Streamlined Single-Molecule FRET and FCS Measurements for Structural Biology and Phase Separation Studies
Quantitative single-molecule and time-resolved fluorescence techniques offer new insights into many samples from various research areas such as dynamic structural biology, cellular mechanisms driven by phase separation, and virology. Using examples from these areas, Dr. Maria Loidolt-Krüger shows how single-molecule FRET (smFRET), fluorescence (cross-)correlation spectroscopy, and time-resolved anisotropy can be combined to get a comprehensive picture of the sample under investigation.
In order to get reliable, quantitative results, many factors like laser power, or spectral bleed-through need to be considered. These will be discussed, together with ways to increase the accuracy and reproducibility of experiments, with a focus on smFRET studies. Ultimately, findings from in vitro studies need to be linked to studies in cells and tissues. Maria showcases how this can be achieved by taking FCS into cells, and by going from smFRET to FRET imaging, and from time-resolved anisotropy to anisotropy imaging.
The presentation is available on demand on our website.