PicoQuant
Evangelos Sisamakis, Mathias Bayer, Matthias Patting, Marcelle Koenig, Marcus Sackrow, André Devaux, Uwe Ortmann, Felix Koberling, Rainer Erdmann
Workshop Room 5
Single molecule studies and – more specifically – single molecule FRET methodologies have become a standard tool for studying dynamic structural changes in proteins and nucleic acids. These types of measurements can reveal dynamic events on time scales covering several orders of magnitude from ~ns to several seconds. This allows studying e.g., chain dynamics, binding, folding, allosteric events, oligomerization, and aggregation. The power of these methodologies is highlighted by the study of Intrinsically Disordered Proteins (IDPs) whose biological relevance has been increasingly studied over the recent years.
In this remote workshop we will showcase how easy it is for new users to perform single molecule measurements on two model systems:
a) doubly labeled freely diffusing short oligonucleotides and
b) Cy5 molecules immobilized on the coverslip surface
Several online previews enable users to immediately judge sample and data quality. All correction parameters necessary to obtain FRET efficiency vs. stoichiometry histograms are automatically determined online, requiring no interaction from the user. The algorithm employs methodologies benchmarked by the scientific community.
Furthermore, we will show how the variable PSF feature can be used in smFRET and FCS measurements to fine-tune the observation window of freely diffusing biomolecules.
smFRET E-S-histogram
variable PSF
