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Q: Fachverband Quantenoptik und Photonik

Q 41: Ultrashort Laser Pulses: Applications III

Q 41.10: Vortrag

Mittwoch, 10. März 2010, 18:45–19:00, F 342

Colocalization analyses by two-color STED microscopy — •Johanna Bückers, Daniel Neumann, Stefan Jakobs, Lars Kastrup, and Stefan W. Hell — Max Planck Institute for Biophysical Chemistry, Dept. of NanoBiophotonics, Am Fassberg 11, 37077 Göttingen, Germany

Far-field fluorescence microscopy is among the most common methods in the biosciences today. One of its few drawbacks − the relatively poor spatial resolution − has been overcome since the invention of stimulated emission depletion (STED) microscopy and other nanoscopy concepts which allow imaging on the nanoscale.

Early STED microscopes were based on rather complex setups and high-maintenance laser systems, but the advent of supercontinuum laser sources enable the implementation of compact and tunable STED microscopes. Even further, due to the broad spectrum of available wavelengths two-color imaging can be realized comfortably, such that the interplay of e. g. different proteins in biological samples can be studied. For such colocalization analyses special care has to be taken regarding the cross-talk between differently labeled structures, and one has to ensure that there is no shift of the two images during the recording. Therefore, we developed an approach to quasi-simultaneous two-color STED microscopy, which allows cross-talk correction by means of linear unmixing, based on a pulse-interleaved acquisition scheme. This approach is applied for several biological tasks, e. g. to reveal the different degrees of colocalization of proteins in the mitochondrial membrane.