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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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Lens-free Video Microscopy for the Dynamic and Quantitative Analysis of Adherent Cell Culture
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使用一次性定量相梯度成像,以高时间分辨率捕捉细胞形态动态.

Sun Woong Hur1,2, Minsung Kwon1,2, Revathi Manoharaan1,2

  • 1University of Illinois at Urbana-Champaign, Department of Bioengineering, Urbana, Illinois, United States.

Journal of biomedical optics
|July 17, 2024
PubMed
概括

快速,无标签的定量相成像 (ss-QPGM) 实时监测细胞变化. 这种技术揭示了细胞死亡期间的形态和生化变化,为活细胞动态提供了一个有前途的工具.

关键词:
细胞死亡是细胞死亡.没有标签的成像技术.形态学 形态学 形态学定量阶段成像成像技术的使用.一次射击成像成像技术

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科学领域:

  • 生物医学光学 生物医学光学
  • 细胞生物学 细胞生物学
  • 显微镜的使用方法

背景情况:

  • 无标签量化相位成像 (QPI) 提供了对细胞动态的不扰乱测量.
  • 开发更快,更灵敏的仪器对于捕捉快速细胞事件至关重要.
  • 单拍定量相梯度显微镜 (ss-QPGM) 能够同时获取极化元件以进行相位重建.

研究的目的:

  • 描述一种快速的,一次性定量相梯度显微镜 (ss-QPGM) 方法.
  • 为了记录单细胞相位图像的形态变化.
  • 使用光成像,将这些形态变化与细胞死亡的生化指标相关联.

主要方法:

  • 实现了一个计算高效的最小平方算法,用于实时,视频速率成像 (高达30 fps).
  • 利用ss-QPGM获取相位图像,并同时获取生物化学标记细胞的光图像.
  • 使用USAF1951模式的相位目标和校准的样本厚度验证了相位测量准确性.

主要成果:

  • 实现了0.8微米的空间分辨率和精确的相位检索 (R2>0.99).
  • 实时显示,视频速率 (30 fps) ss-QPGM用于活细胞成像.
  • 在营养缺乏和药物诱导的细胞死亡期间观察到动态的细胞内和形态变化,与死亡相关.

结论:

  • 无标签的ss-QPGM为活细胞成像提供了高时间分辨率和高空间精度.
  • 该技术有效地监测诱导细胞死亡期间的动态细胞变化.
  • ss-QPGM显示出对研究活细胞动态与最小的干扰显著的前景.