一种新的适用于活体斑马鱼成像观察的固定方法

陈二方 常会敏 安晓刚 邱建华 查定军

摘要 [目的] 建立一种更适用于活体斑马鱼成像的固定方法，为活体斑马鱼成像研究提供技术支持。[方法]通过比较4种不同的斑马鱼体位固定方法及成像观察效果，观察不同固定方法的可操作性和成像观察效果。[结果]使用6%甲基纤维素固定时，对斑马鱼进行拍照时容易产生摆动;使用0.05%MS-222固定时，斑马鱼很难保持合适的体位;使用1%的低熔点琼脂糖，相对费时; 使用6%甲基纤维素/0.05%MS-222固定时，既可以使斑马鱼保持湿润的环境，又可以保持理想的静止状态。[结论]通过比较4种固定方法发现，使用6%甲基纤维素/0.05%MS-222固定斑马鱼仔鱼可操作性最好，该方法可为斑马鱼活体研究提供良好的活体成像观察保障，有助于获取高质量的活体成像图片。

关键词 斑马鱼;活体成像;甲基纤维素;固定方法

中图分类号 S 917.4  文献标识码 A  文章编号 0517-6611（2021）23-0124-03

doi：10.3969/j.issn.0517-6611.2021.23.033

A New Fixation Method for in vivo Imaging and Observation of Zebrafish

CHEN Er-fang1， CHANG Hui-min2， AN Xiao-gang1 et al

（1.Otolaryngology，Head and Neck Surgery， Xijing Hospital， PLA Air Force Military Medical University，Xi’an， Shaanxi 710032; 2. Department of Otolaryngology，Head and Neck Surgery，The First Affiliated Hospital of Xi’an Medical University，Xi’an，Shaanxi 710077）

Abstract [Objective] To introduce a new fixation method for in vivo imaging of zebrafish， and provide technical supports for in vivo imaging research of zebrafish.[Method] Though comparing the position fixation methods and imaging observation effects of four different methods， the feasibility and imaging observation effects of different fixation methods were observed.[Result]6% methyl cellulose fixation method was used， it was easy to produce wobble in the process of taking pictures. When zebrafish was fixed with 0.05%MS-222， it was difficult to maintain the proper position.When 1% agarose with a low melting point was used， it was relatively time-consuming. When 6% methyl-cellulose /0.05%MS-222 were used for the fixation， it could not only keep zebrafish in a moist environment， but it could maintain an ideal static state. [Conclusion] By comparing four fixation methods， it was found that fixing zebrafish with 6% methylcellulose /0.05%MS-222 were the most maneuverable.This method could provide a good in vivo imaging observation guarantee for in vivo research of zebrafish， and it was helpful to obtain  in vivo imaging pictures with high-quality.

Key words Zebrafish;In vivo imaging;Methyl cellulose;Fixation method

基金项目 国家自然科学基金面上项目（81870732）;学科助推计划-先进学科建设项目（XJZT18X23）。

作者简介 陈二方（1984—），女，安徽宿州人，技师，硕士，从事耳聋的基础研究。通信作者，教授，博士，从事耳科基础与临床研究。

收稿日期 2021-07-28

斑马鱼作为一种热带鱼，胚胎发育速度快、养殖成本低、能大规模繁育[1-3]。研究发现，斑马鱼基因与人类基因的相似度在70%以上[4]，因此它受到生物学家的重视，已经成为最受重视的脊椎动物发育生物学研究的动物模型之一[5]。目前全球范围内有超过1 500个斑马鱼实验室，我国有300个以上的实验室利用斑马鱼开展相关试验[6]。……