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Biological and Chemical Sensor Workshop July 13-16, 2003 Fluorescence Spectroscopy in Oceanography Paula G. Coble College of Marine Science University of South Florida
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Biological and Chemical Sensor Workshop July 13-16, 2003 Acknowledgements ONR Grant #N00014-96-1-5010 ONR Grant #N00014-96-1-5010 Visanthi Sivaprakasan Visanthi Sivaprakasan Dennis Killinger Dennis Killinger Jennifer Boehme Jennifer Boehme Robyn Conmy Robyn Conmy Colin Stedmon Colin Stedmon Bob Chen Bob Chen
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Biological and Chemical Sensor Workshop July 13-16, 2003 Outline Definitions Definitions Applications Applications Systems and system components Systems and system components Data processing Data processing What’s next? What’s next?
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Biological and Chemical Sensor Workshop July 13-16, 2003 What is fluorescence? Emission of light from an excited state of a molecule Native fluorescence – no dyes added
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Biological and Chemical Sensor Workshop July 13-16, 2003 Position of emission maximum Position of excitation maximum Fluorescence intensity Peak width Steady State Fluorescence Parameters
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Biological and Chemical Sensor Workshop July 13-16, 2003 Chemicals to be sensed porphyrins porphyrins chlorophylls chlorophylls phycobilins phycobilins proteins proteins tryptophan tryptophan tyrosine tyrosine vitamins and coenzymes vitamins and coenzymes NADH NADH flavins flavins pterins pterins Chemical specificity, but not source specificity – produced by all living organisms
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Biological and Chemical Sensor Workshop July 13-16, 2003 Chemicals to be sensed porphyrins porphyrins proteins proteins vitamins, coenzymes vitamins, coenzymes CDOM, humic substance CDOM, humic substance PAHs PAHs plastics plastics Quinine sulfate Quinine sulfate
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Biological and Chemical Sensor Workshop July 13-16, 2003 Chemicals to be sensed porphyrins porphyrins proteins proteins vitamins vitamins CDOM CDOM PAHs PAHs plastics plastics phthalates phthalates bis-phenol A bis-phenol A SeawaterOil Contamination Epoxy CoatingChrysene
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Biological and Chemical Sensor Workshop July 13-16, 2003 Applications Spring Water Drinking water Drinking water organics organics contaminants contaminants bacteria bacteria bis-phenol A bis-phenol A
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Biological and Chemical Sensor Workshop July 13-16, 2003 Applications Drinking water Drinking water Groundwater & soils Groundwater & soils PAHs PAHs organic contaminants organic contaminants petroleum petroleum Al-Awadi et al., 2001 Kuwait oil fields
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Biological and Chemical Sensor Workshop July 13-16, 2003 Applications Drinking water Drinking water Groundwater & soils Groundwater & soils Tumor detection Tumor detection NADH NADH Elastin, collagen Elastin, collagen tryptophan, tyrosine tryptophan, tyrosine Pradhan et al. 1995 NADH trp
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Biological and Chemical Sensor Workshop July 13-16, 2003 Applications Drinking water Drinking water Groundwater & soils Groundwater & soils Tumor detection Tumor detection Sediments Sediments chlorins chlorins PAHs PAHs Chen, Jiang & Zhao, 2000 chlorins
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Biological and Chemical Sensor Workshop July 13-16, 2003 Applications Drinking water Drinking water Groundwater (Kuwait oil fields) & soils Groundwater (Kuwait oil fields) & soils Tumor detection Tumor detection Sediments Sediments Forensics Forensics Ballast water Ballast water Oil spills Oil spills
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Biological and Chemical Sensor Workshop July 13-16, 2003 Techniques Steady state Steady state Time-resolved (fluorescence lifetimes) Time-resolved (fluorescence lifetimes) Front face: solid phase or solution Front face: solid phase or solution Remote sensing Remote sensing Rudnick and Chen 1998 SS TR pyrene
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Biological and Chemical Sensor Workshop July 13-16, 2003 Techniques Steady state Steady state Time-resolved (fluorescence lifetimes) Time-resolved (fluorescence lifetimes) Front face: solid phase or solution Front face: solid phase or solution Remote sensing Remote sensing Rudnick and Chen 1998
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Biological and Chemical Sensor Workshop July 13-16, 2003 Techniques Steady state Steady state Time-resolved (fluorescence lifetimes) Time-resolved (fluorescence lifetimes) Front face: solid phase or solution Front face: solid phase or solution Remote sensing Remote sensing Active Lidar Active Lidar Passive solar Passive solar Hoge et al.
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Biological and Chemical Sensor Workshop July 13-16, 2003 Steady state Steady state Time-resolved (fluorescence lifetimes) Time-resolved (fluorescence lifetimes) Front face: solid phase or solution Front face: solid phase or solution Remote sensing Remote sensing Multi-spectral, fluorescence contouring Multi-spectral, fluorescence contouring Techniques
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Biological and Chemical Sensor Workshop July 13-16, 2003 Fresh Riverine Very bleached Partially bleached
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Biological and Chemical Sensor Workshop July 13-16, 2003 Dual source dual detector spectrofluorometer Disadvantages Large size Large size Bench top Bench top Analysis time Analysis time Sensitivity Sensitivity Data processing Data processing
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Biological and Chemical Sensor Workshop July 13-16, 2003 -Measures six excitation & sixteen emission wavelengths -Operates in continuous underway or profiling mode In Situ Measurement of CDOM WetLabs SAFIre
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Biological and Chemical Sensor Workshop July 13-16, 2003 SAFIre OPTICS LAYOUT FLASH LAMP APERTURE INTERFERENCE FILTER PLANO-CONVEX LENS BEAM SPLITTER/ REFERENCE DETECTOR PRESSURE WINDOW FLOW TUBE ABSORPTION DETECTOR PRESSURE WINDOW/ DIFFUSER DETECTOR ARRAY
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Biological and Chemical Sensor Workshop July 13-16, 2003 CDOM fluorescence ratio 375/400 nm @ Ex = 307 nm Aug. 1998 Dec. 1998 Jan. 1999 high = bleached low = rivers
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Biological and Chemical Sensor Workshop July 13-16, 2003 Laser Sensor Development for Fluorescence Detection of Plastics and other Anthropogenic Compounds Dissolved in Seawater increase in sensitivity by factor of 80 to 1000 over conventional spectrofluorometer instruments; doubled number of optical channels. increase in sensitivity by factor of 80 to 1000 over conventional spectrofluorometer instruments; doubled number of optical channels. Portable LIF system Fluorescence signature during Ocean field test into Gulf of Mexico
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Biological and Chemical Sensor Workshop July 13-16, 2003
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Principal Component Analysis (PCA) Principal Component Analysis (PCA) Parallel Factor Analysis (PARAFAC) Parallel Factor Analysis (PARAFAC) Multivariate Analysis
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Biological and Chemical Sensor Workshop July 13-16, 2003 PCA Gulf of Mexico Endmembers Oligotrophic Atchafalaya Mississippi EcoHAB Coastal EcoHAB Offshore PC1 PC2 Boehme and Coble 2003
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Biological and Chemical Sensor Workshop July 13-16, 2003 PARAFAC Model (Stedmon, Markager, and Bro; 2003) x = fluorescence intensity x = fluorescence intensity f = analyte f = analyte i = sample i = sample j = emission j = emission k = excitation k = excitation a concentration a concentration b fluorescence quantum efficiency b fluorescence quantum efficiency c molar absorbtivity c molar absorbtivity
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Biological and Chemical Sensor Workshop July 13-16, 2003 PARAFAC Example 96.6 – 99.9% variability explained 96.6 – 99.9% variability explained Stedmon, Markager, & Bro 2003
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Biological and Chemical Sensor Workshop July 13-16, 2003 PARAFAC Components in Danish Waters 1-4 Terrestrial origin 1-4 Terrestrial origin 5 autocthonous 5 autocthonous Stedmon, Markager, & Bro 2003
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Biological and Chemical Sensor Workshop July 13-16, 2003 Fluorescence Imaging Mazel et al., 2003 Laser Line Scanner (PHILLS) Laser Line Scanner (PHILLS) Sediment infauna sections Sediment infauna sections Plankton community? Plankton community? False color coral head
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Biological and Chemical Sensor Workshop July 13-16, 2003 Microchannel Plate Detectors – multispectral PMT Pankrantz 1998
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Biological and Chemical Sensor Workshop July 13-16, 2003 Summary Cost and size of laser instruments is still decreasing Cost and size of laser instruments is still decreasing Multispectral sensors increasingly sensitive and selective Multispectral sensors increasingly sensitive and selective Fiber optics expand flexibility to harsh/inaccessible environments Fiber optics expand flexibility to harsh/inaccessible environments Biomedical field will continue to drive instrument development Biomedical field will continue to drive instrument development Signal processing into discrete components (PARAFAC) Signal processing into discrete components (PARAFAC)
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