Precisely calibrated and spatially informed illumination for conventional fluorescence and improved PALM imaging applications.

ABSTRACT: Spatial light modulation using cost efficient digital micromirror devices (DMD) is finding broad applications in fluorescence microscopy due to the reduction of phototoxicity and bleaching and the ability to manipulate proteins in optogenetic experiments. However, precise illumination by DMDs and their application to single-molecule localization microscopy (SMLM) remained a challenge because of non-linear distortions between the DMD and camera coordinate systems caused by optical components in the excitation and emission path. Here we develop a fast and easy to implement calibration procedure that determines these distortions and matches the DMD and camera coordinate system with a precision below the optical diffraction limit. As a result, a region from a fluorescence image can be selected

SUBMITTER: Mancebo A 

PROVIDER: S-EPMC8609923 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature