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A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping

The mammalian brain, with its complexity and intricacy, poses significant challenges for researchers aiming to understand its inner workings. Optical multilayer interference tomography (OMLIT) is a novel, promising imaging technique that enables the mapping and reconstruction of mesoscale all-cell b...

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Autores principales: Wang, Tianyi, Shi, Peiyao, Luo, Dingsan, Guo, Jun, Liu, Hui, Yuan, Jinyun, Jin, Haiqun, Wu, Xiaolong, Zhang, Yueyi, Xiong, Zhiwei, Zhu, Jinlong, Zhou, Renjie, Zhang, Ruobing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10216590/
https://www.ncbi.nlm.nih.gov/pubmed/37239183
http://dx.doi.org/10.3390/brainsci13050711
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author Wang, Tianyi
Shi, Peiyao
Luo, Dingsan
Guo, Jun
Liu, Hui
Yuan, Jinyun
Jin, Haiqun
Wu, Xiaolong
Zhang, Yueyi
Xiong, Zhiwei
Zhu, Jinlong
Zhou, Renjie
Zhang, Ruobing
author_facet Wang, Tianyi
Shi, Peiyao
Luo, Dingsan
Guo, Jun
Liu, Hui
Yuan, Jinyun
Jin, Haiqun
Wu, Xiaolong
Zhang, Yueyi
Xiong, Zhiwei
Zhu, Jinlong
Zhou, Renjie
Zhang, Ruobing
author_sort Wang, Tianyi
collection PubMed
description The mammalian brain, with its complexity and intricacy, poses significant challenges for researchers aiming to understand its inner workings. Optical multilayer interference tomography (OMLIT) is a novel, promising imaging technique that enables the mapping and reconstruction of mesoscale all-cell brain atlases and is seamlessly compatible with tape-based serial scanning electron microscopy (SEM) for microscale mapping in the same tissue. However, currently, OMLIT suffers from imperfect coatings, leading to background noise and image contamination. In this study, we introduced a new imaging configuration using carbon spraying to eliminate the tape-coating step, resulting in reduced noise and enhanced imaging quality. We demonstrated the improved imaging quality and validated its applicability through a correlative light–electron imaging workflow. Our method successfully reconstructed all cells and vasculature within a large OMLIT dataset, enabling basic morphological classification and analysis. We also show that this approach can perform effectively on thicker sections, extending its applicability to sub-micron scale slices, saving sample preparation and imaging time, and increasing imaging throughput. Consequently, this method emerges as a promising candidate for high-speed, high-throughput brain tissue reconstruction and analysis. Our findings open new avenues for exploring the structure and function of the brain using OMLIT images.
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spelling pubmed-102165902023-05-27 A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping Wang, Tianyi Shi, Peiyao Luo, Dingsan Guo, Jun Liu, Hui Yuan, Jinyun Jin, Haiqun Wu, Xiaolong Zhang, Yueyi Xiong, Zhiwei Zhu, Jinlong Zhou, Renjie Zhang, Ruobing Brain Sci Article The mammalian brain, with its complexity and intricacy, poses significant challenges for researchers aiming to understand its inner workings. Optical multilayer interference tomography (OMLIT) is a novel, promising imaging technique that enables the mapping and reconstruction of mesoscale all-cell brain atlases and is seamlessly compatible with tape-based serial scanning electron microscopy (SEM) for microscale mapping in the same tissue. However, currently, OMLIT suffers from imperfect coatings, leading to background noise and image contamination. In this study, we introduced a new imaging configuration using carbon spraying to eliminate the tape-coating step, resulting in reduced noise and enhanced imaging quality. We demonstrated the improved imaging quality and validated its applicability through a correlative light–electron imaging workflow. Our method successfully reconstructed all cells and vasculature within a large OMLIT dataset, enabling basic morphological classification and analysis. We also show that this approach can perform effectively on thicker sections, extending its applicability to sub-micron scale slices, saving sample preparation and imaging time, and increasing imaging throughput. Consequently, this method emerges as a promising candidate for high-speed, high-throughput brain tissue reconstruction and analysis. Our findings open new avenues for exploring the structure and function of the brain using OMLIT images. MDPI 2023-04-24 /pmc/articles/PMC10216590/ /pubmed/37239183 http://dx.doi.org/10.3390/brainsci13050711 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wang, Tianyi
Shi, Peiyao
Luo, Dingsan
Guo, Jun
Liu, Hui
Yuan, Jinyun
Jin, Haiqun
Wu, Xiaolong
Zhang, Yueyi
Xiong, Zhiwei
Zhu, Jinlong
Zhou, Renjie
Zhang, Ruobing
A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title_full A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title_fullStr A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title_full_unstemmed A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title_short A Convenient All-Cell Optical Imaging Method Compatible with Serial SEM for Brain Mapping
title_sort convenient all-cell optical imaging method compatible with serial sem for brain mapping
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10216590/
https://www.ncbi.nlm.nih.gov/pubmed/37239183
http://dx.doi.org/10.3390/brainsci13050711
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