A spherical aberration-free microscopy system for live brain imaging.

A spherical aberration-free microscopy system for live brain imaging. Biochem Biophys Res Commun. 2018 Apr 09;: Authors: Ue Y, Monai H, Higuchi K, Nishiwaki D, Tajima T, Okazaki K, Hama H, Hirase H, Miyawaki A Abstract The high-resolution in vivo imaging of mouse brain for quantitative analysis of fine structures, such as dendritic spines, requires objectives with high numerical apertures (NAs) and long working distances (WDs). However, this imaging approach is often hampered by spherical aberration (SA) that results from the mismatch of refractive indices in the optical path and becomes more severe with increasing depth of target from the brain surface. Whereas a revolving objective correction collar has been designed to compensate SA, its adjustment requires manual operation and is inevitably accompanied by considerable focal shift, making it difficult to acquire the best image of a given fluorescent object. To solve the problems, we have created an objective-attached device and formulated a fast iterative algorithm for the realization of an automatic SA compensation system. The device coordinates the collar rotation and the Z-position of an objective, enabling correction collar adjustment while stably focusing on a target. The algorithm provides the best adjustment on the basis of the calculated contrast of acquired images. Together, they enable the system to compensate SA at a given depth. As proof of concept, we applied the SA c...
Source: Biochemical and Biophysical Research communications - Category: Biochemistry Authors: Tags: Biochem Biophys Res Commun Source Type: research