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Simultaneous scattering compensation at multiple points in multi-photon microscopy

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...

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Autores principales: May, Molly A., Kummer, Kai K., Edenhofer, Marie-Luise, Choconta, Jeiny Luna, Kress, Michaela, Ritsch-Marte, Monika, Jesacher, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Optical Society of America 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8713664/
https://www.ncbi.nlm.nih.gov/pubmed/35003840
http://dx.doi.org/10.1364/BOE.441604
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author May, Molly A.
Kummer, Kai K.
Edenhofer, Marie-Luise
Choconta, Jeiny Luna
Kress, Michaela
Ritsch-Marte, Monika
Jesacher, Alexander
author_facet May, Molly A.
Kummer, Kai K.
Edenhofer, Marie-Luise
Choconta, Jeiny Luna
Kress, Michaela
Ritsch-Marte, Monika
Jesacher, Alexander
author_sort May, Molly A.
collection PubMed
description 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.
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spelling pubmed-87136642022-01-07 Simultaneous scattering compensation at multiple points in multi-photon microscopy May, Molly A. Kummer, Kai K. Edenhofer, Marie-Luise Choconta, Jeiny Luna Kress, Michaela Ritsch-Marte, Monika Jesacher, Alexander Biomed Opt Express Article 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. Optical Society of America 2021-11-09 /pmc/articles/PMC8713664/ /pubmed/35003840 http://dx.doi.org/10.1364/BOE.441604 Text en Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/
spellingShingle Article
May, Molly A.
Kummer, Kai K.
Edenhofer, Marie-Luise
Choconta, Jeiny Luna
Kress, Michaela
Ritsch-Marte, Monika
Jesacher, Alexander
Simultaneous scattering compensation at multiple points in multi-photon microscopy
title Simultaneous scattering compensation at multiple points in multi-photon microscopy
title_full Simultaneous scattering compensation at multiple points in multi-photon microscopy
title_fullStr Simultaneous scattering compensation at multiple points in multi-photon microscopy
title_full_unstemmed Simultaneous scattering compensation at multiple points in multi-photon microscopy
title_short Simultaneous scattering compensation at multiple points in multi-photon microscopy
title_sort simultaneous scattering compensation at multiple points in multi-photon microscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8713664/
https://www.ncbi.nlm.nih.gov/pubmed/35003840
http://dx.doi.org/10.1364/BOE.441604
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