Free-floating Immunostaining of Mouse Brains.
Free-floating Immunostaining of Mouse Brains.
复制标题
小鼠脑的自由浮动免疫染色。
DOI：
10.3791/62876
复制
发表时间：
2021-10-07
期刊：
影响因子：
--
通讯作者：
Xu Y
中科院分区：
文献类型：
--
作者：
Tu L;Zhang N;Conde KM;Bean JC;Wang C;Xu Y
小鼠脑的免疫组织化学染色是神经科学中常用的常规技术，用于研究能量代谢和其他神经生物学过程调节的中枢机制。然而，脑组织学结果的质量、可靠性和可重复性可能因实验室而异。对于每个染色实验，需要根据物种、组织、靶蛋白和试剂工作条件的差异优化关键步骤。本文详细介绍了一个可靠的工作流程，包括主动脉内灌注，脑切片，自由浮动免疫染色，组织安装和成像，这可以很容易地遵循该领域的研究人员。还讨论了如何修改这些程序，以满足研究人员的个性化需求。为了说明该方案的可靠性和有效性，在小鼠脑中用生物素标记的紫藤凝集素（WFA）和精氨酸加压素（AVP）与抗AVP抗体对神经元周围网进行染色。最后，整个过程的关键细节已经得到解决，该协议的优点相比，其他协议。综上所述，本文提出了一种优化的方案，小鼠脑组织的自由浮动免疫染色。遵循此方案使初级和高级科学家更容易进行此过程，以提高免疫染色研究的质量，可靠性和重现性。
Immunohistochemical staining of mouse brains is a routine technique commonly used in neuroscience to investigate central mechanisms underlying the regulation of energy metabolism and other neurobiological processes. However, the quality, reliability, and reproducibility of brain histology results may vary among laboratories. For each staining experiment, it is necessary to optimize the key procedures based on differences in species, tissues, targeted proteins, and the working conditions of the reagents. This paper demonstrates a reliable workflow in detail, including intra-aortic perfusion, brain sectioning, free-floating immunostaining, tissue mounting, and imaging, which can be followed easily by researchers in this field. Also discussed are how to modify these procedures to satisfy the individual needs of researchers. To illustrate the reliability and efficiency of this protocol, perineuronal nets were stained with biotin-labeled Wisteria florbunda agglutinin (WFA) and arginine vasopressin (AVP) with an anti-AVP antibody in the mouse brain. Finally, the critical details for the entire procedure have been addressed, and the advantages of this protocol compared to those of other protocols. Taken together, this paper presents an optimized protocol for free-floating immunostaining of mouse brain tissue. Following this protocol makes this process easier for both junior and senior scientists to improve the quality, reliability, and reproducibility of immunostaining studies.
登录
查看更多内容
影响因子：
8.8
作者：
He Y;Shu G;Yang Y;Xu P;Xia Y;Wang C;Saito K;Hinton A Jr;Yan X;Liu C;Wu Q;Tong Q;Xu Y
通讯作者：
Xu Y
影响因子：
10.6
作者：
Xu, Pingwen;He, Yanlin;Xu, Yong
通讯作者：
Xu, Yong
DOI：
10.3791/3564
发表时间：
2012-07-30
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Gage, Gregory J;Kipke, Daryl R;Shain, William
通讯作者：
Shain, William
DOI：
10.1002/cne.25100
发表时间：
2021-06
期刊：
The Journal of comparative neurology
影响因子：
--
作者：
Liu H;He Y;Beck J;da Silva Teixeira S;Harrison K;Xu Y;Sisley S
通讯作者：
Sisley S
影响因子：
4.5
作者：
Wong SK;Chin KY;Suhaimi FH;Fairus A;Ima-Nirwana S
通讯作者：
Ima-Nirwana S