1. HAB Case Studies Current Automated Capabilities & Future Needs Automated submersible flow cytometry Flexible targets, ecological context (MVCO examples) Counterpoint to highly focused, specific molecular approaches HAB Examples in Monterey Bay, Gulf of Mexico, and Gulf or Maine A few thoughts on complementary and evolving combinations with other observing technologies > Observing platforms strategic fixed location  adaptive, mobile > Sensor targets bulk water – single cells – molecules > Sampling statistics detection of molecules, detection of cells space / time extent and resolution

2. Automated features for extended deployment Standard analysis, biofouling control, realtime humidity sensing & intake valve control  Routine ~6 month unattended deployments Imaging FlowCytobot Principles from conventional flow cytometry but automated and submersible Combined with video imaging Optimized for large cells (~5-300  m) Olson and Sosik 2007, Sosik and Olson 2007 Observational capabilities combined with automated image classification Enumeration, identification, and cell sizing Thousands of individual phytoplankton

3. Phytoplankton Community Assessment at the Martha’s Vineyard Coastal Observatory Imaging FlowCytobot Microplankton

4. 27 diatom taxa at MVCO

5. Increasing overall rank 2007 2008 2009 Diatom diversity changes at MVCO – seasonal to interannual Top 20 diatom groups (pooled rank by biomass) 1 20

6. Monterey Bay – Dinoflagellate community analysis Akashiwo Akashiwo ( with dividing cells) Cochlodinium, etc. Unidentified dinos Prorocentrum & Dichtyocha 4-15 Sep 2006 Akashiwo 0 – 7400 cells mL -1

7. Karenia brevis UT Marine Sciences Inst pier Deployed since Sept 2007 Collaboration with Lisa Campbell

8. Karenia brevis Light scattering October 2009 Chl fluorescence Gulf of Mexico – Karenia bloom in 2009

9. Port Aransas area fish kills Oct. 2009 (Texas Parks and Wildlife Department; http://www.tpwd.state.tx.us) Gulf of Mexico – Karenia bloom in 2009 Daily resolution low level detection >1 month before peak Offshore origin 2-h resolution

10. Gulf of Mexico - Toxic (DSP) Dinophysis event in 2008 Abundance at 2-hr resolution microscopy (+ 95% CL) Unprecedented event, led to shellfish closures Campbell et al. 2010

11. Alexandrium Gulf of Maine – Alexandrium detection Brosnahan, Anderson et al. DNA fluorescence Species probe fluorescence Emerging imaging flow cytometry application Low biomass HAB  Automated image classification combined with species-specific nucleic acid probe Enumeration, plus life cycle studies

12. HAB Case Studies Current Automated Capabilities & Future Needs A few thoughts on complementary and evolving combinations with other observing technologies > Observing platforms strategic fixed location  adaptive, mobile > Sensor targets bulk water – single cells – molecules > Sampling statistics detection of molecules, detection of cells space / time extent and resolution

14. Detritus Chl Fluorescence Light Scatter 10 µm cells cryptophytes Flagellates Beads Dinophysis cf ovum Gulf of Mexico - Toxic (DSP) Dinophysis event in 2008

15. Karenia species – automated classification challenge