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Bela T L Vogler1,2,3, Giovanni De Angelis4,5, Ziliang Zhao4,5,6

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MINFLUX super-resolution microscopy enables real-time Single Particle Tracking (SPT) of cellular dynamics. This study introduces a parameter optimization strategy for MINFLUX, successfully tracking fast lipid diffusion in membranes.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • MINFLUX (MFX) is a cutting-edge super-resolution microscopy technique.
  • It allows for detailed study of cellular structures and dynamics.
  • MINFLUX enables real-time Single Particle Tracking (SPT).

Purpose of the Study:

  • To propose a parameter optimization strategy for MINFLUX.
  • To provide an overview of critical MINFLUX parameters.
  • To demonstrate MFX-enabled SPT of fast-diffusing membrane lipids.

Main Methods:

  • Development of a parameter optimization strategy for MINFLUX.
  • Theoretical analysis of trackable diffusion rates.
  • Experimental application of MINFLUX SPT to membrane lipids.

Main Results:

  • A strategy for optimizing MINFLUX parameters is presented.
  • A theoretical limit for trackable diffusion rates was determined.
  • MINFLUX successfully tracked rapid lateral Brownian motion of lipids in membranes (D_MSD = 2.5 μm²/s).

Conclusions:

  • Optimized MINFLUX parameters are crucial for accurate measurements.
  • MINFLUX is capable of tracking fast molecular dynamics in cellular membranes.
  • This work advances the application of super-resolution microscopy in cell biology.