Publication

Dual-color Total Internal Reflection Fluorescence Fluctuation Spectroscopy

Theo Lasser, Marcel Leutenegger
2006
Talks

Abstract

We developed a dual-color total internal reflection fluorescence (TIRF) system for single molecule imaging and fluorescence fluctuation spectroscopy (FFS). For a performance analysis, we measured a synthetic binding assay with dual-color global fluorescence correlation spectroscopy (2CG-FCS) and dual-color fluorescence intensity distribution analysis (2D-FIDA). We show that dual-color TIR-FFS is a suitable method for measuring coincidence assays at the surface. In particular, our setup has a very high fluorescence collection efficiency resulting in a two- to three-fold increased molecular brightness. Using a customized workbench setup, we improved the setup alignment, the sample handling and the system versatility.

Official source

https://infoscience.epfl.ch/entities/publication/43b6e9b5-cc6b-4210-90bd-3264def1300c

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Theo Lasser, Marcel Leutenegger
2006
Talks

Abstract

Official source

https://infoscience.epfl.ch/entities/publication/43b6e9b5-cc6b-4210-90bd-3264def1300c

About this result

Related concepts (30)

Fluorescence spectroscopy

Fluorescence spectroscopy (also known as fluorimetry or spectrofluorometry) is a type of electromagnetic spectroscopy that analyzes fluorescence from a sample. It involves using a beam of light, usually ultraviolet light, that excites the electrons in molecules of certain compounds and causes them to emit light; typically, but not necessarily, visible light. A complementary technique is absorption spectroscopy. In the special case of single molecule fluorescence spectroscopy, intensity fluctuations from the emitted light are measured from either single fluorophores, or pairs of fluorophores.

Super-resolution microscopy

Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that use the Pendry Superlens and near field scanning optical microscopy) or on the far-field.

Förster resonance energy transfer

Förster resonance energy transfer (FRET), fluorescence resonance energy transfer, resonance energy transfer (RET) or electronic energy transfer (EET) is a mechanism describing energy transfer between two light-sensitive molecules (chromophores). A donor chromophore, initially in its electronic excited state, may transfer energy to an acceptor chromophore through nonradiative dipole–dipole coupling. The efficiency of this energy transfer is inversely proportional to the sixth power of the distance between donor and acceptor, making FRET extremely sensitive to small changes in distance.

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