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Optical sensing of plankton communities and dynamics
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FluoSieve: A novel fluorescence imaging flow cytometer for high-throughput phytoplankton analysis
Tuesday 8th @ 1030-1050, Conference Room 7
Jianping Li* , Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences
Mark Luk, Xcube Technology Ltd
Tao Chen, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences
Peng Liu, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences
Qinmu Peng, Huazhong University of Science and Technology
Presenter Email: jp.li@siat.ac.cn |
| High-throughput quantitation and characterization of phytoplankton in natural seawater is of fundamental significance for oceanography sciences and environmental monitoring. Imaging flow cytometry (IFC), with FlowCAM and IFCB as representative instruments, can extract multi-parameter statistical information of water samples by analyzing numerous phytoplankton images captured while they flow through an optical interrogation area, thanks to its high-throughput capability in acquiring phytoplankton images with cellular resolution and processing digital images for quantification and classification. However, taking fast yet accurate measurement of diverse natural phytoplankton with extreme heterogeneity remains challenging for current IFC instruments. Some fundamental issues such as lack of sensitivity and resolution for detecting picophytoplankton, compromises between imaging throughput and imaging quality due to motion/defocusing blurring for analyzing larger microphytoplankton, and trade-off between analyzing throughput in terms of water sample volume with statistical accuracy, still limited their application in practice especially in field scenarios, such as on a research vessel or in a coastal observatory, where autonomous and long-time operation is preferred.Combining light-sheet microscopy with axial flow-through detection, a new fluorescence IFC named FluoSieve has been devised and developed recently. This new imaging sensor uses lasers as light source and has demonstrated several advantages over shortcomings associated with traditional IFCs for phytoplankton analysis. FluoSieve uses single objective lens with high magnification, suppresses much out-of-focus photon noise, has higher excitation power density and integrates more in-focus photons, all together facilitate great enhancement in phytoplankton imaging quality in terms of resolution, signal-to-noise, size coverage and throughput. Based on foundations built in laboratory settings, we have upgraded FluoSieve from an indoors bench-top prototype into a field applicable instrument system recently. This new development is to promote the technology towards more field applications, such as time series observation on coastal stations and underway observation onboard a research vessel. In this talk, we will report the instrumentation progress and preliminary measurements. Through these developments and application collaboration with oceanography scientists, the FluoSieve technology is anticipated to find its niche in more applications and that demand automated phytoplankton analysis. |
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