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相关概念视频

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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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy iPALM
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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy iPALM

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量化图像扫描局部化显微镜的最小局部化不确定性.

Dylan Kalisvaart1, Shih-Te Hung1, Carlas S Smith1,2

  • 1Delft Center for Systems and Control, Delft University of Technology, Delft, the Netherlands.

Biophysical reports
|February 21, 2024
PubMed
概括

调制增强定位用于旋盘共聚焦显微镜 (SpinFlux) 提高了单分子定位精度. 这种技术为超分辨率显微镜提供了显著的进步,提高了生物成像中的定位精度.

科学领域:

  • 超高分辨率显微镜的使用方法
  • 光学成像技术的使用.
  • 生物物理学的生物物理.

背景情况:

  • 单分子定位显微镜 (SMLM) 实现了纳米级分辨率,但在精度上有局限性.
  • 调制增强的SMLM (meSMLM) 通过使用模式照明来连续激活发射器来提高精度.
  • 现有的meSMLM方法需要专门的设置,可能与所有显微镜平台不兼容.

研究的目的:

  • 介绍SpinFlux,一种用于旋盘共聚焦显微镜的新型调制增强定位方法.
  • 使用旋转磁盘对焦设置,提高SMLM的定位精度.
  • 将SpinFlux的性能与其他高级本地化技术进行比较.

主要方法:

  • SpinFlux利用一个旋转的磁盘,在照明和发射路径中有针孔,用于连续区域的照明.
  • 对每个模式进行强度调制的发射信号分析,以确定发射器位置.
  • 一个统计图像形成模型和克拉梅尔-拉奥下界被导出来量化定位不确定性.

主要成果:

  • 通过三种聚焦在发射器周围的照明模式,SpinFlux比SMLM实现了双倍的定位精度改进.
  • 使用四个甜甜圈形状的图案,SpinFlux显示了与SMLM相比的最大局部精度提升3.5.

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  • 在富里埃重量图像扫描显微镜重建上的定位提供了理想的全球精度改进 (比SMLM高2.1倍).

    • 结论:

    • 旋转Flux显著提高了旋转磁盘共聚焦显微镜中的定位精度.
    • 在超分辨率成像中,SpinFlux是用于局部精度改进的宝贵工具.
    • 图像扫描显微镜重建为全球精度改进提供了最大的潜力,而SpinFlux在本地改进方面表现出色.