相关实验视频

Updated: Jun 8, 2025

Direct Stochastic Optical Reconstruction Microscopy of Extracellular Vesicles in Three Dimensions

Direct Stochastic Optical Reconstruction Microscopy of Extracellular Vesicles in Three Dimensions

Published on: August 26, 2021

在单个EV级别使用传输电子显微镜分析细胞外囊相关DNA

Thupten Tsering^1,2, Amélie Nadeau^1,2, Janusz Rak^1,3

¹Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada.

Current protocols

|November 8, 2024

概括

新的协议可以准确检测细胞外囊DNA (EV-DNA),这是液体活检的一个有希望的生物标志物. 这些方法利用传输电子显微镜在各种生物样本中进行详细的EV-DNA分析.

科学领域:

细胞生物学细胞生物学
生物化学生物化学
生物标志物发现发现

背景情况:

细胞外囊泡 (EVs) 通过载荷转移 (包括DNA) 调解细胞与细胞的通信.
EV-关联DNA (EV-DNA) 是一种潜在的液体活检生物标志物,但由于EV的纳米结构和低丰度,分析其具有挑战性.
准确的EV-DNA检测协议对于理解其生理和病理作用至关重要.

研究的目的:

开发和验证用于精确检测和表征EV-DNA的新型协议.
确认细胞外囊泡内双链DNA的存在和定位.
建立使用先进显微镜识别与EV相关的DNA生物标记物的方法.

主要方法:

协议1:将EV的超薄切割与免疫黄金标记和传输电子显微镜 (TEM) 结合起来,用于检测光线dDNA.
协议2:采用全套EV免疫黄金标签,用于形态分析和表面相关DNA检测.
使用TEM成像来高分辨率可视化EV结构和DNA定位.

主要成果:

通过使用TEM在癌细胞衍生的EV上同时检测dSDNA和EV表面蛋白tetraspanin 9.
验证了EV-DNA在EV光层和EV表面的存在.
建立了一种用于标记内部和表面EV蛋白的多功能方法.

结论:

开发的协议提高了EV-DNA的准确检测和表征.
这些方法有助于单个EV分析和确定与EV相关的生物标志物.
这些发现支持了EV-DNA作为液体活检和疾病诊断中的生物标记物的潜力.

关键词:

双链DNA是双链DNA的组成部分.细胞外囊泡中的细胞外囊泡.免疫黄金标签的标签泰特拉斯帕宁 9 9 的使用.传输电子显微镜的使用

更多相关视频

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Published on: January 7, 2019

Freeze-Fracture Electron Microscopy for Extracellular Vesicle Analysis

Freeze-Fracture Electron Microscopy for Extracellular Vesicle Analysis

Published on: September 16, 2022

相关概念视频

Electron Microscope Tomography and Single-particle Reconstruction

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

Transmission Electron Microscopy

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...