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相关概念视频

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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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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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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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实现实时重建大型视野单分子局部化显微镜,使用离散的场依赖点扩散函数.

Jun Lu1, Lei Xu1, Shuyao Liao1

  • 1Academy for Engineering and Technology, Yiwu Research Institute, Fudan University, Shanghai 200433, China.

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概括

光学偏差限制了3D单分子定位显微镜 (SMLM) 的视野. 本研究介绍了离散的取决于现场的点分布函数 (PSF),以实现在更大区域的精确定位,改进SMLM应用程序.

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科学领域:

  • 生物物理学的生物物理.
  • 光学显微镜的使用方法
  • 超高分辨率的成像技术

背景情况:

  • 单分子定位显微镜 (SMLM) 通过定位单个光分子来实现超分辨率.
  • 对于SMLM必不可少的高数值光圈目标显示出偏离轴的扭曲点传播函数 (PSF),限制视野 (FOV).
  • 这些异常引入了显著的3D定位错误,将SMLM限制在小FOV (例如50μm × 50μm) 上.

研究的目的:

  • 系统地研究光学偏差对大型FOV 3D SMLM的影响.
  • 提出和验证一种方法来扩大3D SMLM中的有效FOV.
  • 为了在大型成像区域中实现实时,高精度的分子定位.

主要方法:

  • 在不同的光学偏差下使用未经修改的,形的和双螺旋的PSF评估了定位准确性.
  • 开发并测试了离散的场依赖PSF用于偏差校正.
  • 采用GPU加速,用于模拟和生物样本的高速图像重建.

主要成果:

  • 光学偏差显著降低了FOV边缘的定位精度和准确性.
  • 离散的取决于场的PSF有效地纠正离轴偏差,使得在更大的区域中实现精确的3D定位.
  • 用GPU加速的离散PSF模型允许实时SMLM图像重建.

结论:

  • 光学偏差是大型FOV 3D SMLM的一个关键限制.
  • 独立的现场依赖PSF提供了一个可行的策略来克服这些局限性,增强FOV.
  • 使用GPU加速的实时重建使得大型FOV SMLM对生物研究更加实用.