Epon Post Embedding Correlative Light and Electron Microscopy

Shuyuan Wang; Haiyan Xiong; Qiyuan Chang; Xudong Zhuang; Yaochen Wu; Xinrui Wang; Congxian Wu; Zhifei Fu

doi:10.3791/66141

Novel Protocol for Simultaneous Fluorescence and Ultrastructure Preservation

January 12, 2024

Epon Post indlejring korrelativt lys og elektronmikroskopi

00:46 min

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Ultratynd sektionering til epon post-embedding korrelativt lys og elektronmikroskopi (CLEM)

00:53 min

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Oprettelse af navigationskort til CLEM og forberedelse af de ultratynde sektioner til elektronmikroskopi

02:38 min

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Billedanalyse af Brightfield og fluorescens Billeder af musehjernesektionerne til epon post-indlejring korrelativt lys og elektronmikroskopi

02:13 min

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Registrering af fluorescensmikroskopibilledet med elektronmikroskopibilledet for korrelativt lys og elektronmikroskopi

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

Vi præsenterer en detaljeret protokol for Epon post-embedding korrelativt lys og elektronmikroskopi ved hjælp af et fluorescerende protein kaldet mScarlet. Denne metode kan opretholde fluorescensen og ultrastrukturen samtidigt. Denne teknik er egnet til en lang række biologiske anvendelser.