Single Molecule FRET

This lecture covers the fundamentals of Biophotonics, focusing on Single Molecule FRET (Förster Resonance Energy Transfer) with Total Internal Reflection Microscopy. It explains the non-radiative energy transfer between molecules, the conditions required for energy transfer, and the selection of appropriate fluorophores. The lecture also discusses the applications of FRET in studying protein structure, protein complexes, receptor interactions, and nucleic acids. It delves into the labeling of molecules for FRET experiments, the advantages of single molecule FRET, and the techniques for surface immobilization. Additionally, it explores the principles of Total Internal Reflection Fluorescence (TIRF) microscopy and its applications in imaging cells. The lecture concludes with a discussion on TIRF instrumentation configurations and practical examples of TIRF microscopy.