相关实验视频
Updated: Jul 12, 2025

08:32
Conducting Multiple Imaging Modes with One Fluorescence Microscope
Published on: October 28, 2018
9.9K
光辐射波动使两光子超分辨率显微镜成为可能.
Motosuke Tsutsumi1,2, Taiga Takahashi1,2, Kentaro Kobayashi3
1Biophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, Japan.
Frontiers in cellular neuroscience
|October 26, 2023
概括
超分辨率辐射波动 (SRRF) 显微镜与双光子成像相结合,可实现深度大脑可视化. 这种新的2P-SRRF技术在体内实现了高空间分辨率和形态可重现性.
科学领域:
- 神经科学是一个神经科学.
- 显微镜的使用方法
- 生物物理学的生物物理.
背景情况:
- 深度超分辨率成像受限于厚型标本中的光散射.
- 超分辨率辐射波动 (SRRF) 通过依赖图像分析而不是光学操纵提供了一个潜在的解决方案.
- 现有的超分辨率技术在生物组织的透深度方面扎.
研究的目的:
- 将SRRF应用于双光子显微镜 (2P-SRRF) 进行深度超分辨率成像.
- 在空间分辨率,成像深度和形态可重现性方面描述2P-SRRF的性能.
- 为了证明2P-SRRF在脑部体内成像的可行性.
主要方法:
- 集成SRRF图像分析与两光子显微镜设置.
- 与结构化照明显微镜 (SIM) 相比,空间分辨率和形态复制性的表征.
- 在大脑模拟环境中测试2P-SRRF性能,深度超过几百微米.
- 在体内成像中优化SRRF处理参数.
主要成果:
- 2P-SRRF实现了与SIM相比的空间分辨率和形态重现性.
- 在几百微米以上的深度观察到空间分辨率的显著改善.
- 证明了脑皮质第五层体内高分辨率成像的成功.
- 该方法与现有的双光子显微镜兼容.
结论:
- 2P-SRRF克服了深度超分辨率成像的光学限制.
- 这种技术显著扩大了神经科学研究的可视化能力.
- 应用于双光子显微镜的SRRF代表了体内超分辨率研究的突破.
更多相关视频
相关概念视频
Super-resolution Fluorescence Microscopy
7.0K
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
7.0K
Protein Dynamics in Living Cells
2.1K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.1K
Total Internal Reflection Fluorescence Microscopy
5.8K
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
5.8K
Confocal Fluorescence Microscopy
13.3K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
13.3K

