象限暗场用于细胞内点点的无标签成像
Tarek E Moustafa1, Rachel L Belote2,3, Edward R Polanco1
1University of Utah, Department of Chemical Engineering, Salt Lake City, Utah, United States.
Journal of biomedical optics
|December 2, 2024
概括
我们开发了一种新的无标签成像技术,称为象限暗场 (QDF) 显微镜. QDF显微镜通过消除边缘信号,有效地可视化活细胞中的亚细胞结构,改善有机体量化.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜的使用方法
- 生物光子学 生物光子学
背景情况:
- 无标签成像对于研究细胞行为而没有人工制造物至关重要.
- 传统的暗场显微镜与隐藏细胞内细节的边缘信号作斗争.
研究的目的:
- 引入象限暗场 (QDF) 成像,以免标签对亚细胞结构进行可视化.
- 能够使小细胞特征从活细胞中较大的结构中分离出来.
主要方法:
- 使用可编程LED阵列进行定向照明.
- 改变照明方向以编码基于光散射的特征尺寸信息.
- 开发了一种计算方法,以光学消除暗场图像中的边缘信号.
主要成果:
- QDF成功地解决了小细胞特征,而不会受到较大的结构的干扰.
- 与传统的暗场相比,QDF成像在细胞形状变化期间提供了更一致的信号.
- QDF信号与流动细胞计侧分散相关,通过有机细胞含量区分细胞.
结论:
- QDF成像增强了活细胞中亚细胞结构的研究.
- 与无标签暗场显微镜相比,它可以更好地量化有机细胞含量.
- 可以与其他技术相结合,如用于多维细胞分析的定量相位成像.
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...
01:20Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.