Widefield FRET & TIRF Microscope
The Zeiss Axiovert 200M is configured to be capable of widefield Fluorescence Resonance Energy Transfer (FRET) microscopy and Total Internal Reflection Fluorescence (TIRF) microscopy. The microscope is equipped with a cooled Photometrics 512 Evolve EMCCD camera with high sensitivity for quantitative low-light fluorescence imaging.
FRET microscopy allows the determination of the approach between two molecules within several nanometers, a distance sufficiently close for molecular interactions to occur. For the FRET microscopy, the imaging and system control is realized by using the software MetaFluor. The filter sets installed in the microscope allow imaging a number of FRET pairs, such as ECFP-YPet.
TIRF microscopy can be used to observe the fluorescence of a single molecule or a thin region of a specimen, usually less than 200 nanometers thick. Currently three lasers (the wavelengths are 405 nm, 473 nm and 561 nm) are available for the TIRF imaging.