Estimating the point-spread-function using 1 µm diameter microspheres for image restoration in biomedical multiphoton microscopy

Ségolène Martin; Julien Ajdenbaum; Emilie Chouzenoux; Laetitia Magnol; Véronique Blanquet; Jean-Christophe Pesquet; Claire Lefort

doi:10.1117/12.3016624

18 June 2024 Estimating the point-spread-function using 1 μm diameter microspheres for image restoration in biomedical multiphoton microscopy

Ségolène Martin, Julien Ajdenbaum, Emilie Chouzenoux, Laetitia Magnol, Véronique Blanquet, Jean-Christophe Pesquet, Claire Lefort

Author Affiliations +

Proceedings Volume 12998, Optics, Photonics, and Digital Technologies for Imaging Applications VIII; 129980L (2024) https://doi.org/10.1117/12.3016624
Event: SPIE Photonics Europe, 2024, Strasbourg, France

Abstract

Multiphoton microscopy (MPM) is an advanced imaging technique used in biological and biomedical research to visualize and study living tissues and cells with high resolution. It is particularly well-suited for deep tissue imaging, as it minimizes photodamage and provides improved penetration compared to traditional microscopy techniques like confocal microscopy. The Point Spread Function (PSF) plays a crucial role in multiphoton microscopy and describes how a point source of light is imaged as a spatial distribution in the microscope. Understanding the 3D PSF is essential for deconvolution and other post-processing techniques used to reconstruct 3D images from a stack of 2D images. The imaging of fluorescent beads used as point sources is a solution to evaluate the PSF. Usually, this strategy involves a bead of dimensions smaller than resolutions, typically with diameter of about less than 200 nm. However, in this setting, it is often complex to detect correctly the beads and thus to estimate accurately the PSF. We develop a computational solution for the PSF estimation based on the imaging of micro objects bigger than the resolution limit. We use fluorescent microspheres with a diameter of 1 μm and estimate the PSF from the deformations observed in the image of these microspheres. A deconvolution strategy illustrates the performance of our method, where we successfully restore an unsliced whole striated skeletal muscle utilizing the PSF estimated with 1 μm diameter beads.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Ségolène Martin, Julien Ajdenbaum, Emilie Chouzenoux, Laetitia Magnol, Véronique Blanquet, Jean-Christophe Pesquet, and Claire Lefort "Estimating the point-spread-function using 1 μm diameter microspheres for image restoration in biomedical multiphoton microscopy", Proc. SPIE 12998, Optics, Photonics, and Digital Technologies for Imaging Applications VIII, 129980L (18 June 2024); https://doi.org/10.1117/12.3016624

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