相对光和电子显微镜在受控环境中的定义细胞周期阶段
Helena Bragulat-Teixidor1, Shotaro Otsuka2
1Max Perutz Labs, Vienna Biocenter Campus (VBC), Vienna, Austria; Vienna BioCenter PhD Program, Doctoral School of the University of Vienna and Medical University of Vienna, Vienna, Austria; Medical University of Vienna, Center for Medical Biochemistry, Vienna, Austria.
Methods in cell biology
|May 5, 2024
概括
这项研究引入了一种新的协议,用于在近原生条件下观察细胞中的动态细胞内变化. 这种方法可以对细胞结构进行高分辨率成像,有助于了解细胞生物学和发育.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜的使用方法
- 生物物理学的生物物理.
背景情况:
- 细胞在对刺激的反应中表现出动态的结构变化.
- 在高分辨率下了解这些动态对于机械洞察至关重要.
- 目前的方法可能无法在观察期间完全保留本地细胞条件.
研究的目的:
- 提出一种用于现场研究动态细胞内结构的新方案.
- 在接近本土条件下实现高时空分辨率.
- 为了使细胞动态的机械学理解.
主要方法:
- 开发一个专门的细胞培养,运输和成像室.
- 室内的环境控制 (温度,气体度).
- 该协议的应用用于研究哺乳动物细胞周期期间的超结构变化.
主要成果:
- 对动态细胞过程的现场成像协议的演示.
- 用高分辨率量化细胞周期期间的超结构变化.
- 环境控制系统的验证,以保持接近本土的条件.
结论:
- 开发的协议使得高分辨率,细胞动态的实地研究.
- 环境控制是保持原生条件以准确观察的关键.
- 该方法适用于原始细胞,组织和有机体.
相关概念视频
01:26Phase Contrast and Differential Interference Contrast Microscopy
Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
01:25Overview of Electron Microscopy
The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
01:22Overview of Microscopy Techniques
The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
01:29Two-Dimensional Microscopy in Microbiology
Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...