Abstract：
Immunofluorescence, a transformative tool in cellular biology, is employed to dissect the intricate mechanisms of cholesterol trafficking in human reproductive tissues. Autophagy, a key player in cellular homeostasis, particularly lipophagy, emerges as a free cholesterol source for steroidogenesis. In this chapter, we describe a comprehensive immunofluorescence staining protocol, with details provided for the precise visualization of subcellular dynamics of mitochondria, lysosomes, and lipid droplets in ex vivo testicular tissue and primary luteal granulosa cell culture models, pivotal components in sex steroid biosynthesis. Here, we detail the culture, treatment, and immunofluorescence protocols, providing a comprehensive guide for researchers. The provided immunofluorescence toolkit serves as a valuable resource for researchers, paving way for advancements in human reproductive health to investigate the intricate interplay between autophagy, lipophagy, and cholesterol trafficking.
摘要：
免疫荧光,细胞生物学中的转化工具，用于剖析人类生殖组织中胆固醇贩运的复杂机制。自噬,细胞稳态的关键角色，特别是吸脂症，作为类固醇生成的游离胆固醇来源出现。在这一章中,我们描述了一个全面的免疫荧光染色方案，为线粒体亚细胞动力学的精确可视化提供了细节，溶酶体,和离体睾丸组织和原代黄体颗粒细胞培养模型中的脂滴，性类固醇生物合成的关键成分。这里,我们详细介绍了文化，治疗,和免疫荧光协议，为研究人员提供全面的指导。提供的免疫荧光工具包是研究人员的宝贵资源，为人类生殖健康的进步铺平了道路，以研究自噬之间的复杂相互作用，吸脂症,胆固醇贩运。
