光谱学与光谱分析, 2019, 39 (3): 802, 网络出版: 2019-03-19  

小型荧光激光雷达测量六种典型生物战剂模拟物二维荧光光谱

2D Fluorescence Spectra Measurement of 6 Kinds of Bioagents Simulants by Short-Range Lidar
作者单位
1 中国人民解放军陆军炮兵防空兵学院, 安徽 合肥 230031
2 安徽农业大学生命科学学院, 安徽 合肥 230031
3 中国科学院安徽光学精密机械研究所, 安徽 合肥 230031
摘要
对成团泛菌(Pan)、 金黄色葡萄球菌金黄亚种(Sta)、 球芽孢杆菌(BG)和大肠杆菌(EH)四种生物战剂模拟物进行了培养和生长曲线测定, 四种菌的代时分别为0.99, 0.835, 1.07和1.909 h; 设计研制了近场小型荧光测量激光雷达, 266和355 nm波长分别用于测量生物战剂模拟物氨基酸段和NADH段二维荧光谱; 在可控的荧光测量腔室, 测得了分辨率为4 nm的营养态细菌液态气溶胶以及牛血清(BSA)、 卵清蛋白(OVA)两种毒素类模拟物液态气溶胶的二维荧光谱; 二维荧光谱数据表明, Pan, Sta, BG, EH, BSA和OVA气溶胶, 在氨基酸段的荧光谱形与标准荧光组分色氨酸较为一致, FWHM为60 nm, 受培养生化环境、 细菌内部荧光组分及比例的影响, 荧光分子激发态与基态间的能量差增大, 荧光谱带均存在不同程度的蓝/紫移; 在Pan, Sta, BG和EH营养细菌气溶胶中均检出了较弱的NADH荧光组分, 且水、 氮等的拉曼散射不能完全扣除, 光谱锯齿严重, FWHM为100 nm; 二阶求导后的二维荧光谱表明, 荧光谱的高阶处理和分辨识别是可行的。
Abstract
Four kinds of bioagents simulants, Pantoea agglomerans (Pan), Staphylococcus aureus subsp. Aureus (Sta), Bacillus globigii (BG) and Escherichia coli (EH) were cultured and the strains growth curves were obtained, and the generation time were 0.99, 0.835, 1.07 and 1.909 h respectively. Two wavelengths at 266 and 355 nm of a designed short range fluorescence lidar were engaged for the measurements of two dimensional fluorescence spectra in the amino acid band and NADH band of the biological warfare agents, respectively. The simulants were diffused homogeneously inside a controlled fluorescence measurement chamber and interogated by the lidar. The 2 dimensional fluorescence spectra of four kinds of vegetative bacteria, BSA and OVA(as simulant of toxin agents) were obtained with resolution of 4 nm. The fluorescence spectra of Pan, Sta, EH and BG, BSA and OVA were consistent with the standard fluorescent component tryptophan in the amino acid band with FWHM of 60 nm, but the central wavelength of the fluorescence spectra of these simulants blue/purple shifted obviously as affected by the external biochemical environment, concentration and ratio of different bacterial internal fluorophores, so the energy level between the excited state and the ground state of the fluorescence molecule increased. Accordingly, weak NADH fluorescence spectra with 100 nm FWHM inside the four vegetative bacteria aerosols were also detected, but Raman scattering contribution of water and nitrogen could not be effectively extracted. The second-order derivative fluorescence spectra of the four simulants showed that the high-order processing and recognition of the fluorescence spectrum was feasible.

杨辉, 马修兵, 孙彦飞, 王铁栋, 庆丰, 赵雪松. 小型荧光激光雷达测量六种典型生物战剂模拟物二维荧光光谱[J]. 光谱学与光谱分析, 2019, 39(3): 802. YANG Hui, MA Xiu-bing, SUN Yan-fei, WANG Tie-dong, QING Feng, ZHAO Xue-song. 2D Fluorescence Spectra Measurement of 6 Kinds of Bioagents Simulants by Short-Range Lidar[J]. Spectroscopy and Spectral Analysis, 2019, 39(3): 802.

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