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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

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: Sep 13, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

单分子定位显微镜的误差取决于传感器,并且比理论预测的要大.

Alfonso Brenlla¹, Laila Deen¹, Paolo Annibale¹

¹School of Physics and Astronomy, University of St Andrews, St Andrews, United Kingdom.

Biophysical reports

|July 27, 2025

概括

随机局部化显微镜精确地确定单个分子的位置. 新方法揭示了理论精度估计往往低估了实验结果的一半,提供了一个软件独立的准确度指标.

科学领域:

显微镜的使用方法
生物物理学的生物物理.
纳米技术纳米技术

背景情况:

自2006年以来,随机定位显微镜 (SLM) 在亚衍射极限成像方面迅速发展.
SLM依赖于精确的单分子定位,这取决于光子收集和相机像素大小.
理论精度估计,如克莱默-拉奥下界,指导实验设计.

研究的目的:

在随机定位显微镜中评估理论精度估计的准确性.
将理论预测与实验定位精度进行比较.
提供一种可靠的方法来确定SLM中的本地化错误.

主要方法:

重温在随机定位显微镜中的关键研究.
分析理论和实验定位精度之间的差异.
开发一种软件独立的指标,用于实验性设置校准.

主要成果:

理论精度估计经常低估了实验本地化精度的约2倍.
现有的理论模型不能完全捕捉实验现实.
一个新的指标允许设置特定的定位定位错误.

结论:

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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

相关实验视频

Last Updated: Sep 13, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

对于SLM的理论精度计算需要改进以与实验结果相匹配.
准确评估局部化误差对于可靠的亚衍射成像至关重要.
提供的指标为校准SLM设置提供了一个实用的解决方案.