Fluorescence-based assays.
Fluorescence-based assays are widely used in high-throughput screening due to their high sensitivity, diverse selection of fluorophores, ease of operation, and various readout modes. As a result, fluorescence-based assays have been applied to monitor a broad range of activities in life-science research such as molecular dynamics and interactions, enzymatic activities, signal transduction, cell health, and distribution of molecules, organelles, or cells. This chapter describes two fluorescence-based techniques: total intensity measurement as an indication of cell viability, and fluorescence resonance energy transfer as an indication of protein folding and interactions, to illustrate in detail applications suitable for cell-based high-throughput screening in plate-reader and automated microscope-based formats, respectively.
An
W Frank
WF
Chemical Biology Platform, The Broad Institute of Harvard and MIT, Cambridge, MA, USA. fan@broad.harvard.edu
D001426:Bacterial Proteins / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects*; D008164:Luminescent Proteins / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D021381:Protein Transport
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D004364:Pharmaceutical Preparations; C054562:yellow fluorescent protein, Bacteria