Active image optimization for lattice light sheet microscopy in thick samples

Maxime Malivert, Fabrice Harms, Cynthia Veilly, Jerome Legrand, Ziqiang Li, Emmanuelle Bayer, Daniel Choquet, Mathieu Ducros
Biomed. Opt. Express. 2022-11-07; 13(12): 6211
DOI: 10.1364/BOE.471757

Lattice light-sheet microscopy (LLSM) is a very efficient technique for high resolution 3D imaging of dynamic phenomena in living biological samples. However, LLSM imaging remains limited in depth due to optical aberrations caused by sample-based refractive index mismatch. Here, we propose a simple and low-cost active image optimization (AIO) method to recover high resolution imaging inside thick biological samples. AIO is based on (1) a light-sheet autofocus step (AF) followed by (2) an adaptive optics image-based optimization. We determine the optimum AIO parameters to provide a fast, precise and robust aberration correction on biological samples. Finally, we demonstrate the performances of our approach on sub-micrometric structures in brain slices and plant roots.

Auteurs Bordeaux Neurocampus