High Altitude Extremophiles

Transcription

1 High Altitude Extremophiles DESIGN DOCUMENT Team Phoenix Submitted by: Nicole Dudley Nathan Lester Robert Lossing Russell Morris Cory Smith Submitted to: Professor Koehler 18 April 2006 Phoenix - 1-5/4/2006

2 1.0 Mission Overview Design Management Budget Test Plan Experiment Expected Results Launch and Recovery Phoenix - 2-5/4/2006

3 1.0 Mission Overview Our primary mission is to create a balloon satellite which will contain an experiment to collect high altitude bacteria in hopes of determining which types of bacteria can survive in such extreme conditions. Astrobiology is the scientific study of the origin, distribution, evolution, and future of life in the universe. Our goal is to further the knowledge and understanding in this vital field by collecting samples of complex bacterium which are able to exist and even thrive in harsh environments such as high UV radiation, low temperatures and pressures. Bacteria found at such harsh environments would be very similar to possible bacteria found on other moon and planets like Mars, Europa, or Titan. Bacteria found on these planets would have to survive deadly UV radiation, freezing and boiling temperatures, and little or no pressure. Furthermore it is entirely possible for bacteria to live in space. In 1969 Apollo astronauts were surprised to discover microorganisms on the Surveyor probes which had landed on the moon three years earlier. The organisms had survived the launch, the harsh vacuum of space, three years of deadly exposure to radiation, being frozen at an average temperature of only 20 degrees above absolute zero, along with no nutrients, water or energy sources. Yet, they still survived. We want to collect and learn more about these fascinating organisms. There is still much to be learned of how they are able to survive in impossible environments. The possibilities gained by this knowledge are endless. We would be able to engineer an oxygen producing organism that would be able to survive the surface of other planets and oxygenate the surface. Knowledge gained from UV resistant bacteria can help benefit the treatments of cancers as well and make chemotherapy less dangerous. This is why we hope to collect and discover these hardy organisms for scientists to examine. 2.0 Design Heat The heat for the balloon satellite will be produced from a heating element powered by a 9V power source. The satellite will be insulated with foam that is approximately 1.5 cm thick. The joints will be sealed with aluminum tape to further reduce the loss of heat. 2.1 Design Camera The camera subsystem consists of two cameras, one facing the top on the balloon satellite and the other facing the bottom. Both of the cameras will be controlled using a Brainstem General Purpose 1.0 Module powered by a 12V power source. The signal will pass from the Brainstem to a relay which will trigger the cameras. The module will be programmed to take pictures more frequently as the mission time increases. Phoenix - 3-5/4/2006

4 2.3 Design Bacteria Collection The goal of the bacteria collection is to collect 3 separate samples at different times in the flight. To do this air is going to be drawn through glass tubes and will pass through a bacteria filter. 35 mm diameter fans will be used to draw the air through the filters. The fans will be controlled using the Brainstem Microcontroller. 15 minutes into the flight the first collector will be activated and remain activated for another 15 minutes. After which the first will turn off and the second will be activated for 15 minutes. The final experiment will also run for 15 minutes. The diagram below show how the experiment is going to look. Air will be drawn into the glass tubing from the top. The curve in the tubing is to stop bacteria from passing into the biofilter too early. The air will then pass through the bio-filter and be pushed out the bottom of the satellite by the fans. Phoenix - 4-5/4/2006

5 The next diagram shows how the Brainstem will be connected: The following diagram shows representations of the positions of the materials that are including the main experiment. These include the cameras, HOBO, brainstem, and power supplies. They will be placed inside our payload to reduce as much size as possible. The diagram is illustrated without the insulation and foam core. Phoenix - 5-5/4/2006

6 Our functional block diagram, shown below, describes our new and updated design. It depicts the idea of now using the brainstem to control both the cameras and the disk covering the Petri dishes. The heater requires its own power supply and the HOBO is internally powered. Also, because the HOBO is used with a time delay, a switch is not required to control it. Phoenix - 6-5/4/2006

7 Phoenix - 7-5/4/2006

8 3.0 Management Team Leader Robert Lossing Power and Heat Russell Morris Nathan Lester Optics Nicole Dudley Cory Smith Budget Nicole Dudley Experiment Robert Lossing Russell Morris Structures Cory Smith Nathan Lester Schedule: 2/21/2006 Order All Hardware 2/23/2006 Hardware Checkout 2/23/2006 In-Class Feedback 2/23/2006 6:30 PM Team Meeting (discuss design) 2/28/2006 In-Class Team Time 2/28/2006 Complete Satellite Design 2/28/2006 5:00 PM Team Meeting (work on Design Document Rev A) 3/2/2006 Design Document Rev A DUE 3/7/2006 In-Class Team Time 5:00 PM Team Meeting (discuss proposal and presentation 3/7/2006 feedback) 3/14/2006 In-Class Team Time 3/17/2006 5:00 Team Meeting (work on prototype) 3/16/2006 Prototype Built 3/20/2006 5:00 PM Team Meeting (work on Design Document Rev B) 3/21/2006 In-Class Team Time (work on CDR slides) 3/21/2006 CDR Complete 3/23/2006 Critical Design Review (presentations due at 12:00 PM) 3/23/2006 Design Document Rev B DUE 3/24/2006 Acquire All Hardware 4/4/2006 Programming and Circuitry Complete 4/6/2006 5:00 PM Team Meeting (assembly) Phoenix - 8-5/4/2006

9 4/6/2006 Integration of Camera 4/11/2006 Drop Test (with rocks of equivalent weight) 4/12/2006 Sterilize Glassware 4/13/2006 In-Class Feedback 4/13/2006 3:30 PM Team Meeting 4/13/2006 Satellite Complete 4/13/2006 Integration of Switches 4/13/2006 Integration of Hobo and Brainstem 4/13/2006 Integration of Heater 4/13/2006 Integration of Power 4/14/ :00 PM Team Meeting 4/14/2006 Whip Test 4/14/2006 Cold Test 1 (with cooler and dry ice) 4/15/2006 Cold Test 2 (with new batteries) 4/15/2006 Sterility Testing 1 4/16/2006 Sterility Testing 2 4/17/2006 Camera/Film Test 4/17/2006 5:00 PM Team Meeting (work on Design Document Rev C) 4/18/2006 Bring All Hardware to Class 4/18/2006 Design Document Rev C DUE 4/18/2006 Integration of Experiment 4/18/2006 Satellite Completely Finished/Ready to Fly 4/20/2006 Bring All Hardware to Class 4/20/2006 Pre-Launch Inspection 4/20/2006 Launch Readiness Review (presentations due at 12:00 PM) 4/20/2006 5:00 PM Team Meeting (finalize launch program) 4/21/2006 Final BalloonSat Weigh-in and TURN IN (by 1:30 PM) 4/22/2006 LAUNCH DAY (7:00 AM at Windsor, Co.) 4/23/2006 Data Collection Process 4/25/2006 Report and Data Analysis Guidance 4/27/2006 5:00 PM Team Meeting (data analysis) 4/27/2006 Compile Results 4/27/2006 Prepare for Final Presentation 4/29/2006 ITL Design Expo (12:00 PM - 4:00 PM) 5/1/2006 5:00 PM Team Meeting (final presentation practice/run through) 5/2/2006 Final Presentations (due at 12:00 PM) 5/3/2006 6:00 PM Team Meeting (work on Design Document Rev D) 5/6/2006 Design Document Rev D (final report) DUE Phoenix - 9-5/4/2006

10 4.0 Budget To help ensure that our budget is followed, each price and mass was placed at its highest possible value. Receipts will be kept and filed so that we can keep track of the exact money amount that we spend. Nicole will be in charge of the budget, so masses and money amounts will have to be approved by her before proceeding with the schedule. This will also help guarantee that both budgets will not be exceeded. Structures Cost Mass (g) Foam Core $ Insulation $ Adhesive $ Camera Camera with Film $ Film Developing $10.00 N/A Bacteria Experiment Brainstem $ Bacteria Filters (3) $ Glass Tubes (3 sets) $ Adapter $0.00 N/A Fans (3) $ Connectors and Crimps $28.98 Relays $12.11 Included in Brainstem mass Included in Brainstem mass Power and Heat 9V Batteries (6) $ V Batteries (3) $ Heater with Switch $ HOBO H $ Temperature Probe $ Testing Cooler $2.69 N/A Dry Ice $0.00 N/A Film/Developing $12.97 Total Cost Total Mass (g) $ Phoenix /4/2006

11 5.0 Test Plan We will be doing a number of tests to help ensure that our satellite will be ready for launch on April 22 nd. We plan to perform a camera and film test, drop test, whip test, experiment tests and finally, a cold test. For the camera and film test, we will first attach the camera to the brainstem and power supply. This will allow us to manipulate the timing intervals for each camera. Various small tests will have to be executed to make certain that the timing works correctly. The film part of this test will involve actually taking pictures from our satellite with a 12 exposure role and then developing the pictures. This will help gauge where the cameras must be placed on our balloon satellite and ensure that the cameras do in fact take successful pictures. The drop test will be performed by dropping the structure of our satellite from an overhang about 5 meters from the ground. We can then determine if it will stay intact upon impact during landing. During the actual flight, our satellite will be traveling approximately 14 m/s as it hits the ground. After performing this test we will observe the damage and discuss whether it is ready for launch. The whip tests purpose is to test the strength of the satellite s attachment to the string as it is tossed around during the decent. There will be massive forces that have the ability to tear the satellite apart from the string. To make sure it can withstand this, we will swing the satellite around and subject it to similar forces. It will be attached to a string and whipped and pulled in different directions and the potential damage will then be examined. In order to ensure that the collected bacteria is the bacteria gathered at the high altitudes and not bacteria contaminated from other sources, tests regarding the sterility of the science experiment subsystem must be completed. The experiment subsystem consists of the rotating disk that will expose the Petri dishes and the Petri dishes themselves. We plan to have two main tests and a control test. In order to test the sterility of the Bio-filter we will simulate the preflight setup to prove that bacteria are not contaminating the syringe filter prior to flight. This test will involve assembling the Bio-filter assembly and leaving it exposed to a simulated outdoor environment for 3 hours and then plating them on an agar growth Petri dish to observe the results. If the culture turns up negative for growth then the assembly design is sterile and contamination free. If the culture turns up positive for growth then the Bio-filter assembly needs to be reexamined. The second test is used once the design is considered sterile and the goal is to use the fan vacuum design to draw air and suspended bacteria through the filter once turned on at the desired altitude. This test will test the effectiveness of the vacuum created by the impeller fan design. The fan will be left running in a outdoor environment for 15 min to simulate the time allowed during flight for bacteria to be captured. Assuming that bacteria concentrations will be highest at ground level the growth results should be highest compared to bacteria concentrations at higher heights. The growth should show up on the agar Petri dish. The fan should draw in 2.1 cubic meters of air per 15 minutes. Using this data we can later determine the concentration of bacteria per cubic foot of air at the different altitude levels. The third test with the science experiment subsystem is simply a control test. For the control we will remove the rotating plate from the subsystem which will thus expose the Petri dishes to the air. During this test we hope to show how much growth would occur in the dishes if Phoenix /4/2006

12 the rotating dish were absent. This test will take approximately three hours and will hopefully show the progression of bacteria growth. When the satellite is fully functional, all the power systems will be switched on. The satellite will then be submerged in a Styrofoam cooler filled with dry ice for its final test. It will remain in there for the amount of time the real flight is likely to last and will be checked afterwards to make sure all systems are still up and running properly. This test will be our cold test. 6.0 Experiments The collection of high altitude bacteria in a scientific and professional way is a difficult task. Sterility techniques and decreasing the risk of unwanted contamination are two of the biggest concerns. Contamination is such a large threat that if even a single invisible bacteria cell contaminates the filters it will render the experiment useless. This is why we are following some of the strictest sterile technique methods to ensure a proper result. A method had to be devised to collect bacteria at only a certain time/altitude during the flight. Our earlier method of using a Parmesan rotating disk to uncover and cover Petri agar dishes turned out to be impossible to keep sterile. For our finalized experiment we ordered disposable gamma irradiated syringe filters. With the use of these sterile syringe filters and a simple vacuum set up, we will be able to take samples at exact times during our flight. Each one of the three Bio Filter Collectors will be fitted with a small 35 mm DC fan. By reversing the flow of the fan we are able to draw air through the filters like a vacuum. The 3 fans are controlled by a Brainstem Micro Controller. Upon launch the controller will activate the fans at specific times for 15 minute intervals during the flight to draw in air samples at the given altitudes. This collection technique is very light, simple, and easily sterilized. Once the balloon satellite returns to the earth the filters will be disconnected and sealed using hot sterile glue. They will then be placed in a cooler for transport to a lab for plating. To grow bacteria they first must be plated on agar Petri dishes. To get the bacteria off of the filters an irrigation syringe with sterile LB nutrient broth will be attached to the reverse side of the filter which will flush the bacteria off onto nutrient agar Petri dishes. Each filter will be flushed three times with 3cc of LB broth onto three separate Petri Dishes. The first plate should have the highest concentration of bacteria and the second with less and so on. The first plate with the most bacteria on it will be placed in the coldest freezer for growth. The second and third will be placed in successively warmer freezers. By doing this, we will be dividing the bacteria up into separate incubation temperatures in order to grow different bacteria that grow at different temperatures. This will allow us to separate out at which temperature each of the bacteria grows at. We hypothesize that the least amount of bacteria will grow in the coldest freezer. So we plan to use the plate with the highest concentration of bacteria on it. The bacteria will be grown in a cold environment to mimic their living conditions and only allow those bacteria to grow while inhibiting other bacteria. The bacteria will need to be on a specific oligotrophic (low food) media which is not as rich in nutrients as other bacteria as the goal is to grow oligotrophic bacteria. Once the bacteria have grown and multiplied we will be able to either look at them under a microscope or use a process called PCR (Polymerase Chain Reaction), which is a molecular biology technique for enzymatically copying DNA. The technique allows a small amount of DNA to be amplified many times, making more DNA available and analysis much easier. PCR is Phoenix /4/2006

13 commonly used in medical and biological labs for a variety of tasks, such as the detection of hereditary dieses, the identification of genetic finger prints, diagnosis of infectious diseases, cloning of genes, paternity testing, DNA computing, etc. Bacteria can also be observed using lab microscopes. We plan however to gain access to the University of Colorado s high powered electron microscope which will allow us to see a bacteria cell magnified 10,000 times, which is strong enough to see internal parts of a bacterium cell. 7.0 Expected Results After the completion of our April 22 nd flight we hope to have captured high altitude bacteria using our three Bio Filter Collectors. We look forward to growing colonies of these bacteria for further study and analysis. The bacteria we plan to discover will be extremophiles, or bacteria which can survive in harsh and deadly environments. By comparing the time at which the fans were activated and the ascent rate we will be able to approximately determine where each bacteria was collected from. Any collected bacteria that can be cultured will give us a glimpse into the adaptive abilities of complex life forms. The bacteria collected will have been able to survive the extremely low pressures and temperatures present at these altitudes. By finding bacteria at these altitudes we will be able to expand the limits at which life can survive. Also, the atmospheric conditions at these altitudes is fairly similar to the conditions found on the surface of our neighboring planet Mars. Using this knowledge, living bacteria found at these heights would help make the idea of life on Mars more plausible. The genes of recovered bacteria can be harnessed through research and possibly help produce powerful new medicines. Genes from UV resistant bacteria could provide UV resistance to other bacteria and even animals. Further studies for this project could be a global mapping of high altitude bacteria to determine global populations, migrations etc. Future projects should include stronger fans for a higher volume intake as well as a reusable filter design. There is really a tremendous amount of knowledge to be learned regarding this area of study. 8.0 Launch and Recovery Prior to launch all of the batteries and components will be connected and sealed into the cube. The morning of launch the switches will be flipped to activate the brainstem controller and heater. The preset program of the HOBO will activate on its own and continue until the flight is over. The protective aluminum foil covers will be removed from the sampling tubes prior to release. After recovery, our collected data will be analyzed. Our data will be in the form of bacteria which can be cultured in a laboratory. The bacteria will provide information in the form of unique DNA information that can be easily recovered, even from dead cells. Even if only a single bacteria cell is collected from our Petri dishes it can still be grown using rapid bacteria growth techniques in an incubator. Once the sample bacteria have been replicated they will be plated on a microscope slide. They will then be observed under a high powered microscope to identify all known bacteria. As stated above, any unidentifiable bacteria will require further study and prove that bacteria can grow at high altitudes. Phoenix /4/2006

14 Contact Information: Robert Lossing College of Arts and Sciences Nicole Dudley College of Engineering Nathan Lester College of Engineering Russell Morris College of Engineering Cory Smith College of Engineering Phoenix /4/2006