光纤荧光海藻叶绿互a浓度实时测量系统的研究
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摘要
光纤传感技术是海洋测量中应用广泛的重要技术之一。海藻是主要的海洋浮游植物，叶绿素a是海藻中的主要色素。因此，叶绿素a是水生态系统的一个重要参数，其含量是反映海水中浮游植物生物量或现存量的一项重要观测项目，也是计算初级生产力的基础。本系统在利用原有的浓度检测技术前提下，应用光纤作为传输传感元件并结合荧光技术，可以实现海藻浓度高精度的在线测量。本文的主要研究内容包括：
(1)论述荧光测量法的基本原理及特点，对海藻叶绿素a的荧光特性进行分析，为激发光源、滤光片、光探测器的合理选择及使用奠定理论基础。
(2)设计荧光光纤测量系统的总体方案。根据光纤探头的结构、特点，建立荧光信号测量及传输的数学模型，为系统标定提供理论依据。
(3)采用He-Ne激光器作为激发光源，低损耗石英光纤束作为光信号传输通道，高灵敏度、低噪声光电倍增管作为荧光信号探测器，以及以光子计数器为核心的微弱信号检测电路，实现微弱荧光信号的激发、收集、传输、探测和处理。
(4)为了消除激发光源波动对测量结果影响，采用双光路双通道测量方式。为了有效分离叶绿素a所产生的荧光和激发光的散射光，采用高阻塞系数的干涉滤光片作为光谱滤光。
(5)对设计的测量系统设进行实验室定性分析研究，并得出实验结果，包括：叶绿素a的吸收和激发荧光的光谱特性谱线，荧光强度变化与海藻浓度变化之间的线性关系曲线，实验证明用荧光光纤法测量海藻浓度的可行性。
Optical-fiber sensor is one of important technique widely used in the field of ocean measuring. Algae is the main ocean phytoplankton and chlorophyll-a is the main pigment of algae. Chlorophyll-a is one of the important parameters of water ecological system. Its content also is an important observation item which reflects phytoplankton's biomass or stock on hand in the sea, and also the basis of calculating the primary productive forces. Basing on old consistency measuring skill, the system applies optical-fiber as transmitting device with fluorescent technique to realize high efficiency on-line measuring of alga concentration. The main contributions of this dissertation are as follows:
The basic principle and characteristic of fluorescence measurement method are discussed. The fluorescence specific properties of alga chlorophyll-a in the sea are analyzed. Therefore theoretical basis is established for the reasonable selection of excitation light source , optical fiber and optical detector.
The total scheme of fluorescent optical-fiber measuring system is designed. According to the structure and characteristic of optical-fiber probe, the mathematical model of the fluorescent measurement and transfer is established in order to provide theoretical foundation of system calibration.
The system adopts weak signal measuring circuit with He-Ne laser as light house, low-loss silicon optical-fiber bundle as transmission channel of light signal, high sensitivity and low-noise PMT as fluorescent signal detector and photo counter as main detecting device to realize the excitation, collection, transmission, detection and processing of weak fluorescence signal.
To eliminate the signal variation due to undulation of excited light source, the detecting method of double light routes and double channels at the same time are used. In order to effectively separate the fluorescence produced by algae chlorophyll-a and the scatting light from the excitation light, the interference filter with high block coefficient and narrow band is used as spectral filter.
Qualitative analysis and research of the measuring system are processed in the lab to educe the results of experiments, including the absorbing and excited fluorescence spectra character of chlorophyll-a experiments. The linear curve between the variation of fluorescence density and variation of algae concentration is educed and validates the feasibility of the system.
(1)论述荧光测量法的基本原理及特点，对海藻叶绿素a的荧光特性进行分析，为激发光源、滤光片、光探测器的合理选择及使用奠定理论基础。
(2)设计荧光光纤测量系统的总体方案。根据光纤探头的结构、特点，建立荧光信号测量及传输的数学模型，为系统标定提供理论依据。
(3)采用He-Ne激光器作为激发光源，低损耗石英光纤束作为光信号传输通道，高灵敏度、低噪声光电倍增管作为荧光信号探测器，以及以光子计数器为核心的微弱信号检测电路，实现微弱荧光信号的激发、收集、传输、探测和处理。
(4)为了消除激发光源波动对测量结果影响，采用双光路双通道测量方式。为了有效分离叶绿素a所产生的荧光和激发光的散射光，采用高阻塞系数的干涉滤光片作为光谱滤光。
(5)对设计的测量系统设进行实验室定性分析研究，并得出实验结果，包括：叶绿素a的吸收和激发荧光的光谱特性谱线，荧光强度变化与海藻浓度变化之间的线性关系曲线，实验证明用荧光光纤法测量海藻浓度的可行性。
Optical-fiber sensor is one of important technique widely used in the field of ocean measuring. Algae is the main ocean phytoplankton and chlorophyll-a is the main pigment of algae. Chlorophyll-a is one of the important parameters of water ecological system. Its content also is an important observation item which reflects phytoplankton's biomass or stock on hand in the sea, and also the basis of calculating the primary productive forces. Basing on old consistency measuring skill, the system applies optical-fiber as transmitting device with fluorescent technique to realize high efficiency on-line measuring of alga concentration. The main contributions of this dissertation are as follows:
The basic principle and characteristic of fluorescence measurement method are discussed. The fluorescence specific properties of alga chlorophyll-a in the sea are analyzed. Therefore theoretical basis is established for the reasonable selection of excitation light source , optical fiber and optical detector.
The total scheme of fluorescent optical-fiber measuring system is designed. According to the structure and characteristic of optical-fiber probe, the mathematical model of the fluorescent measurement and transfer is established in order to provide theoretical foundation of system calibration.
The system adopts weak signal measuring circuit with He-Ne laser as light house, low-loss silicon optical-fiber bundle as transmission channel of light signal, high sensitivity and low-noise PMT as fluorescent signal detector and photo counter as main detecting device to realize the excitation, collection, transmission, detection and processing of weak fluorescence signal.
To eliminate the signal variation due to undulation of excited light source, the detecting method of double light routes and double channels at the same time are used. In order to effectively separate the fluorescence produced by algae chlorophyll-a and the scatting light from the excitation light, the interference filter with high block coefficient and narrow band is used as spectral filter.
Qualitative analysis and research of the measuring system are processed in the lab to educe the results of experiments, including the absorbing and excited fluorescence spectra character of chlorophyll-a experiments. The linear curve between the variation of fluorescence density and variation of algae concentration is educed and validates the feasibility of the system.
引文
1 [加拿大]T．R．帕森斯C．M．兰莉，[日本]前田吉明．海水分析的化学和生物学方法，厦门：厦门大学出版社，1991．6：5-10
2 Katima Allali, Annick Bricaud, Herve Claustre. Spatial Variability in the Chlorophyll-Specific Absorption Coefficients of Phytoplankton and Photosynthetic Pigment in the Equatorial Pacific. SPIE, 1997,(2963): 179-184
3 Bernhard Ruth. Monitoring the Herbicide Concentration in Ponds by Detecting the Chlorophyll Fluorescence of Algae with Ms-Laser-Excitation. SPIE. 1995,(2503):2-13
4 John J.Cullen, Aurea M.Ciotti, Richard F. Davis. The Relationship between Near-Surface Chlorophyll and Solar-stimulated fluorescence:biological effects. SPIE, 1997,(2693):272-277
5 Robert K.Y. Chan, Ce Chen, S.K.Cheng. Real-time Optical Fiber Sensing of Phytoplankton for Studies in Size Distribution, Concentration and Biomass. SPIE, 1995,(2508): 49-57
6 孙振东，赵玉春，万峰，欧国荣．调制式光纤理化检测仪的研制．仪器仪表学报，2000．4，21(2)
7 喻弼群．藻类对水质的影响及其防治．科技绿野，1997：33-34
8 吴东，王汝霖，陈卫标等．测定叶绿素a浓度、漫散射衰减系数和离水辐亮度的海洋激光雷达．光学学报，1998，18(2)：1690-1696
9 C.M.Lalli, T.R.Parsons. Biological Oceangraphy: An Introduction. Prgamon Press, 1993: 45-50
10 夏达英，王振先，张世魁．荧光技术在海洋环境学上的应用研究．海洋学报，1999，121(3)
11 夏达英，王振先，夏荣环，辛海英．水中荧光计及其在海洋现场探测中的应用．黄渤海海洋，1997，115(2)
12 夏达英．国外荧光技术在海洋学上的应用研究状况．海岸工程．1997，16(1)
13 B.Ruth. A device for the determination of the microsecond component of the invivo chlorophyll fluorescence induction kinetics. Meas. Sci.Technol, 1990,(1):517-521
14 Bernhard Ruth. Monitoring the Herbicide Concentration in Ponds by Detecting the Chlorophyll Fluorescence of Algae with ms-Laser-Excitation. SPIE. 1995, (2503):2-13
15 潘德炉．利用荧光高度遥感海洋中叶绿素 a 的浓度．海洋学报．1989，11(6)：780-787
16 王荣，鲁北纬，余家栋．海水中叶绿素a含量的连续走航测定．海洋与湖沼．1997，28(4)：433-439
17 张季熊，李郁章，王海波．荧光光纤传感器．仪表技术与传感器，1998，6
18 杨性愉．光学检测原理与技术．呼和浩特：内蒙古大学出版社，1994：178-195
19 王荣等．海水叶绿素 a 含量连续走航测定．海洋与湖泊．1996，27：296-301
20 陈国珍，荧光分析，北京：科技出版社，1975：12-25
21 陈卫标，吴东，张亭禄等．海洋表层叶绿素浓度的激光雷达测量方法和海上实验．海洋与湖沼．1998，29(3)：255-260
22 M.L.Paseu, N.Moise, S.Hogiu. Laser Induced Studies on Collagen, Cholesterol and Chlorophyll a. SPIE, 1995,(2527):216-220
23 Raja Holavanahall, Shabbir B.Bambot, Vadde Venkatesh. Noise Measurements of a Phase Fluorometric Instrument. Opt. Eng, 1996, 35(2): 549-553
24 R.Barbini, F. Colao, R.Fantoni, A.Palucci, S.Ribezzo. Fluoresc-ence Determination Pollutants and Natural Components in Seawater from the on Board Mobile Lidar Apparatus. SPIE, 1998, (3534): 359-369
25 H.Barth, R.Heuermann, R.Reuter. In-Situ Analysis of Water Quality Spectral Attenuation and Fluorescence. SPIE, 1997,(3107): 187-194
26 李毛和，张美敦．用于海洋测量的化学和生物光纤传感器．传感器技术学报，2000．3
27 D.Mckee, A.Cunningham, K.Jones. An Integrated Submersible Fluorometer/Nephelometer/Transmissometer: Design and Testing at Sea. Optics & Laser Technology, 1997,29(1):35-39
28 C.J.Lorenzen. A Method for the Continuous Measurement of in vivo Chlorophyll-a Concentration .Deep Sea Res, 1996, (13):223-227
29 傅克忖，曾宪模．海水分光反射率与叶绿素浓度的相关分析．黄渤海海洋，1995，13(1)
30 陈烽．近海机载激光海洋测深技术．应用光学，1999，20(2)：18-23
31 Peter R.C.S.Fearns and Mervyn J.Lynch. Retrieval of chlorophyll concentration via inversion of ocean reflectance—a modeling approach. SPIE, 1997.(2963):408-413
32 George Turner. Review of Fluometric Techniques and Instrum-entation. IEEE Journal of Ocean Engineering, 1979, (5): 583-585
33 C.S.Yentsch and D.A.Phinney. Spedtra Fluorescence: an Ataxonomic Tool for Studying the Structure of Phytoplankton Populations.J.Plank. Res, 1985,7(5):617-632
34 D.S.Knowles,S.H.Lieberman. Field Results from the SCAPS Laser-inducedfluorescence(LIF) Sensor for in-situ Subsurface Detection of Petroleum Hydrocarbons. SPIE, 1995,2504:297-307
35 W. Schade, J.Bublitz. On-site Laser Probe for the Detection of Petroleum Products in Water and Soil. Environ.Sci. Tchnol. 1996,(30): 1451-1458
36 吕海宝．激光光电检测，长沙：国防科技大学出版社，1999．12：1-30
37 张琢．激光干涉测试技术及其应用，北京：机械工业出版社，1998：15-27
38 Hans-Gerd Lohmannsroben, Thomas Roch, Rainer G.Schaefer, Rainer H. Schultze and Harry Vereecken, Laser-Induced Fluorescence (LIF) Spectroscopy for in-situ Analysis of Fluorescence Tracers in water and soils. SPIE, 1997(3107):207-216
39 Vladimir. Haltrin. Chlorophyll-based model of seawater optical properties. Applied optics, 1999,38(33): 6826-6832
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2 Katima Allali, Annick Bricaud, Herve Claustre. Spatial Variability in the Chlorophyll-Specific Absorption Coefficients of Phytoplankton and Photosynthetic Pigment in the Equatorial Pacific. SPIE, 1997,(2963): 179-184
3 Bernhard Ruth. Monitoring the Herbicide Concentration in Ponds by Detecting the Chlorophyll Fluorescence of Algae with Ms-Laser-Excitation. SPIE. 1995,(2503):2-13
4 John J.Cullen, Aurea M.Ciotti, Richard F. Davis. The Relationship between Near-Surface Chlorophyll and Solar-stimulated fluorescence:biological effects. SPIE, 1997,(2693):272-277
5 Robert K.Y. Chan, Ce Chen, S.K.Cheng. Real-time Optical Fiber Sensing of Phytoplankton for Studies in Size Distribution, Concentration and Biomass. SPIE, 1995,(2508): 49-57
6 孙振东，赵玉春，万峰，欧国荣．调制式光纤理化检测仪的研制．仪器仪表学报，2000．4，21(2)
7 喻弼群．藻类对水质的影响及其防治．科技绿野，1997：33-34
8 吴东，王汝霖，陈卫标等．测定叶绿素a浓度、漫散射衰减系数和离水辐亮度的海洋激光雷达．光学学报，1998，18(2)：1690-1696
9 C.M.Lalli, T.R.Parsons. Biological Oceangraphy: An Introduction. Prgamon Press, 1993: 45-50
10 夏达英，王振先，张世魁．荧光技术在海洋环境学上的应用研究．海洋学报，1999，121(3)
11 夏达英，王振先，夏荣环，辛海英．水中荧光计及其在海洋现场探测中的应用．黄渤海海洋，1997，115(2)
12 夏达英．国外荧光技术在海洋学上的应用研究状况．海岸工程．1997，16(1)
13 B.Ruth. A device for the determination of the microsecond component of the invivo chlorophyll fluorescence induction kinetics. Meas. Sci.Technol, 1990,(1):517-521
14 Bernhard Ruth. Monitoring the Herbicide Concentration in Ponds by Detecting the Chlorophyll Fluorescence of Algae with ms-Laser-Excitation. SPIE. 1995, (2503):2-13
15 潘德炉．利用荧光高度遥感海洋中叶绿素 a 的浓度．海洋学报．1989，11(6)：780-787
16 王荣，鲁北纬，余家栋．海水中叶绿素a含量的连续走航测定．海洋与湖沼．1997，28(4)：433-439
17 张季熊，李郁章，王海波．荧光光纤传感器．仪表技术与传感器，1998，6
18 杨性愉．光学检测原理与技术．呼和浩特：内蒙古大学出版社，1994：178-195
19 王荣等．海水叶绿素 a 含量连续走航测定．海洋与湖泊．1996，27：296-301
20 陈国珍，荧光分析，北京：科技出版社，1975：12-25
21 陈卫标，吴东，张亭禄等．海洋表层叶绿素浓度的激光雷达测量方法和海上实验．海洋与湖沼．1998，29(3)：255-260
22 M.L.Paseu, N.Moise, S.Hogiu. Laser Induced Studies on Collagen, Cholesterol and Chlorophyll a. SPIE, 1995,(2527):216-220
23 Raja Holavanahall, Shabbir B.Bambot, Vadde Venkatesh. Noise Measurements of a Phase Fluorometric Instrument. Opt. Eng, 1996, 35(2): 549-553
24 R.Barbini, F. Colao, R.Fantoni, A.Palucci, S.Ribezzo. Fluoresc-ence Determination Pollutants and Natural Components in Seawater from the on Board Mobile Lidar Apparatus. SPIE, 1998, (3534): 359-369
25 H.Barth, R.Heuermann, R.Reuter. In-Situ Analysis of Water Quality Spectral Attenuation and Fluorescence. SPIE, 1997,(3107): 187-194
26 李毛和，张美敦．用于海洋测量的化学和生物光纤传感器．传感器技术学报，2000．3
27 D.Mckee, A.Cunningham, K.Jones. An Integrated Submersible Fluorometer/Nephelometer/Transmissometer: Design and Testing at Sea. Optics & Laser Technology, 1997,29(1):35-39
28 C.J.Lorenzen. A Method for the Continuous Measurement of in vivo Chlorophyll-a Concentration .Deep Sea Res, 1996, (13):223-227
29 傅克忖，曾宪模．海水分光反射率与叶绿素浓度的相关分析．黄渤海海洋，1995，13(1)
30 陈烽．近海机载激光海洋测深技术．应用光学，1999，20(2)：18-23
31 Peter R.C.S.Fearns and Mervyn J.Lynch. Retrieval of chlorophyll concentration via inversion of ocean reflectance—a modeling approach. SPIE, 1997.(2963):408-413
32 George Turner. Review of Fluometric Techniques and Instrum-entation. IEEE Journal of Ocean Engineering, 1979, (5): 583-585
33 C.S.Yentsch and D.A.Phinney. Spedtra Fluorescence: an Ataxonomic Tool for Studying the Structure of Phytoplankton Populations.J.Plank. Res, 1985,7(5):617-632
34 D.S.Knowles,S.H.Lieberman. Field Results from the SCAPS Laser-inducedfluorescence(LIF) Sensor for in-situ Subsurface Detection of Petroleum Hydrocarbons. SPIE, 1995,2504:297-307
35 W. Schade, J.Bublitz. On-site Laser Probe for the Detection of Petroleum Products in Water and Soil. Environ.Sci. Tchnol. 1996,(30): 1451-1458
36 吕海宝．激光光电检测，长沙：国防科技大学出版社，1999．12：1-30
37 张琢．激光干涉测试技术及其应用，北京：机械工业出版社，1998：15-27
38 Hans-Gerd Lohmannsroben, Thomas Roch, Rainer G.Schaefer, Rainer H. Schultze and Harry Vereecken, Laser-Induced Fluorescence (LIF) Spectroscopy for in-situ Analysis of Fluorescence Tracers in water and soils. SPIE, 1997(3107):207-216
39 Vladimir. Haltrin. Chlorophyll-based model of seawater optical properties. Applied optics, 1999,38(33): 6826-6832
40 郭德济主编，光谱分析法(第二版)，重庆：重庆大学出版社，1994．12：162-164
41 廖延彪．光纤光学．北京：清华大学出版社，2000．3：1-16
42 He. Mincai.al. Intramodulated Photodetector. Sensors and Actuator, 1992, A.34:15-19
43 Yensch. C.M, Yentsch. C,S.Emergence of Optical Instrument for Measuring Biological Properties. OceanogMar Biol Ann Rev, 1984,55-98
44 Smith. R.C.Booth, C.R.Star. J.L. Oceanographic Biooptical Profiling System. App Opt, 1984,(23):2791-2797
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