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Environmental Sample Processor

Environmental Sample Processor. Developed by Monterey Bay Aquarium Research Institute (MBARI) Developmental team led by Chris Scholin. Purpose. Molecular analysis of samples taken in the ocean. 1G ESP. First generation ESP Developed in 1999 Used until 2006. First deployment. May 2001

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Environmental Sample Processor

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  1. Environmental Sample Processor

  2. Developed by Monterey Bay Aquarium Research Institute (MBARI) • Developmental team led by Chris Scholin

  3. Purpose • Molecular analysis of samples taken in the ocean

  4. 1G ESP • First generation ESP • Developed in 1999 • Used until 2006

  5. First deployment • May 2001 • Gulf of Maine • Recorded data on depth, temperature, salinity, chlorophyll fluorescence • Tested prototype DNA probe arrays

  6. Searching for Alexandruim fundyense- toxic dinoflagellate

  7. 2nd deployment • October 2002 • Monterey Bay • Different DNA probes used to look at harmful algae blooms and RNA from barnacles and diatoms

  8. 2G ESP • 2nd generation ESP started development in 2003 • Deployed Spring 2006 in Monterey Bay • Easier to service, lower power requirements, compact, and user-friendly

  9. 2G ESP • Weighs over 800lbs • Positively buoyant • Radio modem on a surface float

  10. A railroad wheel is used as an anchor

  11. Contains pucks in which samples are analyzed

  12. Pucks • Pop-top – for imaging a DNA array • Wet-storage – built in valve allows fluid to be stored • Dry-storage – to store samples which may be dried • Filter – to clean lysis prior to moving fluid to processing

  13. Pucks • Held by a circular array of 7 tubes called the carousel • When the ESP is deployed, 6 of the tubes are filled with pucks, as pucks are used they are dropped into the empty tube

  14. Major components of the 2G ESP • Core ESP • External Sampling modules • Analytical modules

  15. Core ESP • Housing • Power • Reagents • Data/control communications • Collects samples from milliliters to 2 liters • Site of automated sample processing

  16. Core ESP • 2 real time DNA probe arrays • Sandwich hybridization assay (SHA) • Enzyme-linked immunosorbent assay (ELISA)

  17. Core ESP • Sandwich hybridization assay • Collects a sample of water and creates a nucleic acid extract • Extract pumped into a puck and washed over the probe array • Another solution washed across the array with another probe • This completes the sandwich probe, target, probe • The probe generates a signal in the form of light when the target is present.

  18. Core ESP • Enzyme-Linked immunosorbent assay (ELISA) • Detects antigens and antibodies in a sample • Collects a sample of water and creates an extract mixed with an antibody it is searching for. • The extract it delivered onto the array surface. • If no toxin, antibody gets bound to array, if toxin is present, antibody gets bound to the toxin and is washed away. • Then an enzyme solution is used to create a signal to determine if the toxin is present.

  19. External Sampling Modules • Deep water sampling module • Allows for sampling deep water environments • Being developed to be deployed on the MARS cabled observatory

  20. Analytical Modules • Microfluidic block • Fluid handling station that distributes samples and reagents to one or more detection modules • PCR • Amplifies DNA • Walt Array • Uses epiflourescence to count bacteria in water samples

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