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Super-resolution Fluorescence Microscopy01:37

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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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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
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高通量单分子显微镜具有可适应的空间分辨率,使用可交换的寡核酸标签.

Klarinda de Zwaan1, Ran Huo1, Myron N F Hensgens2

  • 1Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 2629 HZ Delft, The Netherlands.

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概括
此摘要是机器生成的。

这项研究引入了一个更快的生物成像工作流程,结合了超分辨率光学波动成像 (SOFI) 和DNA-PAINT显微镜. 这种方法使细胞结构成像加速30-300倍,提高了生物分子研究的吞吐量.

关键词:
这就是DNA-PAINT.闪的动力学 闪的动力学光波动成像技术 光波动成像高通量显微镜的显微镜.单分子局部化显微镜.超高分辨率成像成像技术

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科学领域:

  • 生物物理学的生物物理.
  • 细胞生物学 细胞生物学
  • 显微镜的使用方法

背景情况:

  • 超分辨率显微镜可以提供分子级别的可视化,但经常受到低吞吐量的影响.
  • 单分子定位显微镜 (SMLM) 技术是可访问的,但对于大规模查的速度有限.
  • 目前的局限性阻碍了先进显微镜在高通量生物分子研究中的应用.

研究的目的:

  • 开发一个工作流程,以有效地收集超分辨率显微镜中的数据.
  • 为了提高DNA-PAINT显微镜的吞吐量,用于细胞成像应用.
  • 根据实验需求,可灵活调整成像分辨率和采集时间.

主要方法:

  • 使用DNA-PAINT探针利用DNA的Oligo杂交动力学,进行量身定制的光闪.
  • 集成快速波动成像 (SOFI) 使用单分子定位显微镜 (DNA-PAINT).
  • 开发了一个混合工作流,从大面积扫描开始,然后是有针对性的SMLM.

主要成果:

  • 与传统的DNA-PAINT相比,实现了细胞结构的30-300倍更快的成像.
  • 在4分钟内证明了大面积 (0.65mm × 0.52mm) 的高通量SOFI成像.
  • 展示了高通量SOFI和高分辨率DNA-PAINT之间的灵活切换.

结论:

  • 结合DNA-PAINT和SOFI,提供了一种可适应生物成像分辨率和速度的多功能方法.
  • 拟议的工作流显著提高了SMLM技术的吞吐量.
  • 这种方法对复合和3D超分辨率成像的应用具有前景.