Fluorescence Spectroscopy and Microscopy
Fluorescence Spectroscopy and Microscopy
复制标题
荧光光谱和显微镜
作者：
Y. Engelborghs;A. Visser
当代的荧光分光光度计包括激发光源、激发和发射单色器以及检测器，其在没有校正的情况下产生不符合理想光谱响应的数据。激发光路和发射光路的光谱特性的校正首先需要波长和光谱精度的校准。激发光路可以使用光谱校正的参考检测系统校正到样品位置。校正的参考响应考虑了激发光源相对于发射和/或透射检测器响应的光谱强度和漂移。发射检测路径还必须针对样品室光学器件、发射单色器和检测器的组合光谱偏差进行校正。存在与激发和发射校正两者相关联的几个关键问题，包括考虑光谱带通和分辨率、光学带通或中性密度滤光片以及光路中偏振元件的位置和方向的要求。此外，还描述了二次校正因子，包括（1）减去溶剂的荧光背景，（2）去除Rayleigh和拉曼散射线，以及（3）校正与样品浓度相关的内滤光片效应。美国国家标准与技术研究所（NIST）可追溯的校准和校正协议的重要性解释在有效的实验室内和实验室间的研究和有效的光谱定性和定量分析，包括多元光谱建模。
Contemporary spectrofluorimeters comprise exciting light sources, excitation and emission monochromators, and detectors that without correction yield data not conforming to an ideal spectral response. The correction of the spectral properties of the exciting and emission light paths first requires calibration of the wavelength and spectral accuracy. The exciting beam path can be corrected up to the sample position using a spectrally corrected reference detection system. The corrected reference response accounts for both the spectral intensity and drift of the exciting light source relative to emission and/or transmission detector responses. The emission detection path must also be corrected for the combined spectral bias of the sample compartment optics, emission monochromator, and detector. There are several crucial issues associated with both excitation and emission correction including the requirement to account for spectral band-pass and resolution, optical band-pass or neutral density filters, and the position and direction of polarizing elements in the light paths. In addition, secondary correction factors are described including (1) subtraction of the solvent’s fluorescence background, (2) removal of Rayleigh and Raman scattering lines, as well as (3) correcting for sample concentration-dependent inner-filter effects. The importance of the National Institute of Standards and Technology (NIST) traceable calibration and correction protocols is explained in light of valid intraand interlaboratory studies and effective spectral qualitative and quantitative analyses including multivariate spectral modeling.
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影响因子：
2
作者：
GROSSWEINER, LI
通讯作者：
GROSSWEINER, LI
影响因子：
--
作者：
M. L. Johnson;L. M. Faunt
通讯作者：
M. L. Johnson;L. M. Faunt
亚硝酸盐/α晶状体蛋白反应：晶状体基质损伤的可能机制。
DOI：
10.1006/exer.1999.0761
发表时间：
2000
期刊：
Experimental eye research.
影响因子：
--
作者：
Paik,DC;Dillon,J
通讯作者：
Dillon,J
影响因子：
62.1
作者：
Jameson, David M.;Ross, Justin A.
通讯作者：
Ross, Justin A.
影响因子：
3.5
作者：
Gratton, E;Breusegem, S;Ruan, QQ
通讯作者：
Ruan, QQ