Modal focal adaptive optics for Bessel-focus two-photon fluorescence microscopy

Kim, Hyeonggeon; Natan, Ryan; Chen, Wei; Winans, Amy M.; Fan, Jiang Lan; Isacoff, Ehud; Ji, Na

doi:10.1364/OE.541033

Title: Modal focal adaptive optics for Bessel-focus two-photon fluorescence microscopy

Journal Article · 07 January 2025 · Optics Express

DOI: https://doi.org/10.1364/OE.541033 · OSTI ID:2497352

; Natan, Ryan; Chen, Wei; Winans, Amy M.; Fan, Jiang Lan; Isacoff, Ehud;

Adaptive optics (AO) improves the spatial resolution of microscopy by correcting optical aberrations. While its application has been well established in microscopy modalities utilizing a circular pupil, its adaptation to systems with non-circular pupils, such as Bessel-focus two-photon fluorescence microscopy (2PFM) with an annular pupil, remains relatively uncharted. Herein, we present a modal focal AO (MFAO) method for Bessel-focus 2PFM. Measuring and correcting aberration using a spatial light modulator placed in conjugation with the focal plane of the microscope objective, MFAO employs Zernike annular polynomials — a first in AO implementation — to achieve performance on par with a previous zonal AO method, but with a notably simplified optical configuration. We validated the performance of MFAO in correcting artificial and sample-induced aberrations, as well as in in vivo imaging of zebrafish larvae and mouse brains. By expanding the application of modal AO to annular pupils as well as aberration measurement and correction to a wavefront modulator at the objective focal plane, MFAO represents a notable advancement in the implementation of AO in microscopy.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: SC0021986

OSTI ID:: 2497352

Journal Information:: Optics Express, Journal Name: Optics Express Journal Issue: 1 Vol. 33; ISSN 1094-4087; ISSN OPEXFF

Publisher:: Optical Society of AmericaCopyright Statement

Country of Publication:: United States

Language:: English

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