1. November 29, 2005 1 Frankie Jernigan MSG Integration Investigation Operations Lead EO30 Frankie.r.jernigan@nasa.gov Frankie.r.jernigan@nasa.gov 256-544-2487 Protein Crystal Growth Monitoring by Digital Holographic Microscope (PromISS) PromISS 2 Hardware Processing PromISS 4 Samples

2. November 29, 2005 2 Class Objectives Crew member will be able to assemble experiment. Crew member will know what photographs to acquire. Crew member will understand experiment operational flow, hazards and mitigation.

3. November 29, 2005 3 Experiment Objectives To characterize the time crystallization experiments will take to reach equilibration in a microgravity environment using the counter diffusion crystallization method. To produce protein crystals for ground-based X-ray diffraction studies. To use the holographic microscope in the PromISS 2 Experiment Box to capture (via video) and measure several parameters of the growing protein crystals.

4. November 29, 2005 4 Science Overview The monitoring of protein crystal growth in a three-dimensional environment to produce a detailed analysis and quantitative interpretation of the relationship between the quality of the crystals and the environment in which they were produced. Strategic Objective: understanding the basic processes underlying the behavior of proteins at high concentrations. Strategic Objective Mapping: Since the whole process of crystal growth from solution is controlled by the strength of the driving force (super saturation), a tractable scenario for the study of mass transport and its effect on crystal growth and crystal quality would contribute to many fields such as macromolecular crystallography, materials processing, etc. Space Applications: Development of microgravity-tailored techniques for producing crystals of better quality or size, eventually allowing polymorphic selection or the production of new polymorphs. Earth Applications: A basic comprehension of the processes leading to crystals having the desired properties is required to asses the feasibility of any given innovation in on-ground crystallization techniques. Once the relative importance of mass transport or growth rate steadiness, crystal and fluid movements, etc. is known, new on-ground techniques can be devised to get the best possible on-ground crystals and to asses the requirements and expected benefits of microgravity experiments in the field.

5. November 29, 2005 5 PromISS 4 OPERATIONAL FLOW PromISS SETUP Part 2 (10 min) Steps 4 to 4.34 Part 3 (35 min) Steps 4.35 to end. 45 Min Install PromISS 2 hardware PromISS 2 EXPERIMENT BOX PromISS 2 ELECTRONICS BOX Mate POWER and DATA cable assemblies Install MSG Camera onto PromISS 2 EXPERIMENT BOX ASSUMPTIONS 1.Hardware transfer completed, temporarily stowed 2.MSG Activated (MAIN PWR ON) 3.WV ILLUM – OFF 4.MSG VIDEO System – ON, (includes minimum power for; 1 CAMERA, 1 RECORDER, 2 MONITORS) Assumptions:1, 2 PromISS SETUP Part 1 (35 min) Steps 1 to 3 35 Min PromISS SAMPLE SAMPLE CONDITIONING 30 Min Install PromISS 4 Return Container 1 in MSG WV (12 hours prior to Ops) Install PromISS 4 Return Container 2 in Kubik Install PromISS 4 Sample Wheel, Sample Cells, and tape 1. Take pictures of entire sample wheel and each sample cell installed on sample wheel prior to installation in EXPERIMENT BOX. Assumptions: 1, 2, 3 Before 20 Progress Dock At least 12 hours required for Sample Conditioning. PromISS Software Upload 15 Min PromISS Software Upload 15 Min 1 hour and 30 min command window between.

6. November 29, 2005 6 PromISS ACTIVATION 10 Min AIR CIRC – ON (Default MODE 4) Power On PWR OUT 2 +5V,  12V, +28V MSG VIDEO System* PromISS 4 PROCESSING Unattended 15 Days Once PromISS is activated it will go to a “warm up” and run continuously The payload software will send commands to the MSG Video System PromISS includes one MSG Camera mounted internally. Camera power is on for the entire processing time of 15 days. AIR CIRC will be ON ASSUMPTIONS 1.Hardware transfer completed, temporarily stowed 2.MSG Activated (MAIN PWR ON) 3.WV ILLUM – OFF 4.MSG VIDEO System – ON, (includes minimum power for; 1 CAMERA, 1 RECORDER, 2 MONITORS) Assumptions: 2, 3, 4 Assumptions: 2, 3, 4, PromISS VIDEO OPERATIONS 10 Min Repeated 7 times Exchange PromISS 4 tapes 1 through 7, install 8 PromISS 4 OPERATIONAL FLOW (cont.) PromISS Software Upload 30 Min AIR CIRC – ON (Default MODE 4) MSG VIDEO System* Assumptions: 2, 3, 4

7. November 29, 2005 7 PromISS SAMPLE EXCHANGE Part 1 (20 min) steps 1-2 Part 2 (30 min) steps 3-4 50 Min Assumptions: 1, 2, 3 Remove tape 8, stop processing, power down, remove PromISS 4 Sample Wheel, take pictures of the entire wheel and each cell, stow in PromISS 4 Return Container 1, and stow Container 1 in Kubik. PromISS 4 OPERATIONAL FLOW (cont.) Assumptions: 1, 2 PromISS SAMPLE CONDITIONING 2 30 Min PromISS SAMPLE EXCHANGE Part 3 (30 min) steps 5.-5.31 Part 4 (10 min) steps 5.32-end Assumptions: 1, 2, 3 Install second set of PromISS 4 Sample Cells on Sample Wheel, take picture of entire wheel and of each cell, install tape 9 and install in EXPERIMENT BOX Install PromISS 4 Return Container 2 in MSG WV (6 hours prior to Ops) 40 Min ASSUMPTIONS 1.Hardware transfer completed, temporarily stowed 2.MSG Activated (MAIN PWR ON) 3.WV ILLUM – OFF 4.MSG VIDEO System – ON, (includes minimum power for; 1 CAMERA, 1 RECORDER, 2 MONITORS) At least 6 hours required for Sample Conditioning.

8. November 29, 2005 8 PromISS STOW Part 1 (20 min.) steps 1 – 2 Part 2 (15 min.) Steps 3 - 4 Part 3 (20 min.) steps 5-7 Power Off PWR OUT 2 +5V,  12V, +28V VIDEO System – STBY(remove TAPE 16) Remove Sample Wheel, take pictures of each Sample Cell, remove SAMPLE CELLS place into RETURN CONTAINER 2 AIR CIRC – OFF Remove, temporarily stow PromISS 2 hardware MSG MAIN PWR – STBY Stow Return Container 2 in Kubik 55 Min PromISS 4 PROCESSING Unattended 15 Days Once PromISS is activated it will go to a “warm up” and run continuously The payload software will send commands to the MSG Video System PromISS includes one MSG Camera mounted internally. Camera power is on for the entire processing time of 15 days. AIR CIRC will be ON Assumptions: 2, 3, 4, PromISS VIDEO OPERATIONS 10 Min PromISS ACTIVATION 10 Min AIR CIRC – ON (Default MODE 4) Power On PWR OUT 2 +5V,  12V, +28V MSG VIDEO System* Assumptions: 2, 3, 4 Repeated 7 times Exchange PromISS 4 tapes 9 through 15, install 16 PromISS 4 OPERATIONAL FLOW (cont.) ASSUMPTIONS 1.Hardware transfer completed, temporarily stowed 2.MSG Activated (MAIN PWR ON) 3.WV ILLUM – OFF 4.MSG VIDEO System – ON, (includes minimum power for; 1 CAMERA, 1 RECORDER, 2 MONITORS)

9. November 29, 2005 9 PromISS setup inside MSG Work Volume PromISS 2 Locking System PromISS 4 Return Container PromISS 2 Electronics Box PromISS 2 Experiment Box Back Operations Flow

10. November 29, 2005 10 PromISS 2 Hardware on board ISS PromISS 2 Kit 1 PromISS 2 Electronics Box with pigtailed harness (see slide number 12) PromISS 2 Kit 2 PromISS 2 Clamp (see slide number 11) PromISS 2 Sample Wheel (not used for PromISS 4 Operations) PromISS 2 Software Upload Cable (see slide number 12) PromISS 2 Locking System (see slide number 15) PromISS 2 Experiment Box This hardware is assembled in MSG prior to 20 Progress Dock in the following order. PromISS 2 Electronics Box PromISS 2 Experiment Box Cable Connections MSG Video Hardware

11. November 29, 2005 11 PromISS 2 Kit 1 PromISS 2 Kit 2 PromISS 2 Kit 1 contents: PromISS 2 Electronics Box with pigtailed harness PromISS 2 Kit 2 contents: PromISS 2 Clamp PromISS 2 Sample Wheel PromISS 2 Software Upload Cable PromISS 2 Locking System

12. November 29, 2005 12 PromISS 2 ELECTRONICS BOX PromISS 2 SOFTWARE UPLOAD cable PromISS 2 ELECTRONICS BOX External Components (accessible by crew) - One momentary switch - Pigtailed harness PromISS 2 SOFTWARE UPLOAD cable Is used to verify the software integrity and upload new software to the Electronics Box.

13. November 29, 2005 13 PromISS 2 EXPERIMENT BOX External Components (accessible by crew) - Two momentary switches -Three LEDs - Camera mount - One POWER/DATA connector - Cover Lid

14. November 29, 2005 14 PromISS 2 Camera Mount External Components (accessible by crew) –√ Kapton Tape removed from the viewing window on the top left rear corner of the PromISS 2 Experiment Box. –The MSG Camera attaches to the camera mount on the top left rear corner of the PromISS 2 Experiment Box.

15. November 29, 2005 15 PromISS 2 Locking System Part A Part B

16. November 29, 2005 16 PromISS 2 Hardware launched on 20 Progress PromISS 4 Kit 1 PromISS 4 Tape 1-8 (see slide number 23) PromISS 4 Return Container 1(see slide number 18) PromISS 4 Sample Cell 0-5 (see slide number 21) PromISS 4 Activation Tool (see slide number 20) PromISS 4 Balltip L-Wrench (see slide number 20) PromISS 4 Sample Wheel (see slide number 21) PromISS 4 Kit 2 PromISS 4 Tape 9-16 (see slide number 23) PromISS 4 Return Container 2 (see slide number 18) PromISS 4 Sample Cell 6-11 (see slide number 21) Kubik (utilized during PromISS operations) (see slide number 24) This hardware is assembled in MSG after 20 Progress Dock in the following order. PromISS 4 Return Container 1PromISS 4 Return Container 2 PromISS 4 Activation ToolPromISS 4 Sample Cell 6-11 PromISS 4 Balltip L-WrenchPromISS 4 Tape 9-16 PromISS 4 Sample Wheel PromISS 4 Sample Cell 0-5 PromISS 4 Tape 1-8

17. November 29, 2005 17 PromISS 4 Kit 1 PromISS 4 Kit 2 PromISS 4 Kit 1 contents: PromISS 4 Tape 1-8 PromISS 4 Return Container 1 PromISS 4 Sample Cell 0-5 PromISS 4 Activation Tool PromISS 4 Balltip L-Wrench PromISS 4 Sample Wheel PromISS 4 Kit 2 contents: PromISS 4 Tape 9-16 PromISS 4 Return Container 2 PromISS 4 Sample Cell 6-11

18. November 29, 2005 18 PromISS 4 Return Container Components (accessible by crew) - Pressure Valve - Ten fasteners

19. November 29, 2005 19 PromISS 4 Return Container Cont. PromISS 4 Return Container is stowed in a zip lock bag in a kit. It is installed on the MSG coldplate during sample conditioning and sample installations. It is installed on the MSG rear wall during operations.

20. November 29, 2005 20 PromISS 4 Activation Tool PromISS 4 Balltip L-Wrench PromISS 4 Balltip L-Wrench is used to open the Return Containers and Experiment Box Lid. PromISS 4 Activation Tool is used to activate the PromISS 4 Sample Cells. It is combined with the ISS ¼” Driver Handle, 10mm Deep Socket.

21. November 29, 2005 21 PromISS 4 SAMPLE CELL PromISS 4 SAMPLE WHEEL Back Operations Flow

22. November 29, 2005 22 PromISS 4 SAMPLE CELL The above picture is of the PromISS 2 Sample Wheel holding PromISS 3 Sample Cells during PromISS 3 Operations. PromISS 2 Locking System installed PromISS 4 Sample Wheel is installed on the rear wall of the MSG using the PromISS 2 Locking System. The PromISS 4 Sample Cells are activated using the PromISS 4 Activation Tool and installed on the PromISS 4 Sample Wheel. Closed: The plunger is pressing against the middle part of the housing. Open: The plunger is fully retracted against the outside of the housing. Turning the plunger screw RIGHT (CW) will OPEN the valve.

23. November 29, 2005 23 PromISS 4 TAPES PromISS uses 16 digital tapes during the 30 days of operations. Eight tapes per 15 day run. Tapes are packed eight per kit.

24. November 29, 2005 24 Kubik Kubik is an incubator that will be flown on 20P and installed in the Russian segment. It will be used to keep the PromISS 4 samples at 22°C prior to and after processing in MSG.

25. November 29, 2005 25 PromISS 2 Experiment Box The two momentary switches WAIT TAPE will halt the sample wheel rotation and stop recording for tape exchanges, operator will toggle once to the up position prior to tape removal (LED illuminates green), and then up after tape exchange is complete (LED extinguishes, RUNNING LED illuminates) END RUN will initiate the PromISS 4 SAMPLE WHEEL rotation to the zero position. When toggled to the up position, the END RUN LED will illuminate and remain green until the PromISS 4 SAMPLE WHEEL has reached the zero position. Depending on the position of the PromISS 4 SAMPLE WHEEL when the switch is toggled, the time to zero position may take up to 1 hour. POWER/DATA Connector POWER/DATA (J308) will provide the power and data interface between MSG Facility and the PromISS 2 EXPERIMENT BOX PromISS Operational Notes

26. November 29, 2005 26 Cover Lid Provides for one level of containment for the PromISS 4 Sample Cells when properly sealed and contains two micro switches. The Experiment Box Lid must be properly closed in order for the micro switches to activate. Camera Mount The camera must be firmly seated within the mount or the micro switches that are in the bottom of the mount will not activate. During activation there is a check with the POIC for data confirmation that the micro switches are enabled. PromISS Operational Notes

27. November 29, 2005 27 PromISS 2 ELECTRONICS BOX The PromISS 2 ELECTRONICS BOX is fitted with three M6 Captive Screws that are identical to the ones on the PROMISS 2 EXPERIMENT BOX. There are three pigtailed harnesses protruding from the PromISS 2 ELECTRONICS BOX: POWER CABLE  MSG (P325) DATA CABLE  MSG (P311) POWER AND DATA CABLE  PROMISS 2 EXPERIMENT BOX (P308). The PromISS 2 ELECTRONICS BOX is also fitted with one permanent toggle switch (either OPERATION or UPLOAD mode) that enables the Crew to perform a software upload, or to operate normally. The toggle switch is protected by switch guards. PromISS Operational Notes

28. November 29, 2005 28 Two levels of containment are maintained during all PromISS operations. PromISS 4 Return Container 1 and 2 provide a level of containment. PromISS 4 Sample Cells provide a level of containment. PromISS 2 Experiment Box provides a level of containment MSG Air Circ provides a level of containment. If a leak is detected then the procedures provide the cleanup steps or direct the crew to the PromISS Cleanup Procedure. All necessary cleaning equipment is in the MSG Work Volume. In the event of a water cooling loop failure, the PromISS 2 ELECTRONICS BOX may exceed touch temperature limit values. If water cooling loop failure is detected the MSG Facility will automatically power down the payloads and the operator must wait a minimum of 10 minutes before entering the WV. The PromISS 2 ELECTRONICS BOX provides a High Voltage source to the PromISS 2 EXPERIMENT BOX via connector J308. Connector J308 is inaccessible at all times after PromISS Activation, precluding a potential shock hazard to the operator. Safety Notes PromISS 4 Samples are a TOX Level 1