Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy

Min Kyo Jung; Ji Young Mun

doi:10.3791/56482

JoVE Journal
Biology

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JoVE Journal Biology

Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy

샘플 준비 및 전송 전자 현미경 검사 법에 의해 Exosomes의 영상

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11:15 min

January 4, 2018

DOI: 10.3791/56482-v

Min Kyo Jung¹, Ji Young Mun²

¹Department of Convergence Medicine, University of Ulsan College of Medicine and Asan Institute for Life Sciences,Asan Medical Center, ²Synaptic Circuit Plasticity Laboratory, Department of Structure and Function of Neural Network,Korea Brain Research Institute

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Overview

This protocol outlines techniques for transmission electron microscopy, focusing on the morphology of purified extracellular vesicles and protein localization. It aims to enhance understanding of exosome categorization and the specific proteins they contain.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Extracellular Vesicle Research

Background

Extracellular vesicles play a crucial role in cell communication.
Understanding their structure and protein content is vital for disease diagnosis.
Electron microscopy provides detailed insights into vesicle morphology.
Specific protein localization can inform on their functional roles.

Purpose of Study

To observe the morphology of extracellular vesicles.
To localize specific proteins within these vesicles.
To contribute to the understanding of exosome biology in health and disease.

Methods Used

Transmission electron microscopy techniques.
Negative staining for visualization.
Ultrathin sectioning for detailed structural analysis.
Immuno-gold labeling for specific protein localization.

Main Results

Detailed morphology of purified extracellular vesicles observed.
Localization of specific proteins within and outside exosomes achieved.
Insights gained into the role of exosomes in disease contexts.
Methodology applicable to various biological samples.

Conclusions

Transmission electron microscopy is effective for studying extracellular vesicles.
Specific protein localization enhances understanding of exosome functions.
This technique has implications for diagnosing diseases like cancer.

Frequently Asked Questions

What is the main goal of this protocol?

The main goal is to observe the morphology of extracellular vesicles and localize specific proteins within them.

What techniques are used in this study?

Techniques include transmission electron microscopy, negative staining, ultrathin sectioning, and immuno-gold labeling.

Why is understanding exosomes important?

Understanding exosomes is crucial for insights into cell communication and disease diagnosis.

What challenges might new users face?

New users may struggle with contamination and non-specific binding of immuno-gold particles.

Can this method be applied to other samples?

Yes, the methodology can be applied to various biological samples beyond exosomes.

이 프로토콜 전송 전자 현미경 검사 법이 부정적인 얼룩이 지기, 상세한 구조 및 면역-골드 라벨 exosomes에 특정 단백질의 위치를 확인 하려면 ultrathin 단면에 필요한 다양 한 기법을 설명 합니다.

이 절차의 전반적인 목표는 정제된 세포외 소포체의 형태를 관찰하고 전자 현미경을 사용하여 세포외 소포체 내에서 특정 단백질 국소화를 수행하는 것입니다. 이 방법은 세포외 소포체 연구 기간의 주요 질문, 예를 들어 엑소좀 및 그 내부에 위치한 특정 단백질의 분류와 같은 질문에 답하는 데 도움이 될 수 있습니다. 이 기술의 가장 큰 장점은 엑소좀 내부와 외부에 위치한 특정 단백질의 부분을 관찰하는 데 사용할 수 있다는 것입니다.

이 기술의 의미는 암 및 박테리아 감염을 포함한 여러 질병의 진단을 보장하는 것입니다. 이 방법은 엑소좀에 대한 통찰력을 제공할 수 있지만 다른 생물학적 샘플, 미세세포 배양 및 특정 단백질 국소화에도 적용할 수 있습니다. 일반적으로 이 방법을 처음 사용하는 개인은 면역금 입자의 비특이적 결합과 같은 오염을 일으키기 때문에 어려움을 겪을 것입니다.

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