Diffusion mapping by fourier-transform FRAP with multi-photon excited structured illumination

Garth J. Simpson

doi:10.1117/12.2610488

7 March 2022 Diffusion mapping by fourier-transform FRAP with multi-photon excited structured illumination

Garth J. Simpson

Proceedings Volume PC11965, Multiphoton Microscopy in the Biomedical Sciences XXII; PC119650R (2022) https://doi.org/10.1117/12.2610488
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

We demonstrate an approach for producing spatially resolved maps of macromolecular mobility compatible with multi-photon excitation in fluorescence recovery after photobleaching (FRAP). Periodically structured illumination during photobleaching allows distribution of the photobleach power over the entire field of view to reduce local heating effects, but recovers highly localized peaks in the spatial Fourier transform domain. Recentering the FT peaks and inverse Fourier transformation yields images scaling with the amplitude of the photobleach modulation at each spatial location. From the temporal decays, up to 250,000 independent diffusion coefficients can be recovered for each position in the field of view.

Conference Presentation

Citation Download Citation

Garth J. Simpson "Diffusion mapping by fourier-transform FRAP with multi-photon excited structured illumination", Proc. SPIE PC11965, Multiphoton Microscopy in the Biomedical Sciences XXII, PC119650R (7 March 2022); https://doi.org/10.1117/12.2610488

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