超光譜顯微術利用不同物質、組織有不同放射光譜的特性，最開始用於對地球表面上的觀測，後陸續衍伸到多種領域，並同時在生醫醫學上大量應用。其中，擁有光譜高解析度的為空間掃描的系統，又可分為點掃描及線掃描這兩種掃描架構。點掃描普遍的缺點為取樣時間過久，線掃描雖然降低了取樣時間，但犧牲了長軸上的解析度。本研究將線掃描超光譜顯微術與擁有光學切片能力的結構照明顯微術結合，利用結構照明提升長軸上的解析度，成功架設了光學切片超光譜顯微系統。
本研究由光學切片超光譜顯微系統取得目標樣本的螢光超光譜影像，並經由取得的一維光譜影像及二維空間資訊，加上線性分析法的應用，試圖解決光譜量測分析中常見的光譜串擾現象。實驗上使用的樣本有日日春的花粉及多重染色的老鼠毛囊細胞切片，經由實驗結果證明系統的確可以藉由結構照明提升影像的解析度，解決光譜串擾引發的問題，並有效分析出樣本的組成成分。

Hyperspectral imaging makes use of the intrinsic spectral property, in which different materials and tissues solely possess their own emission spectrum. It was originally utilized in remote sensing, and was then developed for a wide variety of fields, including the biomedical field. In hyperspectral imaging systems, high spectral resolution can be achieved by spatial scanning methods which can be further divided into line-scanning approaches and point-scanning approaches. The common disadvantage of the point-scanning approach is its rather long acquisition time, whereas was reduced in line-scanning approaches, but it came with the price of sacrificing its spatial resolution in its long-axial. In this research, line-scanning hyperspectral imaging system is combined with structured illumination to improve the spatial resolution, successfully constructing a hyperspectral imaging system via optical sectioning microscopy.
In this thesis, the hyperspectral imaging system was applied to acquire the fluorescence hyperspectral data of specimens. Utilizing the one-dimensional spectral and two-dimensional spatial information, in addition with the linear unmixing process, the aim is to solve the common crosstalk problem in spectral imaging. With specimens of pollens and multiple stained mouse hair follicles, the imaging results demonstrated that the system can effectively overcome the crosstalk problem, and successfully analyze the constituents of the specimen.

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