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, Loidolt-Krüger will show 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. Loidolt-Krüger will showcase 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.

About the presenter
Maria Loidolt-Krüger, Ph.D., studied biophysics at the Technical University Kaiserslautern. During an exchange year at the University of Southern Denmark in Odense, she dived into molecular disease diagnostics. She then worked on her diploma and doctorate thesis in the group of Prof. Stefan W. Hell at the Max Planck Institute for Biophysical Chemistry in Göttingen.

During her thesis, she developed multicolor STED microscopy and the corresponding live cell staining protocol. While pursuing her doctorate, Loidolt-Krüger studied the feasibility of performing STED microscopy of FRET pairs. In 2018 she joined PicoQuant as an application specialist for time-resolved fluorescence microscopes. In 2022, she changed roles and is now scientific content creator at PicoQuant, responsible for all product lines for life and materials science.