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Phase Contrast and Differential Interference Contrast Microscopy01:26

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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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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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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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在斜面平面显微镜中反射直线检测.

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    在Epi-Oblique平面显微镜 (RIDE-OPM) 中反射线内检测简化了高速,高分辨率的生物成像. 这种具有成本效益的系统减少了漂移,并使用标准目标用于先进的,无监督的长期显微镜应用.

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    科学领域:

    • 显微镜的使用方法
    • 生物技术是生物技术.
    • 光学成像技术的成像

    背景情况:

    • 斜照明选择平面显微镜 (EOPM) 能够通过简单的样本安装实现高分辨率,高速成像.
    • 目前的EOPM系统是复杂的,昂贵的,并且由于多个显微镜和专业目标而容易漂移.

    研究的目的:

    • 引入反射直线检测在直角平面显微镜 (RIDE-OPM) 作为一个简化,成本效益的替代方案.
    • 消除了对第三级显微镜和EOPM系统中专门目标的需求.

    主要方法:

    • 开发了一个新的RIDE-OPM系统,集成照明和检测.
    • 使用标准镜头,利用它们的最大数值光圈 (NA) 独立于倾斜角度.
    • 设计了一个紧而稳定的平台,用于无监督的长期成像.

    主要成果:

    • 与传统的EOPM相比,RIDE-OPM显著降低了系统的复杂性和成本.
    • 在视野中实现最佳照明,而不会影响成像速度或质量.
    • 证明了强大的,无监督的长期成像能力,最小的漂移.

    结论:

    • RIDE-OPM为先进的生物成像提供了一种简化,成本效益和稳定的解决方案.
    • 该系统适用于各种生物标本的长期高分辨率成像.
    • 验证了各种样本的性能,包括C. elegans,Drosophila大脑和活细胞.