长期内生细胞亚细胞成像与共聚焦扫描光场显微镜
Zhi Lu1,2,3,4,5,6, Siqing Zuo1,2,3,4, Minghui Shi7
1Department of Automation, Tsinghua University, Beijing, China.
Nature biotechnology
|May 27, 2024
概括
同焦扫描光场显微镜 (csLFM) 克服了观察亚细胞动态的局限性. 这种新技术提供了高速,高分辨率的3D成像,减少了光毒性,并改善了信号与背景的比率.
科学领域:
- 生物物理学的生物物理.
- 显微镜的使用方法
- 细胞生物学 细胞生物学
背景情况:
- 长期的亚细胞动态观测受到背景光和组织散射的阻碍.
- 当前的共聚焦显微镜方法涉及平行化,分辨率和光毒性之间的权衡.
研究的目的:
- 引入共聚焦扫描光场显微镜 (csLFM) 以实现高保真,高速的3D成像.
- 克服现有的显微镜技术在观察亚细胞动态方面的局限性.
主要方法:
- 在扫描光场显微镜 (sLFM) 中,将轴延长的直线相聚焦照明与滚动快门集成.
- 同时进行3D激发和检测,使激发强度降低 (<1mW mm−2).
主要成果:
- 实现了接近衍射极限的分辨率,光学切割和低光毒性.
- 与sLFM相比,显示的信号与背景比率高15倍.
- 在各种物种和充满挑战的环境中,在超过25000个时间框架中,成功成像了细胞下动态.
结论:
- csLFM能够高准确性,高速3D成像亚细胞动态,减少光毒性.
- 该技术可提供高质量的大规模神经记录,减少交叉通话,有助于神经编码研究.
相关概念视频
Confocal Fluorescence Microscopy
13.2K
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,...
13.2K
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


