Journal of Innovative Optical Health Sciences, 2014, 7 (5): 1450025, Published Online: Jan. 10, 2019  

An automated multiwell plate reading flim microscope for live cell autofluorescence lifetime assays

Author Affiliations
1 Photonics Group, Department of Physics, Imperial College London South Kensington Campus, London, SW7 2AZ, UK
2 Institute of Chemical Biology, Department of Chemistry Imperial College London, South Kensington Campus London, SW7 2AZ, UK
3 Department of Surgery and Cancer, Imperial College London Du Cane Road, London, W12 0NN, UK
4 National Heart & Lung Institute Imperial Centre for Experimental & Translational Medicine Du Cane Road, London, W12 0NN, UK
5 Centre for Histopathology, Imperial College London Du Cane Road, London, W12 0NN, UK
Abstract
Fluorescence lifetime imaging (FLIM) is increasingly used to read out cellular autofluorescence originating from the coenzyme NADH in the context of investigating cell metabolic state. We present here an automated multiwell plate reading FLIM microscope optimized for UV illumination with the goal of extending high content fluorescence lifetime assays to readouts of metabolism. We demonstrate its application to automated cellular autofluorescence lifetime imaging and discuss the key practical issues associated with its implementation. In particular, we illustrate its capability to read out the NADH-lifetime response of cells to metabolic modulators, thereby illustrating the potential of the instrument for cytotoxicity studies, assays for drug discovery and stratified medicine.

Douglas J. Kelly, Sean C. Warren, Sunil Kumar, Joao L. Lagarto, Benjamin T. Dyer, Anca Margineanu, Eric W.-F. Lam, Chris Dunsby, Paul M. W. French. An automated multiwell plate reading flim microscope for live cell autofluorescence lifetime assays[J]. Journal of Innovative Optical Health Sciences, 2014, 7(5): 1450025.

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