename: Cheong Fook
authors.initials: CF
authors.lastname: Cheong
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Hirata
authors.forename: Abdur Rub Abdur
authors.initials: AR
authors.lastname: Rahman
authors.forename: Chwee Teck
authors.initials: CT
authors.lastname: Lim
mesh_terms: D019921:Biotinylation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D003470:Culture Media; D066246:ErbB Receptors / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D056656:Lab-On-A-Chip Devices; D008024:Ligands; D008027:Light; D008156:Luciferases / Q000378:metabolism; D049449:Luminescence; D044085:Microfluidics; D018613:Microscopy, Confocal; D032383:Myocytes, Cardiac / Q000166:cytology; D011485:Protein Binding; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D015203:Reproducibility of Results; D019809:Streptavidin / Q000737:chemistry; D013499:Surface Properties
chemical_list: D003470:Culture Media; D008024:Ligands; D011943:Receptors, Adrenergic, beta; D049452:Green Fluorescent Proteins; D019809:Streptavidin; D008156:Luciferases; D066246:ErbB Receptors
keywords: cell islands; cell surface receptors; gaussia luciferase; lens-free optics; microfluidics; receptor numbers