"MINFLUX for Dynamic Structural Cell Biology"

Jonas Ries
University of Vienna
 

Abstract: MINFLUX can localize single fluorophores with unprecedented precision by targeted detection with a scanned, patterned beam. In combination with switchable fluorophores, this allows for super-resolution imaging with single nanometer resolution, and has been extended to 3D and multiple colors. As MINFLUX uses the photon budget of a single fluorophore very efficiently, it is also a very promising technique for single-fluorophore tracking, improving speed, precision and track length by one order of magnitude compared to camera-based tracking.

Here, I will introduce the principle of MINFLUX and its opportunities and limitations for dynamic cellular imaging. I will then discuss how we used MINFLUX to track the stepping motion of the motor protein kinesin-1 as it walks on microtubules in living cells and will show first results on dual-color co-tracking to directly monitor conformational changes of proteins. I will discuss technical aspects of how we characterize the stability of our setup and will end by presenting a new approach to MINLFLUX that we will develop into an open-source MINFLUX microscope.
 

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