海水中浮游植物粒径分布及浓度原位监测系统研究
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摘要
浮游植物是海洋生态系统中最主要的生产者和能量转换者，其过度的增殖会造成海洋环境的恶化和“赤潮”的频发。对浮游植物种群结构、生物量变化等的原位测量可以及时监测海洋生态环境的变化，对“赤潮”等自然灾害起到预警的作用。粒径是表征浮游植物的一个重要参数，浮游植物的新陈代谢、光合作用、能量利用等生理特性都与其粒径大小和分布有关。在对浮游植物现场监测方法和粒径测量方法深入研究的基础上，本文研制了海水中浮游植物浓度及其粒径分布的原位实时自动监测系统，实现了海洋浮游植物粒径分布及种群密度的实时、定点、快速监测。
本文所设计的监测系统采用激光激发荧光与激光差分多普勒技术相结合的方法，由激光激发荧光信号确定浮游植物的存在和经过测量区域的时间宽度，由差分多普勒信号确定粒子的速度，计算得到浮游植物的粒径和浓度。系统由激光器、激光光纤耦合装置、水下光学探头、光电检测和数据处理四部分构成。本文的工作是对系统的四部分进行了具体的设计和调试；进行了系统联调和定标；系统实验室的性能测试和数据分析，现场的原位测量。具体的工作包括：
1.在深入研究基于激光激发荧光和激光差分多普勒技术相结合的浮游植物粒径测量方法的基础上，设计了测量系统的总体结构；选择、确定了激光器、加工了传输光纤；研究了高斯光束的特性和高斯光束单透镜变换特性，设计、调试了激光光纤单透镜耦合装置，对于芯径是3.5m，数值孔径是0.13,长度是100m的单模光纤，耦合效率可达到50%，为光学探头的测量提供了良好的入射光信号。
2.研制了适合于现场原位测量的水下测量装置：水下光学探头。首先设计、调试了水下光学探头的内部光路。在光路设计上，将激发荧光信号的接收方向和激光多普勒信号的接收方向设计成和入射光传输方向成90°角，位于入射光传输方向的两侧，使接收到的激发荧光信号和激光多普勒信号同步，粒径分布的计算更准确；采用横向分束器设计了一鲁棒性较好的分束装置，即使装置安装有轻微的倾斜或海水的涌动使入射光的方向变化也不会影响输出光束的稳定性。其次设计、加工了一密封性良好的光学探头密封舱和探头固定支架，实验室和现场实验表明，光学探头密封舱和固定支架都符合系统的测量要求。
3.设计了水上光电检测电路和基于DSP的水上数据采集装置的硬件，编写了针对于硬件的采集软件和上位机的数据处理、存储和显示软件。
4.对设计的系统进行了联调，形成了测试仪器样机，对系统的可靠性进行了测试，分析了系统的误差，进一步调整了系统。在实验室，用培养的三种形状不同粒径大小的12种浮游植物培养液进行了单一藻种和混合藻种的比测，用高斯函数对粒径分布进行了拟合。将样机和CASY-TT型细胞分析仪的测试结果做了分析对比，给出了系统样机的单一藻种粒径分布误差和浓度误差。粒径范围在5～50m的椭球和球形的藻种，粒径分布统计分布误差在10%以内；细胞分析仪和样机的粒径均值和峰值都有较好的线性相关性；混合藻种的浓度误差在5%左右。
5.在青岛近海进行了2次多个站点的现场原位测量，测量结果表明，系统样机能够测得海水中浮游植物的粒径分布和浓度信息，测量探头可以放入深至14米的水下，仪器调试好后系统可以长时间自动运行。可用于现场原位的连续、实时的监测。从测试数据来看，测试水域的粒子浓度变化不大，和显微镜观察的结果误差在可接受的范围内。
本文所设计、研制的海水中浮游植物浓度和粒径分布原位测量系统，测量的核心部件光学探头完全由光学器件搭建，不需供电，完全密封，可长时间放入水下，由光纤传输测量信号，可实现长时间连续的现场原位监测。和已有其他的浮游植物现场原位测试系统相比，系统结构简单，放置方便，不需要经过复杂算法分析，可直接测得浮游植物单粒子的粒径，系统实时性较好。该系统可望在海洋环境监测与评价、赤潮研究、海洋生态过程研究、水质监测等领域发挥作用。
Phytoplankton is the major producer and energy transducer of marine ecosystems,and its excessive proliferation will worsen the marine environment and lead to more“red tides”. The in situ measurement of phytoplankton population structure andbiomass can be used to monitor the change in marine environment and to predict “redtides” and other natural disasters. As an important parameter, particile size distributionis related to the physiological properties of phytoplankton, such as metabolism,photosynthesis and energy use. In this thesis, an in situ measurement system isdeveloped to achieve real-time and automatic monitoring of phytoplankton sizedistribution and population density in coastal seawaters.
The monitoring system combines laser-induced fluorescence detection and laserdifferential Doppler velocimetry. The former is used to identify the phytoplanktonparticles passing the detection zone, and to determine the time it take for thephytoplankton particles to transverse interference fringes in the laser intersectionvolume, while the latter is used to obtain the velocity parameter of the phytoplanktonparticles passing the detection zone, thus the size and concentration can be deduced.The system consists of a laser, a laser-fiber coupling device, an underwater opticalprobe and a data acquisition&processing part. In the thesis, the designing&debugging of all the whole system, the systematic calibration, laboratory tests, dataanalysis and in situ measurements was described. Specific works done are as follows.
1. By in-depth research on the measuring method, an overall scheme of themeasurement system was determined. A532nm laser was selected; and a laser tosingle-mode fiber coupling device was made on the the basis of the characters ofGaussian beam and its transformation characters through an aspheric lens, which canachieve the coupling efficiency of50%for a100m single-mode fiber with a core diameter of3.5m and a numerical aperture of0.13.
2. An underwater optical probe, a watertight chamber housing the optical sensingsystem, was developed. The laser transmitted by a single-mode fiber entered thechamber, then split into two parallel beams and intersected with each other outside thechamber. The beam intersection defines the measurement volume in the seawater. Thedirections of±90o deviating from the transmission direction of laser beams arerespectively chosen to receive laser-induced fluorescence and laser Doppler signals inorder to keep both signals synchronic to get accurate size distribution. A robustbeam-splitting device, with output intensity insensitive to the incident beam directionchange caused by slight incident inclination and sea waves, was designed, using alateral displacement beamsplitter. Lab and field experiments proved that the opticalprobe chamber with its support system met the measurement requriements.
3. Photoelectric detection circuits and data acquisition&processing hardwareswere designed. DSP programs&PC software units for data processing, storage anddisplay were designed and completed.
4．The measurement system prototype was debugged, tested, further adjusted inthe laboratory, and the error analysis was done. In the laboratory tests and calibrationexperiments,12phytoplankton species in3different kinds of shapes cultured in the labwere used. Gaussian curve-fitting method was adopted to study the phytoplankton sizedistribution data. The results of lab experiments were compared with that measured byCASY-TT cell counter+analyser system. Corresponding size distribution andconcentration errors for all kinds of phytoplankton measured in the tests were given.Statistical distribution errors of size distributions for spherical or ellipticalphytoplankton with a size range of5-50m were less than10%. The average size valuemeasured by the prototye was well linearly relevant with the one measured byCASY-TT, The concentration measurement errors of mixed phytoplankton were around5%.
5. The prototype in situ experiments were done twice at different stations inQingdao coastal waters, and the results proved that the designed system can be used for in situ measurement of size distribution and concentration information of marinephytoplankton, The adjusted system can also do real-time monitoring at a fixedposition for a long time. The experimental data demonstrated no distinct change ofphytoplankton concentration in the measured waters. Compared with microscopesobservation results, the measurement errors are within an acceptable range.
The system developed in the thesis is capable of real-time size distribution andconcentration measurement for phytoplankton particles. The underwater opticalsensing probe does not rely on electrical power, instead, it uses optical fibers totransmit laser and the measured optical signals. The sensing probe can stay submergedfor long period of time, making a long-term real-time in situ monitoring possible. In thesame time, compared with the other measurement methods, the system needs neithercomplicated optics nor complex algorithms, so it is of outstanding real-timeperformance. The system has a promising application in marine environmentmonitoring and evaluation,“red tide” research, marine ecosystem process research andwater quality monitoring.
本文所设计的监测系统采用激光激发荧光与激光差分多普勒技术相结合的方法，由激光激发荧光信号确定浮游植物的存在和经过测量区域的时间宽度，由差分多普勒信号确定粒子的速度，计算得到浮游植物的粒径和浓度。系统由激光器、激光光纤耦合装置、水下光学探头、光电检测和数据处理四部分构成。本文的工作是对系统的四部分进行了具体的设计和调试；进行了系统联调和定标；系统实验室的性能测试和数据分析，现场的原位测量。具体的工作包括：
1.在深入研究基于激光激发荧光和激光差分多普勒技术相结合的浮游植物粒径测量方法的基础上，设计了测量系统的总体结构；选择、确定了激光器、加工了传输光纤；研究了高斯光束的特性和高斯光束单透镜变换特性，设计、调试了激光光纤单透镜耦合装置，对于芯径是3.5m，数值孔径是0.13,长度是100m的单模光纤，耦合效率可达到50%，为光学探头的测量提供了良好的入射光信号。
2.研制了适合于现场原位测量的水下测量装置：水下光学探头。首先设计、调试了水下光学探头的内部光路。在光路设计上，将激发荧光信号的接收方向和激光多普勒信号的接收方向设计成和入射光传输方向成90°角，位于入射光传输方向的两侧，使接收到的激发荧光信号和激光多普勒信号同步，粒径分布的计算更准确；采用横向分束器设计了一鲁棒性较好的分束装置，即使装置安装有轻微的倾斜或海水的涌动使入射光的方向变化也不会影响输出光束的稳定性。其次设计、加工了一密封性良好的光学探头密封舱和探头固定支架，实验室和现场实验表明，光学探头密封舱和固定支架都符合系统的测量要求。
3.设计了水上光电检测电路和基于DSP的水上数据采集装置的硬件，编写了针对于硬件的采集软件和上位机的数据处理、存储和显示软件。
4.对设计的系统进行了联调，形成了测试仪器样机，对系统的可靠性进行了测试，分析了系统的误差，进一步调整了系统。在实验室，用培养的三种形状不同粒径大小的12种浮游植物培养液进行了单一藻种和混合藻种的比测，用高斯函数对粒径分布进行了拟合。将样机和CASY-TT型细胞分析仪的测试结果做了分析对比，给出了系统样机的单一藻种粒径分布误差和浓度误差。粒径范围在5～50m的椭球和球形的藻种，粒径分布统计分布误差在10%以内；细胞分析仪和样机的粒径均值和峰值都有较好的线性相关性；混合藻种的浓度误差在5%左右。
5.在青岛近海进行了2次多个站点的现场原位测量，测量结果表明，系统样机能够测得海水中浮游植物的粒径分布和浓度信息，测量探头可以放入深至14米的水下，仪器调试好后系统可以长时间自动运行。可用于现场原位的连续、实时的监测。从测试数据来看，测试水域的粒子浓度变化不大，和显微镜观察的结果误差在可接受的范围内。
本文所设计、研制的海水中浮游植物浓度和粒径分布原位测量系统，测量的核心部件光学探头完全由光学器件搭建，不需供电，完全密封，可长时间放入水下，由光纤传输测量信号，可实现长时间连续的现场原位监测。和已有其他的浮游植物现场原位测试系统相比，系统结构简单，放置方便，不需要经过复杂算法分析，可直接测得浮游植物单粒子的粒径，系统实时性较好。该系统可望在海洋环境监测与评价、赤潮研究、海洋生态过程研究、水质监测等领域发挥作用。
Phytoplankton is the major producer and energy transducer of marine ecosystems,and its excessive proliferation will worsen the marine environment and lead to more“red tides”. The in situ measurement of phytoplankton population structure andbiomass can be used to monitor the change in marine environment and to predict “redtides” and other natural disasters. As an important parameter, particile size distributionis related to the physiological properties of phytoplankton, such as metabolism,photosynthesis and energy use. In this thesis, an in situ measurement system isdeveloped to achieve real-time and automatic monitoring of phytoplankton sizedistribution and population density in coastal seawaters.
The monitoring system combines laser-induced fluorescence detection and laserdifferential Doppler velocimetry. The former is used to identify the phytoplanktonparticles passing the detection zone, and to determine the time it take for thephytoplankton particles to transverse interference fringes in the laser intersectionvolume, while the latter is used to obtain the velocity parameter of the phytoplanktonparticles passing the detection zone, thus the size and concentration can be deduced.The system consists of a laser, a laser-fiber coupling device, an underwater opticalprobe and a data acquisition&processing part. In the thesis, the designing&debugging of all the whole system, the systematic calibration, laboratory tests, dataanalysis and in situ measurements was described. Specific works done are as follows.
1. By in-depth research on the measuring method, an overall scheme of themeasurement system was determined. A532nm laser was selected; and a laser tosingle-mode fiber coupling device was made on the the basis of the characters ofGaussian beam and its transformation characters through an aspheric lens, which canachieve the coupling efficiency of50%for a100m single-mode fiber with a core diameter of3.5m and a numerical aperture of0.13.
2. An underwater optical probe, a watertight chamber housing the optical sensingsystem, was developed. The laser transmitted by a single-mode fiber entered thechamber, then split into two parallel beams and intersected with each other outside thechamber. The beam intersection defines the measurement volume in the seawater. Thedirections of±90o deviating from the transmission direction of laser beams arerespectively chosen to receive laser-induced fluorescence and laser Doppler signals inorder to keep both signals synchronic to get accurate size distribution. A robustbeam-splitting device, with output intensity insensitive to the incident beam directionchange caused by slight incident inclination and sea waves, was designed, using alateral displacement beamsplitter. Lab and field experiments proved that the opticalprobe chamber with its support system met the measurement requriements.
3. Photoelectric detection circuits and data acquisition&processing hardwareswere designed. DSP programs&PC software units for data processing, storage anddisplay were designed and completed.
4．The measurement system prototype was debugged, tested, further adjusted inthe laboratory, and the error analysis was done. In the laboratory tests and calibrationexperiments,12phytoplankton species in3different kinds of shapes cultured in the labwere used. Gaussian curve-fitting method was adopted to study the phytoplankton sizedistribution data. The results of lab experiments were compared with that measured byCASY-TT cell counter+analyser system. Corresponding size distribution andconcentration errors for all kinds of phytoplankton measured in the tests were given.Statistical distribution errors of size distributions for spherical or ellipticalphytoplankton with a size range of5-50m were less than10%. The average size valuemeasured by the prototye was well linearly relevant with the one measured byCASY-TT, The concentration measurement errors of mixed phytoplankton were around5%.
5. The prototype in situ experiments were done twice at different stations inQingdao coastal waters, and the results proved that the designed system can be used for in situ measurement of size distribution and concentration information of marinephytoplankton, The adjusted system can also do real-time monitoring at a fixedposition for a long time. The experimental data demonstrated no distinct change ofphytoplankton concentration in the measured waters. Compared with microscopesobservation results, the measurement errors are within an acceptable range.
The system developed in the thesis is capable of real-time size distribution andconcentration measurement for phytoplankton particles. The underwater opticalsensing probe does not rely on electrical power, instead, it uses optical fibers totransmit laser and the measured optical signals. The sensing probe can stay submergedfor long period of time, making a long-term real-time in situ monitoring possible. In thesame time, compared with the other measurement methods, the system needs neithercomplicated optics nor complex algorithms, so it is of outstanding real-timeperformance. The system has a promising application in marine environmentmonitoring and evaluation,“red tide” research, marine ecosystem process research andwater quality monitoring.
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