Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands

Isabella Finco; Gary D. Hammer

doi:10.3791/58530

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands

小鼠肾上腺的分离、固定和免疫荧光成像

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08:37 min

October 2, 2018

DOI: 10.3791/58530-v

Isabella Finco¹, Gary D. Hammer^1,2,3,4,5

¹Department of Internal Medicine (Division of Metabolism, Endocrinology, and Diabetes),University of Michigan Health System, ²Department of Cell and Developmental Biology,University of Michigan Health System, ³Department of Molecular and Integrative Physiology,University of Michigan Health System, ⁴Endocrine Oncology Program,University of Michigan Health System, ⁵Comprehensive Cancer Center,University of Michigan Health System

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本文提出了一种从小鼠中分离肾上腺的方法, 修复组织, 切片, 并进行免疫荧光染色。

免疫荧光是一种基本方法，可以帮助回答肾上腺领域有关肾上腺区、细胞特性和功能的关键问题。这项技术的主要优点是，它允许研究多种蛋白质，同时保持肾上腺组织的形态。要去除粘液肾上腺，切开周围的脂肪组织，将腺体放在含有新鲜 PBS 的 24 多井板的一个井中。

在解剖显微镜下将腺体转移到培养皿或玻璃支撑上，并使用两根 25 量针去除相邻的脂肪组织。当所有的脂肪都去除后，在4摄氏度下将组织用4%的半甲醛固定，摇动两个小时。在孵育结束时，在4摄氏度下用3个15分钟洗涤腺体，每次洗涤一至两毫升新鲜PBS。

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免疫学和感染问题 140 肾上腺免疫荧光小鼠组织固定石蜡切片