Imaging of molecular mobility by spatial Fourier transform fluorescence recovery after photobleaching (FT-FRAP) with structured illumination

Garth J. Simpson

doi:10.1117/12.3005854

13 March 2024 Imaging of molecular mobility by spatial Fourier transform fluorescence recovery after photobleaching (FT-FRAP) with structured illumination

Garth J. Simpson

Author Affiliations +

Proceedings Volume PC12855, Advanced Chemical Microscopy for Life Science and Translational Medicine 2024; PC128550P (2024) https://doi.org/10.1117/12.3005854
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

FT-FRAP with structured illumination is demonstrated for recovery of 2D molecular diffusion maps within heterogeneous systems. Distribution of the bleach power in comb illumination patterns maintained high integrated power, but avoids “hot spots” from local heating typical in point-bleach multi-photon excitation. Spatial Fourier transform approaches support recovery of diffusion coefficient imaging, in which 2D maps of spatially varying diffusion coefficients are recovered within a given field of view. Proof-of-concept simulations with known ground-truth behaviors enable algorithm assessment. Experiments are described to characterize molecular and macromolecular mobility within biological tissues and tissue-mimetic materials. Additional applications include mapping of small molecule diffusion within model heterogeneous pharmaceutical formulations to understand bioavailability and stability.

Conference Presentation

Citation Download Citation

Garth J. Simpson "Imaging of molecular mobility by spatial Fourier transform fluorescence recovery after photobleaching (FT-FRAP) with structured illumination", Proc. SPIE PC12855, Advanced Chemical Microscopy for Life Science and Translational Medicine 2024, PC128550P (13 March 2024); https://doi.org/10.1117/12.3005854

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
PRESENTATION

WATCH
PRESENTATION SAVE TO MY LIBRARY

GET CITATION

KEYWORDS

Light sources and illumination

Biological imaging

Fourier transforms

Diffusion

Fluorescence

Fluorescence imaging

Photobleaching

Show All Keywords

Keywords/Phrases

Search In:

Publication Years