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Affordable optical clearing and immunolabelling in mouse brain slices
Traditional histological analysis is conducted on thin tissue sections, limiting the data capture from large tissue volumes to 2D profiles, and requiring stereological methods for 3D assessment. Recent advances in microscopical and tissue clearing methods have facilitated 3D reconstructions of tissu...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10543858/ https://www.ncbi.nlm.nih.gov/pubmed/37777793 http://dx.doi.org/10.1186/s13104-023-06511-y |
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author | Muza, Phillip M. Pérez, Marta Noy, Suzanna Kurosawa, Miyu Katsouri, Loukia Tybulewicz, Victor L. J. Fisher, Elizabeth M.C. West, Steven J. |
author_facet | Muza, Phillip M. Pérez, Marta Noy, Suzanna Kurosawa, Miyu Katsouri, Loukia Tybulewicz, Victor L. J. Fisher, Elizabeth M.C. West, Steven J. |
author_sort | Muza, Phillip M. |
collection | PubMed |
description | Traditional histological analysis is conducted on thin tissue sections, limiting the data capture from large tissue volumes to 2D profiles, and requiring stereological methods for 3D assessment. Recent advances in microscopical and tissue clearing methods have facilitated 3D reconstructions of tissue structure. However, staining of large tissue blocks remains a challenge, often requiring specialised and expensive equipment to clear and immunolabel tissue. Here, we present the Affordable Brain Slice Optical Clearing (ABSOC) method: a modified iDISCO protocol which enables clearing and immunolabeling of mouse brain slices up to 1 mm thick using inexpensive reagents and equipment, with no intensive expert training required. We illustrate the use of ABSOC in 1 mm C57BL/6J mouse coronal brain slices sectioned through the dorsal hippocampus and immunolabelled with an anti-calretinin antibody. The ABSOC method can be readily used for histological studies of mouse brain in order to move from the use of very thin tissue sections to large volumes of tissue – giving more representative analysis of biological samples, without the need for sampling of small regions only. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-023-06511-y. |
format | Online Article Text |
id | pubmed-10543858 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-105438582023-10-03 Affordable optical clearing and immunolabelling in mouse brain slices Muza, Phillip M. Pérez, Marta Noy, Suzanna Kurosawa, Miyu Katsouri, Loukia Tybulewicz, Victor L. J. Fisher, Elizabeth M.C. West, Steven J. BMC Res Notes Research Note Traditional histological analysis is conducted on thin tissue sections, limiting the data capture from large tissue volumes to 2D profiles, and requiring stereological methods for 3D assessment. Recent advances in microscopical and tissue clearing methods have facilitated 3D reconstructions of tissue structure. However, staining of large tissue blocks remains a challenge, often requiring specialised and expensive equipment to clear and immunolabel tissue. Here, we present the Affordable Brain Slice Optical Clearing (ABSOC) method: a modified iDISCO protocol which enables clearing and immunolabeling of mouse brain slices up to 1 mm thick using inexpensive reagents and equipment, with no intensive expert training required. We illustrate the use of ABSOC in 1 mm C57BL/6J mouse coronal brain slices sectioned through the dorsal hippocampus and immunolabelled with an anti-calretinin antibody. The ABSOC method can be readily used for histological studies of mouse brain in order to move from the use of very thin tissue sections to large volumes of tissue – giving more representative analysis of biological samples, without the need for sampling of small regions only. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-023-06511-y. BioMed Central 2023-09-30 /pmc/articles/PMC10543858/ /pubmed/37777793 http://dx.doi.org/10.1186/s13104-023-06511-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Note Muza, Phillip M. Pérez, Marta Noy, Suzanna Kurosawa, Miyu Katsouri, Loukia Tybulewicz, Victor L. J. Fisher, Elizabeth M.C. West, Steven J. Affordable optical clearing and immunolabelling in mouse brain slices |
title | Affordable optical clearing and immunolabelling in mouse brain slices |
title_full | Affordable optical clearing and immunolabelling in mouse brain slices |
title_fullStr | Affordable optical clearing and immunolabelling in mouse brain slices |
title_full_unstemmed | Affordable optical clearing and immunolabelling in mouse brain slices |
title_short | Affordable optical clearing and immunolabelling in mouse brain slices |
title_sort | affordable optical clearing and immunolabelling in mouse brain slices |
topic | Research Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10543858/ https://www.ncbi.nlm.nih.gov/pubmed/37777793 http://dx.doi.org/10.1186/s13104-023-06511-y |
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