SpaceX CRS-2 Launch!

In a few hours, the Gilroy lab BRIC-17 team will be at the OSB II (Operations Support Building 2) at KSC to watch the launch. OSB is about three and a half miles away from the launch pad, so we hope to have a good view. We’ll be bussed there from the visitor center early in the morning. Following a launch briefing we will wait for the launch at an outside viewing area, camera in hand!

SpaceX CRS-2 Logo

Unfortunately, we will be enjoying the launch after pulling an all-nighter in order to integrate our ground controls into the BRICs with the help of NASA hardware experts. The timing of our experiment is such that our science has to be integrated into the space hardware from 3-5am Eastern time. Our flight BRICs were prepared early Wednesday morning; because the ground control is offset by 2 days we have to make a repeat set of BRICs now (early Friday morning).

What if the launch is cancelled? The scrub schedule is such that the next launch attempt will occur just under 24hr later. If there are two scrubs, our experiment needs to be replaced with a fresh setup. Therefore, if the launch is cancelled both Friday and Saturday, our two BRICs will be pulled from Dragon. The ground controls we set up tonight will then become the flight BRICs and loaded into the Dragon in time for the third launch attempt Sunday morning, and so Sunday morning between 3-5am Eastern time we will have to set up two more BRICs for the new ground controls.

Simon and Won-Gyu in front of the NASA headquarters at KSC.

Tomorrow’s launch chance is at 80%. Everyone, please do us a huge favor and keep your fingers crossed for an on-time departure! (That means you too, Sis!)

In other news, yesterday (Thursday Feb 28) Simon participated in two news events.  The first was an interview with Jerry Hume , a reporter with the local Fox channel 13. We were told that while the transcript will be available online, the video will not because it will only be accessible to cable subscribers. It will air between 5am and noon Friday.

Jerry Hume interviews Simon about our BRIC-17 experiment.

The second news event was a pre-launch NASA press conference to discuss the science going up on SpaceX 2. The location of the press conference was in the NASA auditorium in the KSC press center, and for me it was a thrill to actually be sitting in a place that previously I’ve only seen on TV and online. There were five folks in front of the cameras, plus Josh the moderator:

• Julie Robinson, program scientist, International Space Station
• Simon Gilroy, BRIC-17 Lead Investigator,
University of Wisconsin
• Marshall Porterfield, division director, Life and Physical Sciences
NASA Headquarters
• Michael Johnson, Chief Technical Officer, NanoRacks
• Michael Roberts, Research Scientist, CASIS

Simon behind the desk at the NASA pre-launch press conference.

Simon did a great job communicating information specifically about our BRIC-17 experiment and speaking more generally about problems that living organisms have in space. He also fielded a number of questions from reporters ranging  from social media bloggers to tech reporters from the mainstream media. The video of this press conference is not yet available online, when it is I will update with a link.

BRIC-17 Integration

With the official SpaceX launch planned for Friday March 1 at 10:10am, the NASA BRIC team has scheduled our pre-launch meetings and integration. While still in Madison, Simon participated in a “readiness review” meeting with NASA on Friday, going through the checklist of everything that will need to occur before flight. We have the “green light” to go ahead with our BRIC experiment. We also have a good idea of how timing of events this week will occur.

The three members of our Gilroy Lab team (Sarah, Simon, and Won-Gyu) arrived in Florida Sunday night and moved into a condo in Cocoa Beach, which will be our home for at least a week. Bright and early Monday morning we went to the badging station at Kennedy Space Center to get our temporary passes onto the base. After that, we were able to cross the security checkpoint and enter the Space Station Processing Facility (SSPF).

Cocoa Beach, Florida

Last year for our initial tests at KSC (the SVT and PVT), our lab was located at the Space Life Science (SLS) lab. However, it made more sense to consolidate all of the NASA research into the SSPF building.  The SSPF has extra space now that the space station is built, and many rooms have been repurposed in to labs and offices. We were concerned that the move would cause some difficulty for our flight integration, but so far the new space is as good if not better as the SLS lab.

Our lab suite is split in two with the other half in use by a group of Canadian researchers who will send up an experiment to study microflow on the ISS. When we arrived at our lab Monday morning, we ventured across the dividing line. Strangely, the Canadian half is really very much like our American side, just with a different accent. In fact the corridor where our lab is has many other labs on it, each with a research team working hard on setting up for either our flight or the next SpaceX launch in September. This is a pretty exciting place to be at the moment!

Entry to the SSPF.

Our office in the SSPF is upstairs in a large partitioned room. The BRIC-17 teams are on one side, with three desks for UW-Madison and another three for the University of Florida team. In the same room are other groups flying experiments on SpaceX2, including the microfluidics group from Canada and the Japanese space agency JAXA team lead by Sachiko Yano who will be flying stem cells.

Simon at his desk in the SSPF.

Tonight (Tuesday Feb 26) we have a pre-integration meeting with the NASA folks who will be packing our experiment into the space hardware (the petri dish fixation unit, PDFU). The meeting is scheduled for 10:30pm because integration of BRIC-17 science into the PDFU will occur from 1am to 3am. We have been advised to bring plenty of food, because neither the cafes in the SSPF nor any nearby restaurants will be open.

Following integration, our BRICs will be stored in the refrigerator. Our seeds will be on nutrient gel, so it will be important that they be kept cool to prevent germination until their arrival in microgravity. Then, on Wednesday afternoon the loaded BRICs will be placed into a cold bag and moved to the SpaceX loading area. They should be on board the SpaceX Dragon capsule on Thursday in time for the Friday morning launch. The Dragon will dock with the space station on Saturday March 2, and our experiment will run from March 2 until the astronaut adds the RNAlater fixative to the samples on March 9 at 7:40pm. At 8pm the following day, the astronaut will place our BRICs into the freezer (MELFI) until landing back on Earth on March 25. The BRICs will be in the freezer at KSC until we can return to de-integrate and retrieve our samples in early April.

Won-Gyu planting seeds onto nutrient gel on Tuesday afternoon to prepare our ISS experiment.

The ground control will be offset by 2 days, so that KSC can get the on-board conditions (temperature) in order to program the growth chamber (the Orbital Environmental Simulator, OES) to mimic the space station conditions. Thus the schedule for set-up and running the ground control will be identical to the flown experiment, except everything will occur 2 days later. So we will have another all-nighter starting Thursday night to finish ground control science integration early Friday morning. We should be done in time to go watch the launch at 10:10am.

SpaceX and NASA have also established a “scrub schedule.” That is, if SpaceX does not launch on March 1 10:10am for some reason, then next launch attempt will be just slightly less than 24hr later (i.e. March 2 9:47am; if that second attempt is scrubbed then the next try will be March 3 9:24am, etc.). Our experiment is good for 2 scrubs; if two scrubs occur our ground control becomes our flight BRICs. Therefore, we will need to re-do our setup every other day until launch. Let’s hope all goes well and SpaceX launches on schedule March 1!

Botany experiment will try out zero gravity aboard space station

Last month, Simon was interviewed about our space station experiment by Chris Barncard of University Communications. I think Chris has put out an entertaining write-up, have a look and see if you agree!

University of Wisconsin’s news report about the Gilroy Lab experiment

University of Wisconsin – Madison

TOAST Launch Date: March 1, 2013

We’ve just received confirmation that the SpaceX 2 (SpX-2) launch is now officially scheduled for March 1, 2013. NASA has entered it on its Consolidated Launch Schedule web site.

NASA’s Launch Schedule

SpX-2 is the second NASA-contracted payload inside SpaceX’s Dragon capsule. The first was a space station resupply mission in October 2012, and the third is slated for October 2013. SpX-2 will carry the two BRIC-17 experiments up to the space station, including our Arabidopsis from the Gilroy Lab at the University of Wisconsin – Madison and plant cell cultures from the Paul Lab at the University of Florida – Gainesville.

Last month, the BRIC-17 teams completed their Payload Verification Test (PVT) at the Kennedy Space Center in Florida. This was a “dry run” of the experiment as it would be set up and run for the actual launch. Given our germination problems during the SVT (Science Verification Test) in October, we wanted to be sure that our revised method worked perfectly.

We tested three ways of setting up our seeds in the 60mm petri plate.

The method with perfect success during the PVT, and so the method we will use for our space flown experiment, was placing the seeds directly on Phytagel. Phytagel is a nutrient-containing “jello-like” material that we pour as a warm liquid into the bottom of the petri plate. Once it is cool, it forms a firm surface on which we place surface-sterilized seeds.

For our experiment, we will put 64 seeds in an 8×8 grid on the Phytagel surface in a total of ten 60mm petri plates. This is the science that will be loaded into the space hardware (PDFU inside BRICs) by NASA specialists and stored at 4 degrees C until launch. The cool temperature will prevent germination until the start of the experiment in microgravity. For launch, our BRICs will be placed in a cold bag to maintain the cool temperature while on board the Dragon capsule en route to the ISS. Once on the ISS, the astronauts will place our BRICs at ambient temperature and the experiment begins. This warmer temperature (20-22 deg C) will allow germination and seedling growth in microgravity.

Following 8 days of growth, the astronauts will inject RNAlater solution to fix our microgravity-grown seedlings, then the BRIC will be placed in the freezer. After our experiment returns to Earth, the NASA team will open our BRIC to retrieve our samples (deintegration). The plants should look similar to our plants grown during the PVT, shown below.

Photograph of a 60mm petri plate after deintegration from the PDFU space hardware, showing etiolated seedlings grown on Phytagel.

The seedlings are “etiolated,” which means that they are grown in the dark. Dark-grown Arabidopsis has a long shoot (hypocotyl) and relatively short root. The leaves do not expand or turn green. The lead scientist on our project, Won-Gyu Choi, isolated RNA from the shoot and the root of our PVT plants. The RNA contains transcripts of the genes that are expressed in the plant at the end of the 8 days of growth; the RNA expression provides a snapshot of how the plant was regulating its gene expression in response to its environment. For example, from Earth-based research we know what genes show enhanced expression during low oxygen stress in plants. We will compare the expression of these anoxia-induced genes in the space-grown material to the control plants we grow in the Orbial Environmental Simulator at the Kennedy Space Center.

An image of a gel containing good quality and abundant quantity of RNA isolated from seedlings grown for the PVT.

This morning, our PVT results were carefully examined by the NASA team in charge of the BRIC projects, and any risk factors thoroughly considered. This is called the Phase C Review. Following extensive review, the BRIC-17 projects were given NASA’s official stamp of approval for flight on SpX-2. Yay!

Now, all we can do is wait until the end of February 2013, when the Gilroy Lab team will again travel to Kennedy Space Center to prepare our samples to be loaded into the Dragon capusule for the March 1 launch of SpaceX-2.

OCD with the SVT

This past summer, the Gilroy Lab team traveled to Kennedy Space Center (KSC) at the Space Life Science (SLS) lab in Florida for our Science Verification Test (SVT). I returned to KSC two weeks later to process and ship our samples.

The results from our SVT were nothing like we expected! I’ll explain more about this in a moment.

The SVT is the first run for our experiment inside the actual space hardware called a Petri Dish Fixation Unit (PDFU). This hardware with our experiment inside is “integrated” by NASA engineers and staff during the SVT (i.e. the whole equipment is put together as if it was ready for being sent to the International Space Station). This is so that everyone knows what will happen when the time comes to do it for real a few days before launch. Then, during the SVT the experiment is run in the SLS lab inside a special growth chamber called an Orbital Environmental Stimulator (OES) that mimics conditions on the space station. At the end of the experiment, our samples are then “deintegrated,” that is, the PDFU is taken apart and the petri dish containing our plants is removed. Finally, our samples are processed and shipped to the Gilroy Lab at UW-Madison for analysis.

The OES (Orbital Environmental Simulator) growth chamber at NASA’s Space Life Science lab.

It is a requirement that the SVT practice run be completed so all of us can be confident that the experiment as planned has the highest probability of success.

Assembling the PDFU’s is no small task. NASA’s Susan is the integration team leader, and has an amazing ability to stay focused for hours doing the detailed work required to assemble PDFU after PDFU. During assembly, Susan calls out each component that she puts in place and Jennifer, the NASA quality control expert, takes note to confirm all is done correctly. Everything needs to be sterile, so all PDFU parts have been autoclaved and the assembly work is done in a sterile hood. This video shows Susan assembling a PDFU for a different NASA experiment from John Kiss’ lab; the process for our experiment is similar.

http://www.youtube.com/watch?v=riNaH4busxc

Once the PDFUs are put together with our science inside, a stack of 5 PDFU are placed in the aluminum BRIC canister. The sixth space in the BRIC is for a temperature data logger called a HOBO. Next, another team of NASA specialists conducts a leak test by placing the assembled BRIC in a pressure chamber. All of our PDFUs passed the leak test!

George conducts the pressure leak test on our PDFUs.

For the actual launch, our two freshly assembled BRICs will be placed in the fridge until transfer in cold storage to the SpaceX launch vehicle. Once the BRICs are on the space station, they will be brought to room temperature (22 deg C) and then a day later an astronaut will actuate the injection of growth media to start our experiment. For our SVT, this temperature regime and injection timing is replicated on the ground at the lab.

Then, after 8 days of growth, a fixative is added to our petri plates via another injection. After sitting in fixative at room temperature for a day, the BRICs are frozen until they can be returned to Earth, taken apart, and the samples shipped to the Gilroy Lab for analysis.

Susan removing our samples from a PDFU.

So, why then were our SVT results nothing like we had expected? We were disappointed to observe that when we got our SVT petri plates back from KSC that very few of the seeds had germinated.

In the weeks that followed, we did as much testing as we could in the Gilroy Lab to troubleshoot this germination problem. Was the PDFU plastic releasing a volatile chemical inhibiting germination? We tried activated charcoal cloth with the idea that it would absorb volatiles, however, the addition of charcoal cloth did not help. Was it the glue used to attach the filter paper to the dish? We confirmed that with enough drying time, the presence of glue had no impact on germination rate.

Was the germination problem due to the filter paper that the seeds were attached to? We tried different batches of filter paper to see if we could improve germination. It turned out that the old batch of paper we initially used was much better than the new. What could be different? We washed the filter paper before autoclaving, and that seemed to help somewhat. Then, we treated the paper with an ion solution before autoclaving; this treatment also improved our seed germination rate.

Our backup plan is to use petri plates containing a layer of nutrient solution in a gel called Phytagel, and place our seeds on top of that. This experimental design with wet gel is not as good as dry filter paper because the seeds will imbibe prior to launch. Thus, our dry seeds will start taking up moisture when they are placed on the gel during integration and not at the point when the astronauts inject growth media. However, the canisters holding our petri plates will be in cold storage in the fridge, therefore seeding growth will not be significant until the experiment starts at room temperature on the space station. This is a good alternate plan, and in fact the nutrient gel method has been used in past plant biology BRIC experiments with success.

Liz, Jose, and Janet discussing details during our SVT.

We’ve got another test run before actual launch, called the Payload Verification Test (PVT) in October. We’re hoping that we’ve sorted out any possible problems and that our PVT will be a success!

How to put TOAST in a BRIC?

Welcome to the inaugural post of the Gilroy Lab blog. In the coming weeks, we will be chronicling the trials and tribulations involved in putting a plant biology experiment on the space station. I’m Sarah and I’ll be the main contributor, with help from my boss Simon. The lead scientist working on the project is Won-Gyu, a post-doc in the Gilroy Lab. Please check the “People” link above for more information about us.

We were thrilled that our grant proposal was funded by NASA earlier this year! We’ll be studying how plants grow in microgravity and observing the impact that the space environment has on plant gene expression. Currently, our experiment is scheduled to fly in December up to the International Space Station (ISS) on the SpaceX Dragon/F9 spacecraft. Our experiment is called: Test Of Arabidopsis Space Transcriptome, aka TOAST.

The hardware that will contain our plants was developed by NASA engineers and has been flown numerous times with great success. This hardware is called Biological Research In Canister, or BRIC for short. Hence, we will be putting TOAST in a BRIC. I will go into more detail about our idea and how we are testing it in upcoming blog posts. However, if you want information right away about our experiment and the hardware flown, then please check the “Experiment” link above.

I am looking forward to sharing our experience with you. If you have any questions, please post a comment and I will reply as soon as I can. I hope to post here at least once a week, so check back often if you are curious as to how things are going.

The International Space Station.