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Single Molecule Microscope - TIRF

Description

for single-molecule FRET and super-resolution microscopy

Our single-molecule microscopy (SMM) setup is a home-built total internal reflection fluorescence (TIRF) microscope with 4 (alternating) laser excitation channels and three emission channels. It can therefore be used for single-molecule Förster Resonance Energy Transfer (FRET) spectroscopy, as well as super-resolution microscopy techniques such as photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM).

 Principle of TIRF

In TIRF microscopy the laser light is focused into the rim of the backfocal plane of the microscope objective such as that the laser light is totally reflected at the glass/ water interface. An evanescent field is created in which intensity decays within a few hundred nanometre. Thereby, only a small volume above the coverslide is illuminated.