氨酸染料的氨酸灭作为光显微镜的多功能工具
Joshua C Vaughan1, Graham T Dempsey, Eileen Sun
1Howard Hughes Medical Institute, and Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Journal of the American Chemical Society
|January 15, 2013
概括
某些色素染料,如Cy5,可以通过三2-碳氧乙烯) 素 (TCEP) 进行可逆灭. 这种素相互作用允许在超高分辨率成像和细胞分子分化中的应用.
科学领域:
- 化学生物学 化学生物学
- 生物物理化学 生物物理化学
- 分子成像学分子成像学
背景情况:
- 氨酸染料是广泛使用的光探针.
- 可逆火机制对于先进的成像技术非常有价值.
研究的目的:
- 通过三二碳氧乙 (TCEP) 来研究氨酸染料Cy5的灭.
- 探索TCEP介导火的机制及其潜在应用.
主要方法:
- 对Cy5-TCEP相互作用的光谱分析.
- 摄影化学研究涉及紫外线光诱导的 adduct 分解.
- 在超分辨率成像中演示TCEP火.
- 在区分细胞内和细胞外分子中的应用.
主要成果:
- TCEP可逆地灭Cy5和相关的氨酸染料的光.
- 火是通过TCEP的1,4-添加到染料的聚甲桥,形成一个共价添加物发生的.
- 紫外线照明会使附加物解离,恢复光.
- TCEP火使得超高分辨率成像和细胞定位研究成为可能.
结论:
- TCEP是一种有效的可逆灭剂,用于像Cy5.5这样的氨酸染料.
- 该机制涉及可逆的共价 adduct 形成.
- 这种可逆火为先进的光学成像和生物分析提供了新的机会.
相关概念视频
Photoluminescence: Applications
Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
Fluorescence and Phosphorescence: Instrumentation
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.
Variables Affecting Phosphorescence and Fluorescence
Fluorescence and phosphorescence are essential phenomena in fields like analytical chemistry, biological imaging, and materials science, where they detect molecular properties and visualize cellular structures. Understanding the variables that influence these luminescent behaviors is crucial for maximizing accuracy and efficiency in their applications. These variables can broadly be grouped into chemical structure, solvent properties, and external conditions, each playing a distinct role in...
Photoluminescence: Fluorescence and Phosphorescence
Photoluminescence is a process where a molecule absorbs light energy and re-emits it in the form of light. This phenomenon occurs when a substance absorbs photons, promoting its electrons to higher energy level excited states, followed by a relaxation process in which the electrons return to their original ground state energy levels and emit light. Photoluminescence is widely observed in various materials, including semiconductors, and organic and inorganic compounds.
A pair of electrons in a...
A pair of electrons in a...
Super-resolution Fluorescence Microscopy
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 developed.


