A unique computer centered instrument for simultaneous absorbance and fluorescence measurements
A unique computer centered instrument for simultaneous absorbance and fluorescence measurements
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一种独特的以计算机为中心的仪器，可同时测量吸光度和荧光
DOI：
10.1021/ac60323a009
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发表时间：
1973
影响因子：
7.4
通讯作者：
A. Timnick
中科院分区：
文献类型：
--
作者：
J. Holland;R. Teets;A. Timnick
本文介绍了一种以计算机为中心的荧光光谱仪-吸收光谱仪组合装置，它可以同时进行吸收光谱和荧光光谱的测量。专用计算机对收集的荧光测量的许多仪器和物理变量的数据进行校正。从激发扫描过程中收集的数据，计算机可以输出吸光度、荧光、量子校正荧光和唯一的量，即部分量子效率。通过发射扫描，可以获得吸光度、荧光、量子校正荧光和总量子效率。吸收校正荧光是指通过样品池中的吸收过程来校正激发光束衰减的测量方法。详细介绍了计算机程序，并给出了各种吸收和荧光测量的结果，以评价该系统在化学分析中的应用，但迄今为止，对显式观测的充分理解还没有完全达到。准确的荧光模型对于面临缺乏统一标准化、准确性较差以及在许多情况下极端稀释等问题的化学分析员来说尤为重要。在荧光中，测量的量不像分光光度法那样减少到简单的比率，并且存在基本的几何和波长相关因素，这些因素对于每个荧光仪器都是唯一的。尽管荧光分析的基本复杂性，但人们认为，通过更有效地控制测量变量，可以获得更高的准确度和不同仪器测量之间更好的相关性。荧光测量中涉及的因素可分为两类，仪器因素和物理因素。仪器因素与所用设备和观察条件有关，而物理方面涉及在观察期间发生的分子、溶剂笼或溶液内的能量吸收、再发射和竞争性损失。
A computer centered spectrophotometer-spectrofluo-rimeter combination instrument has been fabricated that will allow simultaneous absorption and fluores-cence measurements. The dedicated computer ap-plies corrections to the data collected for many of the instrumental and photophysical variables of fluores-cence measurements. From datacollected during an excitation scan, the computer can output absorbance, fluorescence, quanta corrected fluorescence, and a unique quantity, partial quantum efficiency. From an emission scan, absorbance, fluorescence, quantum corrected fluorescence, and total quantum efficiency can be obtained. Absorption corrected fluorescence is the name proposed for measurements which correct for the attenuation of excitation beam by absorption processes in thesample cell. The computer program is detailed and results are presented to evaluate the system for the various absorption and fluorescence measurements.Applications of fluorescence to chemical analysis have been widespread, but an adequate understanding of the man-ifested observations has not been completely attained to this date. An accurate model for fluorescence is of particular im-portance to the chemical analyst who is faced with the prob-lems of lack of uniform standardization, relatively poor ac-curacy, and in many cases extreme dilutions. In fluorescence, measured quantities do not reduce to a simple ratio as in spectrophotometry and there are basic geometric and wavelength dependent factors which are unique for each fluorescence instrument. In spite of the fundamental complexities of fluorescence analysis, it is felt that greater ac-curacy and better correlation between measurements made with different instruments can be attained with more effective control of the measurement variables. The factors involved in fluorimetric measurement can be divided into two categories, instrumental and photophysical. The instrumental factors pertain to the equipment employed and the conditions of observation, while the photophysical aspects concern the absorption, re-emission, and competitive losses of energy within the molecule, solvent cage, or solution occurring during the period of observation.