Characterization and Identification of Fungal Conidia via Shifted Excitation Raman Difference Spectroscopy

Zehua Han, B. Strycker, Blake Commer, Kai Wang, Brain D. Shaw, M. Scully, A. Sokolov
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Abstract

Fungi can be found everywhere, and their impacts on human beings are numerous and varied. Fungi have been widely used in the food, biofuel, beverage and pharmaceutical industries. 1 However, uncontrolled fungal growth can be costly to agriculture, forestry and livestock. If they feed on humans, diseases can be induced such as ringworm, athlete’s foot and lung infections. Some effects on human health may last over years and even lifetimes. 2 It requires timely and accurate identification of mold species to evaluate and/or prevent damage from mold growth, and minimize the consequences of mold exposure. Raman spectroscopy studies on mold spores have been proposed and implemented as a method to identify fungal species. However, the presence of fluorescence emission always hinders Raman signal detection and is virtually impossible to avoid, especially in biological specimens. Shifted excitation Raman difference spectroscopy (SERDS) is a very powerful technique to separate Raman contribution from fluorescence contribution. Herein, we adopt the SERDS modality to extract pure Raman signals from fungal conidia of different species and find that Raman signatures of spores generated from pigment molecules bounded within the cell walls. 3 A further study of conidia of different mold species indicates that the major features of the Raman spectrum correlate with the melanin biosynthesis pathway: species that produce the same melanin exhibit similar Raman spectra. 4 , 5
用位移激发拉曼差分光谱法鉴定真菌分生孢子
真菌无处不在,它们对人类的影响是多种多样的。真菌在食品、生物燃料、饮料和制药工业中有着广泛的应用。然而,不受控制的真菌生长对农业、林业和畜牧业来说代价高昂。如果它们以人类为食,就会引发癣、脚癣和肺部感染等疾病。对人类健康的一些影响可能持续数年甚至一生。它需要及时准确地识别霉菌种类,以评估和/或防止霉菌生长造成的损害,并尽量减少霉菌暴露的后果。真菌孢子的拉曼光谱研究已被提出并应用于真菌种类的鉴定。然而,荧光发射的存在总是阻碍拉曼信号的检测,并且几乎不可能避免,特别是在生物标本中。位移激发拉曼差分光谱(SERDS)是分离荧光和拉曼贡献的一种非常有效的技术。本文采用SERDS方法提取了不同种类真菌分生孢子的纯拉曼信号,发现孢子的拉曼信号是由细胞壁内的色素分子产生的。对不同霉菌物种分生孢子的进一步研究表明,拉曼光谱的主要特征与黑色素生物合成途径相关:产生相同黑色素的物种表现出相似的拉曼光谱。4,5
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