Microscope for wide-field, single-molecule fluorescence imaging
This custom-designed instrument allows for epi- and through-the objective TIRF illuminated wide-field fluorescence excitation. Light is provided by 488, 532, 576 and635 nm CW lasers. Wavelengths can be switched using a acousto-optical tunable filter. Images are collected using a back-illuminated EMCCD. The instrument is set up for simultaneous detection of two of three different spectral channels. Optical filters are available for detection of EGFP/fluorescein/Alexa 488, Cy3/Alexa 555, mCherrry/Texas Red and Cy5/Alexa 647. The instrument was designed in particular for in vitro motility assays using kinesin and other motor proteins and imaging of single fluorophores in living cells.
Some essential parts of the laser setup are displayed below:
Contact person at LCVU: Erwin Peterman (erwinp@nat.vu.nl)