Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing

小鼠眼部切片常规组织学、免疫荧光和RNA原位杂交多重检测的逐步制备方法

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作者:Ji Pang ,Nancy Thomas ,Dai Tsuchiya ,Tari Parmely ,Deyue Yan ,Ting Xie ,Yongfu Wang

Abstract

It can be challenging to maintain tissue integrity using established histology protocols. Here, we describe a protocol composed of Hartman's fixation, window technique, microwave-based tissue processing, optimized depigmentation, and antigen retrieval pretreatment. This is followed by the ViewRNA single-molecule fluorescence in situ hybridization and immunofluorescence techniques to optimize routine histological staining and molecular histology multiplexing assays. Our protocol is highly reproducible in any laboratory and may decrease animal usage and lab resource expenditure. For complete details on the use and execution of this protocol, please refer to Pang et al. (2021).

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