Microspectroscopic Studies on Sample Dimension and Dye Distribution Characteristics.

给出了不同形状和大小(样品尺寸；D)的小体积中分析物(即染料)的光谱数据(吸光度、荧光强度或荧光寿命)的模型和相关方程，并讨论了影响数据的因素。光谱性质，特别是染料的吸光度，被证明依赖于三维样品结构：薄膜(1-D)、管(2-D)和球体(3-D)。尺寸效应对吸光度的影响大小由样品和探针束的大小决定。此外，染料在2-D和3-D体系中的分布特性也影响吸光度；仅分布在2-D或3-D样品表层的染料的吸光度分别是该均匀分布的0.50或0.33倍。如果一种染料单独分布在样品的内部体积中，则吸光度比均匀分布时的吸光度增加。在所提出的模型的基础上，讨论了样品尺寸、染料分布特性和探针束尺寸对吸收显微光谱的影响。同样，样品尺寸以及染料的分布特性也会影响荧光强度和荧光寿命。对于液体和聚合物膜(1-D)、微毛细管、微通道芯片和液/液界面(2-D)以及液滴、聚合物小球、微胶囊、巨泡(3-D)等样品，详细讨论了样品尺寸和染料分布特性对光谱数据的影响。

Models and the relevant equations giving spectroscopic data (optical absorbance, fluorescence intensity, or fluorescence lifetime) of an analyte (I.e., dye) confined in a small volume with different shapes and sizes (sample dimension; D) were given and, factors governing the data were discussed. The spectroscopic properties, particularly dye absorbance, were shown to be dependent on the three-dimensional sample structure: films (1-D), tubes (2-D), and spheres (3-D). The magnitude of the dimension effect on absorbance was determined by the size of both sample and probe beam. Furthermore, dye distribution characteristics in 2-D and 3-D systems also affected absorbance; a dye distributed exclusively to the surface layer of a 2-D or 3-D sample gave an absorbance 0.50 or 0.33-fold of that homogeneously distribution, respectively. If a dye distributed in the inner volume of the sample alone, absorbance increases over that for homogeneous distribution. Effects of sample dimension, dye distribution characteristics, and the size of the probe beam in absorption microspectroscopy were discussed on the basis of the proposed models. Similarly, the sample dimension as well as the dye distribution characteristics affected fluorescence intensity and fluorescence lifetime. For samples such as liquid and polymer films (1-D), microcapillary, microchannel chips, and liquid/liquid interface (2-D), and liquid droplets, polymer beads, microcapsules, giant vesicles (3-D), effects of sample dimension and dye distribution characteristics on the spectroscopic data were discussed in detail.