1. A brief introduction to single-molecule fluorescence methods
- Author
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van den Wildenberg, Siet M.J.L., Prevo, Bram, Peterman, Erwin J.G., Peterman, Erwin J.G., Physics of Living Systems, and LaserLaB - Molecular Biophysics
- Subjects
0301 basic medicine ,Physics ,Microscopy ,Total internal reflection fluorescence microscope ,Fluorophore ,TIRF ,010402 general chemistry ,Single-molecule experiment ,01 natural sciences ,Fluorescence ,0104 chemical sciences ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,Förster resonance energy transfer ,Confocal fluorescence ,chemistry ,Wide-field epi-fluorescence ,Fluorescence microscope ,Biological system ,Fluorescence anisotropy - Abstract
One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.
- Published
- 2018