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相关概念视频

Photoreceptors and Visual Pathways01:22

Photoreceptors and Visual Pathways

5.6K
At the molecular level, visual signals trigger transformations in photopigment molecules, resulting in changes in the photoreceptor cell's membrane potential. The photon's energy level is denoted by its wavelength, with each specific wavelength of visible light associated with a distinct color. The spectral range of visible light, classified as electromagnetic radiation, spans from 380 to 720 nm. Electromagnetic radiation wavelengths exceeding 720 nm fall under the infrared category,...
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Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

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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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相关实验视频

Updated: Jun 3, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
09:57

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

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基于光物理的两光子电压成像使用FRET-opsin电压指示器.

F Phil Brooks1, Daozheng Gong1, Hunter C Davis1

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Science advances
|January 8, 2025
PubMed
概括

基因编码的电压指示器 (GEVIs) 在双光子激发下显示电压敏感性问题. 优化的协议使得使用FRET-opsin GEVIs (如Voltron2.2) 的高速度体内电压成像成为可能.

科学领域:

  • 神经科学是一个神经科学.
  • 生物物理学的生物物理.
  • 分子生物学分子生物学

背景情况:

  • 来自微生物罗多素的遗传编码电压指标 (GEVIs) 对于绘制生物电活动的地图至关重要.
  • 福斯特共振能量转移 (FRET) -opsin GEVI 提供高亮度,速度和电压灵敏度.
  • 一个显著的局限性是FRET-opsin GEVIs在两光子 (2P) 激发下的降低电压灵敏度.

研究的目的:

  • 为了研究FRET-opsin GEVIs Voltron1和Voltron2.2的光物理,
  • 了解2P激发下电压敏感性下降的原因.
  • 为有效的2P电压成像 in vivo开发优化的协议.

主要方法:

  • 沃尔特朗1和沃尔特朗2的光物理特征.
  • 分析电压灵敏度作为照明强度的函数.
  • 开发和应用光循环优化的2P照明协议.
  • 在活体中2P电压成像在一个活的小鼠模型.

主要成果:

  • 在FRET-opsin GEVIs中观察到的电压灵敏度来源于光循环中间体,而不是基态.
  • 电压灵敏度是照明强度的非线性函数,在低强度下可能出现信号反转.

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Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

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In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy
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In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

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Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
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Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

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In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy
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In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

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  • 成功开发了光循环优化的2P照明协议.
  • 高速2P电压成像在体内使用Voltron2在小鼠大脑中进行了证明.
  • 结论:

    • 在2P激发下FRET-opsin GEVIs的光物理是复杂的,受到光循环中间体和照明强度的影响.
    • 优化的2P照明协议可以克服以前的局限性,从而实现有效的体内电压成像.
    • 这些发现为神经科学中先进的高速2P电压成像应用铺平了道路.