An easy and reliable way to perform single molecule FRET measurements.

Abstract

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 poster we show how easily these measurements can be performed with Luminosa single photon-counting confocal microscope and how all necessary correction parameters are automatically determined requiring no interaction from the user by employing methodologies benchmarked by the scientific community. We will also show how the variable PSF feature can be used in such measurements to fine-tune the observation window of freely diffusing biomolecules.