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

Epon Post Встраивание коррелятивной световой и электронной микроскопии

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Biology

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Epon Post Embedding Correlative Light and Electron Microscopy

Epon Post Встраивание коррелятивной световой и электронной микроскопии

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08:47 min

January 12, 2024

DOI: 10.3791/66141-v

Shuyuan Wang*¹, Haiyan Xiong*^1,2, Qiyuan Chang*^1,3, Xudong Zhuang*^4,5,6, Yaochen Wu^1,3, Xinrui Wang^4,5,6, Congxian Wu¹, Zhifei Fu^1,7,8

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

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Overview

This study presents a detailed protocol for Epon post-embedding correlative light and electron microscopy using the fluorescent protein mScarlet. The method successfully preserves both fluorescence and ultrastructure, making it applicable to various biological applications.

Key Study Components

Research Area

Correlative light and electron microscopy
Fluorescent protein applications
Microscopy techniques

Background

Combining fluorescent microscopy with electron microscopy
Challenges in preserving fluorescence during preparation
Use of mScarlet for enhanced imaging

Methods Used

Correlative light and electron microscopy techniques
Mouse brain tissue as the biological model
Post-embedding using Epon and mScarlet

Main Results

Successful preservation of fluorescence and ultrastructure
Detailed imaging of brain tissue
Effective alignment of fluorescence and electron microscopy signals

Conclusions

The study demonstrates a reliable method for correlative light and electron microscopy
Highlights the potential for broader applications in biology research

Frequently Asked Questions

What is the main advantage of using mScarlet?

mScarlet allows for the simultaneous preservation of fluorescence and ultrastructure in microscopy.

What biological materials were used in this protocol?

Mouse brain tissue expressing the fluorescent protein mScarlet was used.

Can this method be applied to other proteins?

While the protocol focuses on mScarlet, it may be adaptable to other fluorescent proteins.

How do you ensure proper alignment of images in this method?

Images are aligned using specific software during the registration process to match fluorescence and electron microscopy signals.

What is Epon and why is it used?

Epon is a resin used in embedding samples for electron microscopy, helping to preserve fine cellular structures.

What are the main applications of this microscopy technique?

It can be used in various biological research applications, including cell biology and developmental studies.

Is this method suitable for all types of tissues?

The protocol is optimized for mouse brain tissue but may be adaptable to other tissues with proper modifications.

Мы представляем подробный протокол послевстраивающей коррелятивной световой и электронной микроскопии Epon с использованием флуоресцентного белка mScarlet. Этот метод позволяет поддерживать флуоресценцию и ультраструктуру одновременно. Эта методика поддается широкому спектру биологических применений.

Коррелятивная световая и электронная микроскопия - это полностью стабильный метод микроскопии, который может сочетать информацию о локализации, предоставляемую флуоресцентной микроскопией, и является ультраструктурой электронной микроскопии. Тем не менее, менталитет этой техники заключается в том, чтобы сохранить флуоресценцию после того, как у вас есть простая подготовка. Используя этот флуоресцентный белок, мы можем получить коррелятивный свет и электронный микроскоп, который может одновременно поддерживать флуоресценцию и ультраструктуру.

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