Environmental Sample Processor

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

9. 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

11. 2G ESP • Weighs over 800lbs • Positively buoyant • Radio modem on a surface float

12. A railroad wheel is used as an anchor

13. Contains pucks in which samples are analyzed

14. 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

15. 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

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

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

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

20. 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.

21. 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.

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

24. 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