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成年斑马鱼脊髓结构的整装标本制备、透明处理及可视化方案。

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

作者信息

Rebocho da Costa Mariana, Ribeiro Ana, Quitéria Raquel, Neves-Silva Dalila, Saúde Leonor

机构信息

Instituto 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.

Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, 1649-028 Lisboa, Portugal.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103491. doi: 10.1016/j.xpro.2024.103491. Epub 2024 Dec 9.

Abstract

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..

摘要

血管修复是脊髓损伤反应的关键要素,但在斑马鱼研究中一直被显著忽视。在这里,我们详细介绍了斑马鱼全脊髓的提取和解剖,以及用于可视化脊髓结构(如血管网络)的优化全组织免疫荧光染色和透明化方案。这些样本的其他下游应用,从明胶包埋到冷冻切片和组织染色,可在透明化逆转后进行,本研究也对此进行了描述。有关本方案使用和执行的完整详细信息,请参考里贝罗等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05ca/11683226/8172fbad9cda/fx1.jpg

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