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相关概念视频

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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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,...
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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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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...
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Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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...
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相关实验视频

Updated: May 26, 2025

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
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快速重建和光学分割的三维结构化照明显微镜.

Ruijie Cao1,2, Yaning Li1,2,3, Wenyi Wang1,2,4

  • 1Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China.

Innovation (Cambridge (Mass.))
|February 24, 2025
PubMed
概括

我们开发了快速的3D结构化照明显微镜 (3DSIM),称为FO-3DSIM. 这种方法显著加快了重建的速度,使近实时,大视野超高分辨率成像与减少光损伤.

关键词:
大视野成像成像大视野成像光学切割是指光学切割的方法.实时观察实时观察实时观察重建速度的重建速度.三维结构照明照明三维结构照明

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Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy f3D-SIM
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Three and Four-Dimensional Visualization and Analysis Approaches to Study Vertebrate Axial Elongation and Segmentation
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相关实验视频

Last Updated: May 26, 2025

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Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy f3D-SIM
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Three and Four-Dimensional Visualization and Analysis Approaches to Study Vertebrate Axial Elongation and Segmentation
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科学领域:

  • 显微镜和成像技术的发展.
  • 生物物理学和细胞成像技术
  • 超高分辨率显微镜的使用方法

背景情况:

  • 传统的三维结构化照明显微镜 (3DSIM) 提供3D超分辨率,但受重建时间缓慢的影响,限制了高通量应用.
  • 传统的3DSIM需要多个z层,对焦外光敏感,阻碍更厚样本的成像,并引入文物.
  • 在2D SIM和6层3DSIM之间存在很大的差距,限制了对生物结构的观察.

研究的目的:

  • 开发一种新的3DSIM方法,克服缓慢重建和对失焦背景敏感性的局限性.
  • 为了提高生物样本3D超分辨率成像的速度和效率.
  • 为了实现近实时,大视野的3D超分辨率成像,减少光损伤.

主要方法:

  • 将空间域重建与光学分割SIM集成在一起,创建FO-3DSIM.
  • 实施有限的z层采集策略,以实现更快的3D重建.
  • 使用高保真,低光子重建算法,建立在之前的Open-3DSIM进步之上.

主要成果:

  • 与传统方法相比,FO-3DSIM实现了高达855.7倍的重建速度.
  • 在有限的z层和高度失焦的背景条件下表现出卓越的性能.
  • 成功成像大视野 (0.453 x 0.453毫米) 的小鼠动素与3D超分辨率在40分钟以下 (获取+重建).
  • 通过仅使用三个z层,实现了实时直播actin动态的近实时成像,并观察了ER管.

结论:

  • FO-3DSIM显著加速3D超分辨率显微镜,使其适合高通量和近实时应用.
  • 该方法有效地减少了重建时间和光损伤,扩大了成像更厚样本和动态过程的范围.
  • FO-3DSIM为先进的6D成像 (xyz,多色,时间,极化) 铺平了道路,提高了速度和减少了文物.