Demonstration of flat-top beam illumination in widefield multiphoton microscopy.

ABSTRACT: Multiphoton microscopy provides a suitable technique for imaging biological tissues with submicrometer resolution. Usually a Gaussian beam (GB) is used for illumination, leading to a reduced power efficiency in the multiphoton response and vignetting for a square-shaped imaging area. A flat-top beam (FTB) provides a uniform spatial intensity distribution that equalizes the probability of a multiphoton effect across the imaging area. We employ a customized widefield multiphoton microscope to compare the performance of a square-shaped FTB illumination with that based on using a GB, for both two-photon fluorescence (TPF) and second-harmonic generation (SHG) imaging. The variation in signal-to-noise ratio across TPF images of fluorescent dyes spans ∼5.6  dB for the GB and ∼1.2  dB for the FTB

SUBMITTER: Kabir M 

PROVIDER: S-EPMC7008505 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature