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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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Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy
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使用商业可用的共聚焦显微镜进行传输强度相位成像.

Naru Yoneda1,2, Joe Sakamoto3,4, Takumi Tomoi5,6,7

  • 1Kobe University, Graduate School of System Informatics, Department of System Science, Kobe, Japan.

Journal of biomedical optics
|November 8, 2024
PubMed
概括

研究人员使用强度方程传输 (TIE) 将标准的共焦显微镜转换为相位测量显微镜. 这项创新使现有仪器能够进行定量相成像 (QPI),为细胞生物学提供新的见解.

关键词:
同焦点显微镜的共焦显微镜.光成像成像技术的使用.光显微镜的光显微镜.显微镜 显微镜是指使用显微镜.定量阶段成像成像技术运输强度方程的运输.

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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
  • 细胞生物学 细胞生物学

背景情况:

  • 对焦显微镜对于高分辨率的生物成像至关重要,特别是对于活细胞中的光.
  • 阶段信息提供了有价值的见解,包括物理参数测量和波面校正,但在商业对焦系统中缺少.
  • 现有的共聚焦显微镜缺乏集成相位成像能力.

研究的目的:

  • 适应商业上可用的共聚焦显微镜用于定量相位成像 (QPI).
  • 引入相位成像功能,而不需要对现有显微镜进行硬件修改.
  • 为了使生物学家能够使用他们目前的共聚焦显微镜设置来测量相位信息.

主要方法:

  • 使用强度方程运输 (TIE) 实现了相位成像.
  • 该方法利用标准共聚焦显微镜的明亮场成像模块.
  • 没有必要对现成的共聚焦显微镜进行物理修改.

主要成果:

  • 基于TIE的QPI的可行性在微镜阵列上成功地被证明.
  • 用活植物细胞 () 和哺乳动物培养细胞进行相位成像验证.
  • 实现了结合光和相位信息的多式成像.

结论:

  • 量相成像 (QPI) 可以通过使用TIE技术的商用共聚焦显微镜实现.
  • 这种方法提供了一种非侵入性的方法来测量细胞中的干质量,粘度和温度等物理性质.
  • 未来的应用包括通过相位波动补偿进行偏差校正和散射取消.