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Simultaneous scattering compensation at multiple points in multi-photon microscopy

Authors :
Michaela Kress
Marie-Luise Edenhofer
Molly A. May
Jeiny Luna Choconta
Kai K. Kummer
Monika Ritsch-Marte
Alexander Jesacher
Source :
Biomedical Optics Express
Publication Year :
2021
Publisher :
Optical Society of America, 2021.

Abstract

The two-photon fluorescence imaging depth has been significantly improved in recent years by compensating for tissue scattering with wavefront correction. However, in most approaches the wavefront corrections are valid only over a small sample region on the order of 1 to 10 µm. In samples where most scattering structures are confined to a single plane, sample conjugate correction geometries can increase the observable field to a few tens of µm. Here, we apply a recently introduced fast converging scheme for sensor-less scattering correction termed “Dynamic Adaptive Scattering compensation Holography” (DASH) in a sample conjugate configuration with a high pixel count nematic liquid crystal spatial light modulator (LC-SLM). Using a large SLM allows us to simultaneously correct for scattering at multiple field points, which can be distributed over the entire field of view provided by the objective lens. Despite the comparably slow refresh time of LC-SLMs, we achieve correction times on the order of 10 s per field point, which we show is sufficiently fast to counteract scattering at multiple sites in living mouse hippocampal tissue slices.

Details

Language :
English
ISSN :
21567085
Volume :
12
Issue :
12
Database :
OpenAIRE
Journal :
Biomedical Optics Express
Accession number :
edsair.doi.dedup.....a68811ceb68861f31534f27683b0e36a