Fig. 1: Depth-variant deconvolution of thick image volumes acquired with a widefield microscope achieves sub-nuclear axial resolution for 3D visualization.
From: Depth-variant deconvolution applied to widefield microscopy for rapid large-volume tissue imaging
A Standard epifluorescent microscopy build with ×20 immersion lens (Carl Zeiss AG, NA 1.0, working distance of 6.4 mm) (top). Optical path for a refractive index-matched tissue mounted under a glass coverslip in contact with a ×20 lens through a water mount (bottom). B Extended display showing raw image of nuclei in a cleared mouse brain labeled with anti-Histone-H3-ATTO488 (green) acquired on widefield microscope before computational deconvolution. C Nuclei in cleared, transparent brain after deconvolution with depth-variant point spread functions calculated using Huygens Software (Scientific Volume Imaging) and visualized in the axial dimension with Imaris software. D Extended display showing resolution of fine microglial pseudopods labeled by CX3CR1-tdTomato (green) and nuclear staining with anti-Histone-H3-ATTO488 (magenta) captured using epifluorescent widefield microscopy with ×20 immersion lens (NA 1.0); E extended display of microglia as in (D) but captured using line-scanning confocal microscopy at a similar pixel resolution. F Maximum intensity projection of ~200 microns of 5XE4 leptomeninges attached to the brain surface that were stained for Aβ (red) to label CAA plaques along arterioles (smooth-muscle actin: green, CD31-positive vessels: gray), imaged using epifluorescent widefield microscopy with ×5 objective lens (NA 0.16, air), and deconvolved.
