Novel Protocol for Simultaneous Fluorescence and Ultrastructure Preservation

January 12, 2024

Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation

00:46 min

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Ultrathin Sectioning for Epon Post-Embedding Correlative Light and Electron Microscopy (CLEM)

00:53 min

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Creating the Navigation Map for CLEM and Preparation of the Ultrathin Sections for Electron Microscopy

02:38 min

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Image Analysis of Brightfield and Fluorescence Images of the Mouse Brain Sections for Epon Post-Embedding Correlative Light and Electron Microscopy

02:13 min

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Registration of the Fluorescence Microscopy Image with the Electron Microscopy Image for Correlative Light and Electron Microscopy

02:30 min

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Shuyuan Wang*¹, Haiyan Xiong*^1,2, Qiyuan Chang*^1,3, Xudong Zhuang*^4,5,6, Yaochen Wu³, Xinrui Wang^5,6, Congxian Wu, Zhifei Fu^7,8

¹Public Technology Service Center,Fujian Medical University, ²The School of Pharmacy,Fujian Medical University, ³The School of Basic Medical Sciences,Fujian Medical University, ⁴College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics,Fujian Medical University, ⁵Medical Research Center,Fujian Maternity and Child Health Hospital, ⁶NHC Key Laboratory of Technical Evaluation of Fertility Regulation for Non-human Primate,Fujian Maternity and Child Health Hospital, ⁷Institute of Neuroscience,Fujian Medical University, ⁸Fujian Key Laboratory of Molecular Neurology,Fujian Medical University

Summary

We present a detailed protocol for Epon post-embedding correlative light and electron microscopy using a fluorescent protein called mScarlet. This method can maintain the fluorescence and the ultrastructure simultaneously. This technique is amenable to a wide variety of biological applications.