1. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Fluorescent probes of life: Delivery system Life in the Atacama Design Review December 19, 2003 Waggoner CMU

2. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Description and Motivation Fluorescent probe delivery system Must place 4 different dyes within cell structures of biofilm or spores or exposed cells

3. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Key Requirements Place 4 fluorescent probes on DNA, protein, lipid and carbohydrate components of cells - Softening of cell encapsulation structures - Delivery of probes - Possible wash step Delivery to exposed rock or soil below camera Good coverage without getting probes in detection optics Robust, low energy, small size

4. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Design Considerations Three containers for spritzed components Nozzles for directed spray Corrosion-resistant strong tubing Pressure or pump delivery system Pump requires energy – est. 10W for 3 min Alternative compressed air with switches Sufficient dye must be delivered to cell components – timing and flow rate – Rover electronics Carrier solvents for the three sprays The fluorescent probe selection – separate design

5. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Technical approach Spritz 1: Sample preparation - Soften secreted coatings, permeabilize cells - Incubate 5 min to 2 hr – to be determined Spritz 2: Fluorescent probe delivery - Probably water-ethylene glycol mix - Incubate 5 min to 1 hr – to be determined Spritz 3: Wash excess probe away

6. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Implementation Issues Are prototypes to be tested? Manual method works What are the potential failure conditions? Clogging Wind blows spray away or into optics Electronic system fails What are the major schedule issues? Need person for this work – 2d/wk, 4 wks Manual testing with chemistry/biology – 20d System needs to be designed for rover – electronics, mechanical, space on rover

7. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Cartoon of spritz system Pumps Conditioning Wash 4 Fluor probes Nozzles Pump controller Rover control Rock

8. Life in the Atacama, Design Review, December 19, 2003 Carnegie Mellon Fluorescent probes DyeAbsorption max (nm)Emission max (nm)Enhancement on binding DNA probes SYTO 9470530++++ SYTO BC480500++++ DRAQ56506700 Protein probes Fluorescamine380464++++ OPA/2ME334455+++ NDA/CN419493+++ CBQCA/CN465560+++ SYPRO Orange470570 +++ SYPRO Red550630++++ Lipid probes DPH353449+++ Nile Red559637+++ DiI(7)750780+ Carbohydrate probe Calcofluor White 360480+++