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相关概念视频

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

7.1K
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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Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Fluorescence and Phosphorescence: Instrumentation01:25

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Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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Confocal Fluorescence Microscopy01:16

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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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Conducting Multiple Imaging Modes with One Fluorescence Microscope
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光激发发射矩阵成像

Oren Katz1, Travis Ferguson2, Emma Abbey1

  • 1Department of Chemistry, University of Victoria, Victoria, British Columbia V8W 5C2, Canada.

Analytical chemistry
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此摘要是机器生成的。

这项研究引入了一个快速的4D光成像系统,能够捕捉每个像素的激发-发射-矩阵光谱. 这种新系统能够快速获取数据,用于复杂的光样本分析.

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科学领域:

  • 频谱学是一种光谱学.
  • 显微镜的使用方法
  • 分析化学 分析化学

背景情况:

  • 传统的光成像通常面临着光谱分辨率和采集速度的限制.
  • 描述复杂的光样品需要详细的光谱信息跨空间维度.

研究的目的:

  • 开发和验证一种新的四维 (4D) 光成像系统.
  • 提高每个像素的激发发射矩阵 (EEM) 光谱的数据采集率.
  • 证明系统在分析复杂混合物和动态光变化的能力.

主要方法:

  • 实施一个具有65,536个像素的4D光成像系统,每个像素都包含一个激发发射矩阵光谱 (31个激发,8个发射波长).
  • 采用哈达马德变换多重复合与31通道可编程光源来快速获取数据 (每张图像8秒).
  • 采用多变量分析,特别是并行因子分析,用于数据解释.

主要成果:

  • 成功获取跨空间维度 (x,y,激发波长,辐射波长) 的4D EEM光谱数据.
  • 在不同温度下对毛细血管中的染料溶液和分层溶液进行成像,证明了系统性能.
  • 生成的光体的空间分布图像及其相对强度随时间变化.

结论:

  • 开发的4D光成像系统显著提高了EME光谱的数据采集速度.
  • 该系统有效地表征复杂的光混合物和动态光谱变化.
  • 多变量分析为光体分布和行为提供了有价值的见解.