Department of Neurological Surgery, The Ohio State University, Columbus, OH 43210, USA.
STAR Protoc. 2022 Jan 19;3(1):101109. doi: 10.1016/j.xpro.2021.101109. eCollection 2022 Mar 18.
Here we provide a clearing-free protocol for processing intact, whole mount subcutaneous white adipose tissue (scWAT) for immunofluorescence as an alternative to current clearing-based approaches. We use a combination of Z-depth reduction and autofluorescence quenching techniques to fluorescently label, image, and quantify adipose tissue innervation effectively throughout intact mouse tissues without the need for optical clearing or light sheet microscopy. This protocol has been optimized and validated for adipose neurovascular labeling. For complete details on the use and execution of this protocol, please refer to Willows et al. (2021).
在这里,我们提供了一种无需透明化处理的方案,用于处理完整的、未经处理的皮下白色脂肪组织(scWAT)的免疫荧光,以替代当前基于透明化的方法。我们使用 Z 深度减少和自发荧光猝灭技术的组合,有效地对完整的小鼠组织中的脂肪组织神经支配进行荧光标记、成像和定量,而无需光学透明化或光片显微镜。该方案已针对脂肪神经血管标记进行了优化和验证。有关该方案使用和执行的完整详细信息,请参阅 Willows 等人(2021 年)。