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SFB1032 Method Workshop on Fluorescence Spectroscopy and Microscopy
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Europe/Berlin
B 3.025 (LMU high tech Campus at Großhadern)
B 3.025
LMU high tech Campus at Großhadern
Butenandtstr. 5
Description
Fluorescence spectroscopy and microscopy offers very sensitive, non-contact approaches to investigate biological systems. Fluorescence benefits from extremely high sensitivity and specificity. In this workshop, we will give an introduction to four popular methods in fluorescence spectroscopy/microscopy: Fluorescence Correlation Spectroscopy (FCS), Fluorescence Lifetime Imaging Microscopy (FLIM), single-pair Förster Resonance Energy Transfer (spFRET) spectroscopy and Single Particle Tracking (SPT). In FCS, one analyzes the fluctuations in fluorescence intensity and can use it to monitor molecular diffusion, biomolecular interactions and kinetics. With FLIM, one uses the sensitivity of the fluorescence lifetime to investigate the nanoenvironment in the vicinity of the fluorophore. Images with optical resolution are collected with time-correlated single-photon counting to provide an image of the fluorescence lifetime. This can be used to monitor cellular function or biomolecular interactions. In spFRET, distances on the 2-10 nm scale can be measured, a distance perfect for investigating biomolecules. With the single molecule sensitivity of fluorescence, the conformation and dynamics of individual proteins and their interactions can be investigated. Using SPT, one can follow a single biomolecule and watch is as it performs its function.
In this mini-workshop, we will begin with lectures giving an introduction to the four different methods with examples demonstrating their application. In the afternoon, different computer exercises are offered to aid students in getting more familiar with the methods and how to analyze the data. Hence, attendees should bring a laptop with them. On the following morning, actual experiments will be demonstrated on the various microscopes so that the students can observe the methods in action.
