Axial resolution and contrast enhancement in digital scanned light-sheet microscopy via stimulated emission depletion

Abstract

Integration of stimulated emission depletion (STED) with digital scanned light-sheet microscopy (DSLM) using a Bessel beam achieves fast, high-resolution and large field of view (FOV) imaging. Here, we report on a combination method of DSLM and STED by using a Bessel beam for excitation and a hollow Gaussian beam for depletion. Owing to the advantages of confined axial distribution around the focal plane for the hollow Gaussian depletion beam, those fluorescence signals from the periphery of the Bessel excitation pattern are depleted by stimulated emission. A super-resolved light sheet with suppressed side lobes is achieved. The numerical simulation results show that the use of the Bessel beam for the excitation and the hollow Gaussian beam for the STED results in a DSLM system with enhanced performance of the axial resolution and image contrast over a moderately large FOV.

Enhancement of image quality in planar Airy light-sheet microscopy via subtraction method

“…The polarizing beam splitters (PBS1 and PBS2) are transparent for the x-polarized part and reflective for the y-polarized part. For creating a hollow light-sheet, a semicircular half-wavelength phase plate (SPP) with a π-phase retardation between the two equal parts of the incoming wavefront is inserted in the transparent x-polarized beam where the divided line of the phase delay is set parallel to the x-polarization [21,22]. The phase retardation introduced by SPP is…”