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1Photonics Group, Department of Physics,Imperial College London, 2Institute for Chemical Biology, Department of Chemistry,Imperial College London, 3MRC Clinical Sciences Centre,Hammersmith Hospital, 4Chemical Biology Section, Department of Chemistry,Imperial College London, 5Retroscreen Virology Ltd, 6Pfizer Global Research and Development,Pfizer Limited, Sandwich, Kent, UK, 7Centre for Histopathology,Imperial College London
Chapters
- 00:05Title
- 01:03Open Fluorescence Lifetime Imaging (FLIM)–High Content Analysis (HCA) Plugin Setup
- 01:51Instrument Response Function (IRF) Image and Reference Dye Data Acquisition
- 04:32Multiwell Plate Sample FLIM Data Acquisition
- 06:29Results: Representative FLIM Analyses in Cos and HeLa Cells
- 08:05Conclusion
We present an open source high content analysis (HCA) instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using Förster resonance energy transfer (FRET) based readouts. Data acquisition for this openFLIM-HCA instrument is controlled by software written in µManager and data analysis is undertaken in FLIMfit.
Tags
Keywords:
Open SourceHigh Content AnalysisAutomatedFluorescence Lifetime Imaging MicroscopyFLIMMulti-well PlateGlobal AnalysisFRETTime-gatedLive Cell ImagingProtein InteractionsCell SignalingMicromanagerOpenFLIM-HCA PluginDelay Box CalibrationInstrument Response FunctionSpinning Disk UnitFast Delay Box
