Combining TIR and FRET: from fluorescence microscopy to a multi-well reader system

Herbert Schneckenburger; Petra Weber; Michael Wagner; Sandra Enderle; Julian Weghuber; Peter Lanzerstorfer

doi:10.1117/12.2526416

9 September 2019 Combining TIR and FRET: from fluorescence microscopy to a multi-well reader system

Herbert Schneckenburger, Petra Weber, Michael Wagner, Sandra Enderle, Julian Weghuber, Peter Lanzerstorfer

Proceedings Volume 11076, Advances in Microscopic Imaging II; 110761C (2019) https://doi.org/10.1117/12.2526416
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

Abstract

In view of a pharmacological test system with living cells we examined Förster Resonance Energy Transfer (FRET) between fluorescent proteins upon Total Internal Reflection (TIR) of a laser beam. We measured in particular HeLa cells expressing membrane associated EGFR-CFP and Grb2-YFP prior to and subsequent to stimulation with the epidermal growth factor EGF. In addition, HeLa cells expressing a membrane associated Green Fluorescent Protein (HeLa hFR-GPI-GFP) were incubated with the energy acceptor Nile Red for further validation. Measurements range from fluorescence microscopy to Fluorescence Lifetime Imaging (FLIM) of a larger number of samples in a TIR reader.

Citation Download Citation

Herbert Schneckenburger, Petra Weber, Michael Wagner, Sandra Enderle, Julian Weghuber, and Peter Lanzerstorfer "Combining TIR and FRET: from fluorescence microscopy to a multi-well reader system", Proc. SPIE 11076, Advances in Microscopic Imaging II, 110761C (9 September 2019); https://doi.org/10.1117/12.2526416

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