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Single Molecule FRET

Multicolor Confocal

This home-built device is based on an olympus body. The microscope is used for in solution FRET measurements using two picosecond pulsed lasers (483 nm and 635 nm). Molecules (or any labeled structure) diffusing through the focal volume are recorded in a time resolved manner and measurements include anisotropy information. This is achieved by using four single photon detectors, two for each FRET channel. The main applications are FCS and single-molecule fluorescence.

Relevant publications:

Hellenkamp, B., Schmid, S., Doroshenko, O., Opanasyuk, O., Kühnemuth, R., Rezaei Adariani, S., … Hugel, T. (2018). Precision and accuracy of single-molecule FRET measurements-a multi-laboratory benchmark study. Nature methods, 15(9), 669–676. doi:10.1038/s41592-018-0085-0

Vandenberk, N., Barth, A., Borrenberghs, D., Hofkens, J., Hendrix, J.,  (2018). Evaluation of Blue and Far-Red Dye Pairs in Single-Molecule Förster Resonance Energy Transfer Experiments,

The Journal of Physical Chemistry, 8 122 (15), 4249-4266. doi:10.1021/acs.jpcb.8b00108

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