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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Optical Society of America
2021
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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. |
format | Online Article Text |
id | pubmed-8713664 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Optical Society of America |
record_format | MEDLINE/PubMed |
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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