Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures

Summary: The vascular repair, a pivotal element of the spinal cord (SC) injury response, has been notably neglected in zebrafish research. Here, we detail the extraction and dissection of whole-mount zebrafish SCs and the optimized whole-mount immunofluorescence staining and clearing protocols for t...

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Main Authors: Mariana Rebocho da Costa, Ana Ribeiro, Raquel Quitéria, Dalila Neves-Silva, Leonor Saúde
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006567
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author Mariana Rebocho da Costa
Ana Ribeiro
Raquel Quitéria
Dalila Neves-Silva
Leonor Saúde
author_facet Mariana Rebocho da Costa
Ana Ribeiro
Raquel Quitéria
Dalila Neves-Silva
Leonor Saúde
author_sort Mariana Rebocho da Costa
collection DOAJ
description Summary: The vascular repair, a pivotal element of the spinal cord (SC) injury response, has been notably neglected in zebrafish research. Here, we detail the extraction and dissection of whole-mount zebrafish SCs and the optimized whole-mount immunofluorescence staining and clearing protocols for the visualization of SC structures, such as the vascular network. Additional downstream applications of these samples, from gelatin embedding to cryosectioning and tissue staining, can be done after clearing reversion and are also described in this work.For complete details on the use and execution of this protocol, please refer to Ribeiro et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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spelling doaj-art-a3e75493502d410d887880713bd2ce8a2025-08-20T02:39:03ZengElsevierSTAR Protocols2666-16672024-12-015410349110.1016/j.xpro.2024.103491Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structuresMariana Rebocho da Costa0Ana Ribeiro1Raquel Quitéria2Dalila Neves-Silva3Leonor Saúde4Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal; GIMM - Gulbenkian Institute for Molecular Medicina, Avenida Prof. Egas Moniz, 1649-035 Lisboa, Portugal; Corresponding authorInstituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, PortugalInstituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, PortugalInstituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal; GIMM - Gulbenkian Institute for Molecular Medicina, Avenida Prof. Egas Moniz, 1649-035 Lisboa, PortugalInstituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal; Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal; GIMM - Gulbenkian Institute for Molecular Medicina, Avenida Prof. Egas Moniz, 1649-035 Lisboa, Portugal; Corresponding authorSummary: The vascular repair, a pivotal element of the spinal cord (SC) injury response, has been notably neglected in zebrafish research. Here, we detail the extraction and dissection of whole-mount zebrafish SCs and the optimized whole-mount immunofluorescence staining and clearing protocols for the visualization of SC structures, such as the vascular network. Additional downstream applications of these samples, from gelatin embedding to cryosectioning and tissue staining, can be done after clearing reversion and are also described in this work.For complete details on the use and execution of this protocol, please refer to Ribeiro et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724006567Cell BiologyMicroscopyModel OrganismsNeuroscience
spellingShingle Mariana Rebocho da Costa
Ana Ribeiro
Raquel Quitéria
Dalila Neves-Silva
Leonor Saúde
Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
STAR Protocols
Cell Biology
Microscopy
Model Organisms
Neuroscience
title Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
title_full Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
title_fullStr Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
title_full_unstemmed Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
title_short Protocol for whole-mount preparation, clearing, and visualization of the adult zebrafish spinal cord structures
title_sort protocol for whole mount preparation clearing and visualization of the adult zebrafish spinal cord structures
topic Cell Biology
Microscopy
Model Organisms
Neuroscience
url http://www.sciencedirect.com/science/article/pii/S2666166724006567
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