The Three Phases of Spaceflight Research – Prep, Flight and Analyses

By no means do these three categories fully encompass all of the intricacies of doing research in space, they do give us an easy way to break it down into discussable sections. Currently we are still in the Prep – or Pre-Launch phase of the APEX-03 experiment set to Launch on SpaceX-CRS 5 in the next week.

Today we will dive into some details on the prep phase –
After experimental methods and seed selection, the most important part of the Prep stage is planting the seeds that will be germinated on orbit. This is an important step because we need to place the seeds in their growth medium (in this case a .5x MS Phytagel nutrient gel media instead of soil) but we need them to not begin to grow until they have made it to the ISS, and have been installed in the hardware where they will grow. This time period can be anywhere between 2-4 weeks. We prepare these “dormant” plates of seeds by first sterilizing the seeds in ethanol to ready them for planting out on Petri plates containing sterile Phytagel growth medium. Step two is then test the sterilized seed batches to ensure they are indeed sterile, and that they have high germination rates when activated. Once we are confident in our seed quality, we can move into the next phase of preparation – Planting.

Planting for flight takes about half a day to set up, and then the remainder of the day to do the actual planting, providing you have the help of two additional lab members. To plant, the seeds are suspended in a small amount of sterile water to facilitate dispersing onto the surface of the nutrient gel in the Petri plate, and then the plates are immediately wrapped in black out cloth; keeping them in the dark inhibits germination until they are unwrapped on the space station. It is important that this all is done in a fully sterile environment and that from the time the water is added that only 10 minutes elapses before the plate is wrapped (take a look at the time-lapse video below – it shows Anna-Lisa Paul (left) and Jordan Callaham (right) planting in the laminar flow hood, and Eric Schultz (far right) outside the hood photographing and wrapping the plates.

[ Now you might ask – why is Eric photographing plates full of dormant seeds? I mean, they are just seeds, they all look exactly the same…The answer is that these are official “close-out photographs” required by NASA. Absolutely everything that goes to the space station is documented, cataloged and controlled. Because our plates are wrapped in black cloth when they are turned over, we need to provide documentation of what lies inside each black-wrapped packet. A non-flight example is shown below. ]

Plates containing 15-40 seeds are then stored in a refrigerator until the beginning of the experiment in space. We plant about twice as many plates as we will need for the flight (for the flight we need 27 plates, we plant 45-48). These extra plates serve as back up and test plates, so we can monitor them before the experiment is launched. And the flight experiment is comprised of more than just the set of plates that fly… 48 and 96 hours after the first set of plates are planted, second and third sets of plates are planted to serve as ground control and back-up (in case of launch delay.) These plantings require the same high level of attention that the flight plates receive – note, the ground control set is absolutely as important to the experiment as the set that flies!

Once all sets of plates are planted and stored in the refrigerator we monitor the test plates for any signs of problems before launch. As long as all plates look healthy and good to go, they will be turned over to NASA 48 hours prior to launch. These plates are then transported by NASA to the SpaceX Dragon capsule to be launched.

More to come soon about turn over operations and launch – as well as what is happening on flight!

UF Space Plants are headed back to Space!

A new series of UF Space Plants are soon on their way to the ISS, extending our science experiments on the ways that biology adapts to being in space. Our plants are part of the APEX-03 payload that will launch aboard the SpaceX-5 mission, set to launch at 2:31pm on December 16.

These experiments explore the deep reaches of plant root architectures that change during spaceflight. Results will provide new insight into how plants respond to extraterrestrial environments, which enables our understanding of how to better explore space and improves the research for growing food and producing oxygen on future space missions.

There will be eight individual seed lines flying, each one with a unique job in helping us determine plant responses to spaceflight. The plants will be grown on sterile petri plates, and harvested at 4, 8 and 11 days for later analysis once the samples return to Earth. We have a wonderful opportunity to image the plants while they are on the space station through the use of the LMM or Light Microscopy Module. This allows us to gather real-time data on the gene response to spaceflight environment.

Watch live on NASA TV at http://www.nasa.gov/multimedia/nasatv/index.html#.VIXDOzHF8mR . 

CARA at GRC

The GRC featured image of the day:  How Does This Garden Grow? click here

CARA – it’s a wrap.

Yesterday we traveled to KSC to retrieve the CARA samples that were harvested to on orbit, along with their comparable ground controls. The plants were harvested by Steve Swanson to KSC Fixation Tubes (KFTs) filled with “RNALater” – a solution that preserves the plants and genetic material in the state they were in on orbit for our later study on Earth.

Steve Swanson harvests plants on orbit to Kennedy Space Center Fixation Tubes - KFTs 

 Thursday one of the Payload Integration Specialists at KSC worked with us to unload the KFTs, and we brought the precious cargo home.  Thus ends the orbital portion of the CARA experiment!

One of 10 KFTs containing plants harvested on orbit, as returned to us at KSC

What’s next for CARA? Several months of laboratory analyses – imagine analyses from LMM and standard macro-photography, and the biochemical analyses of gene expression in the returned samples.

What’s next for us in the spaceflight realm? In two weeks we start in on structured preparation for our next flight experiment: TAGES-ISA. The Science Verification Test (SVT) for TAGES-ISA begins June 11th at KSC. The upcoming SVT will run a short version of our proposed flight experiment within the ISS Environmental Simulator (the ISSES chamber) and flight-comparable hardware (the VEGGIE unit). This test will – you guessed it – verify that the science of our proposed experiment is a good match for the hardware and likely environment it will see on the ISS.  
Later in the year we may be sending our biology to the suborbital realm as well – Stay tuned!

Video: Closing down LMM imaging

On Friday Steve Swanson pulled our petri plate out of the Light Microscopy Module and re-secured the LMM. The LMM will stay in active until its next deployment.

The imaging phase of the CARA experiment is now over. On Tuesday the remainder of the CARA plates will be collected and the plants on those plates will be harvested, preserved and frozen for return when the Dragon capsule makes its way back to earth.

ISS Video US Lab during LMM teleoperations

This video is still from the US lab, but the camera that was watching the LMM operations has been relocated to a general view of the US Laboratory Module. Interestingly is allows a change in perspective with regard to what direction is "up"

Gene expression images from ISS

The LMM has transferred some excellent images of plant roots on the ISS. Here is a white-light image showing a root tip growing on the surface of the agar medium. This image gives us precise location and morphology information. Below is a GFP gene expression image, showing the precise location of the cells within the root that are expressing this gene. The larger image at the bottom is a combined image, showing both the morphological location information as well as the gene expression data.

Steve Swanson taking the plate out of the LMM

Now that this imaging session is over, astronaut Steve Swanson spent a good bit of time opening up the rack and removing our plate of plants from the LMM.

Day of ISS imaging complete

Notebook page of image captures

The first 24-hr imaging session with the LMM was completed this morning! The UF team kept track of the imaging progress with old school notebook entries that allow correlation with data streams from the ISS. ISS images are being annotated and stored in preparation for transfer and analysis. The root image here shows the kind of high resolution images coming from the LMM.

White light image from the LMM of a root section

Video - Steve Swanson with the LMM

Telescience from GRC to the ISS

We scientists have always been impressed by the people that make science work. Especially in the business of space biology, there are so many skilled people working to make our science possible. For the next few days we will be here at Glenn Research Center in the Telescience Support Center. At this point, just after the LMM is enabled on orbit, the TSC is alive with a whole group of people communicating with the ISS, commanding the LMM to begin operations, and otherwise coordinating the transfer of instructions up and data down. So now these folks from Glenn join the folks from KSC and the folks from CASIS, all making good things happen for this unique kind of science.

Petri-Plants into the LMM on orbit!

Steve Swanson prepares a plate for the LMM

At 0645 EDT Astronaut Steve Swanson began the process to install the CARA imaging plate into the Light Microscopy Module LMM. Formally this begins the imaging section of the CARA project, which is known as Petri Plants. Actually PetriPlants is also the OpNom (Operational Nomenclature) for the CARA experiment. The documentation and the calls back and forth from the ISS refer to PetriPlants for ease of identification.

The UF team is at Glenn Research Center where the LMM is controlled from the Telescience Support Center TSC.

CARA is enabled by CASIS as part of ARK1

The ARK1 mission patch

CARA is a payload enabled by CASIS, the Center for the Advancement of Science in Space, as part of their initial science increment on the ISS. CASIS launch increments are referred to as Advancing Research Knowledge, or ARK. According to CASIS, "ARK1 marks the initial voyages of CASIS and its partner researchers."

Photos - CARA plates on the ISS

The CARA plates are all nicely positioned on the wall (floor?) of the ISS in the US Laboratory module. Here they will be exposed to light and the ambient ISS environment for their growth cycle. The more tightly arranged plates on the right (in this view) will grow in the light for the 12 days of the experiment. The more scattered 10 on the left were re-wrapped in black cloth and will provide a set of dark-grown controls for the experiment. Identical Ground Control sets are now growing in the ISSES Chamber (ISS Environment Simulator) at Kennedy Space Center (KSC).

On the adjoining wall of the module is the MELFI freezer, which can be identified by the circular doors that cover the actual freezer components.

U.S. Laboratory module

Astronaut Ops

The CARA experiment was successfully activated this morning at 7:04 a.m. EDT. The astronaut proceeded smoothly through the instructions, asked questions along the way, we answered, and the experiment began its growth phase on the ISS. Sounds simple, and in a way it is, but conversing with the astronaut is a little like a game of telephone…

It works like this: We scientists sit in the EMA with our Payload Integration Specialist. She talks with other payloads on a communications loop with the Payload Operations Director (POD), who in turn talks to the Payload Communications Manager (PAYCOM), and then PAYCOM conveys the message directly to the astronaut. It may sound complicated, but these are communications experts and it all works flawlessly.

What the astronaut sees when the payload is de-stowed from its cold transport bag is a set of plates wrapped in black cloth (1). Unwrapped, the seeds can be seen sown on the surface of the nutrient agar inside the Petri plate (2). In eight days, the seeds will look something like the plants in (3), and then on day 12 they will be harvested, looking much like the plants in (4).

Activation on the ISS 4/30/2014

This morning we are in the EMA (Experiment Monitoring Area), a mini-mission control room at KSC where our local payload integration specialist April Spinale and mission support engineer Joe Benjamin join the communications loops that connect with the ISS. They serve the ISS operations by answering questions and providing clarification on procedures. They also gather data as it comes down for the experiment records.

Within the EMA we have the closest real time connection with the astronaut. We can listen in on the audio loops and watch available video, and virtually participate in the experiment.

At 7:04 a.m. EDT astronaut Steve Swanson completed the activation of the experiment. He unwrapped the plates and put them under the lights for growth. Pictures were taken and we hope to get those downloaded soon.

Why Do We Study Space Biology?

Our science is driven by questions about what happens to terrestrial life when it leaves Earth. How successful can life be at moving away from Earth? What are the limits of life’s movements in the universe? More directly from a biological research perspective, what does it take to adapt to an extraterrestrial existence?

Think about it, the spaceflight environment is completely outside of the evolutionary experience of any terrestrial organism, so how do you “know” what to take out of your metabolic tool box to help you adapt? In space, plants know that they are in a place that lacks their customary cues, and as a consequence they engage a wide variety of responses to help navigate in this novel place.

We monitor plant responses by examining their changing patterns of gene expression – the breadcrumbs they drop as they explore different paths to acclimation.  We have been studying this for years now, using the modern tools of biotechnology and the spaceflight opportunities afforded by NASA programs.

Although our research focuses on the fundamental nature of how plants respond to novel environments, our space biology program is about more than just growing plants in space … it is also about understanding how to grow plants in small closed environments and optimizing protected habitats. These are issues that are important to us if we want to grow plants on a space station or on a long journey, but also if we want to grow plants in a greenhouse on Mars or the moon. Think of it as extreme protected agriculture! And indeed, the lessons we have learned and tools we have developed growing plants in a Martian-analog greenhouse in the arctic, and in small chambers on the shuttle and ISS, have direct application to protected agriculture here in Florida.

Update - Experiment Activation will happen on the ISS, Wednesday 4/30

The timeline for the Dragon capsule stay at the ISS is now established, setting in place the timeline for the CARA experiments. Astronaut Steve Swanson has set up the area in the ISS where the Petri plates with the plants will grow. Wednesday at 6 a.m EDT he will activate the experiment by pulling the plates out of cold stowage and exposing them to light to start the seeds germinating.

Here are images of the Petri plates, wrapped in black cloth to prevent seed germination, being loaded into the Double Cold Bag just before being placed in Dragon before launch. To activate the experiment Steve Swanson will open the cold bag, remove the temperature control cold blocks, then pull out the wrapped Petri plates. After they warm up he will unwrap them from the black cloth and mount them in the ISS.

Video - Our 2010 Experiments on the ISS

Over the coming weeks, our experiments aboard the ISS will be unpacked and activated by an ISS astronaut, and then manipulated from the ground via the Light Microscopy Module.

In 2010, we had the pleasure of working with astronaut Jeffrey Williams on the ISS. In the video above, you can watch Jeff work with one of our experiments. Using methods and actions similar to what goes on in labs here at the University of Florida, Jeff is harvesting plants, taking pictures, recording the data and placing the plants in fixation tubes to preserve them – in this case for their ride back to earth and UF.

The video shows him working in the ISS US Laboratory Module at a bench-like facility called the Maintenance Work Area - it has velcro and widget attachments to hold tools and samples in place. Jeff, like most all of the astronauts we have met, is a superb lab rat who cares deeply about conducting good science. (Actually, he looks pretty much like a typical graduate student, except that he is floating more than most of the students in our labs.)

The video is a little long and is green at the beginning. Fast forward to about 2:00 minutes to get to the action.

SpaceX Dragon Takes UF Plants To Space Station

A University of Florida science project that rocketed to the International Space Station on April 18 aboard the SpaceX-3 Dragon is focused, literally, on what it takes for biology to adapt and thrive in space.

The experiment by plant molecular biologists Robert Ferl and Anna-Lisa Paul uses small plants as models to understand cellular responses to spaceflight. This experiment, known as CARA, is a follow-up to research they conducted on the ISS in 2010 which found for the first time that roots display normal movements used to get around rocks and obstacles even when there is no gravity.

The movements, known as waving and skewing, were thought to be due to cellular responses to gravity pulling on roots as they sample their growing surface with touch, Paul said.

“But as the images from our experiment started to come down from the International Space Station in early 2010, it was clear that gravity was not required after all,” she said.

Based on the 2010 results, Paul and Ferl wanted to see if, in the absence of gravity, perhaps the plants were responding to another directional cue, such as the overhead light source integrated into the plant growth hardware. Such alternate signal processing by the plant cells would indicate that biology explores unique adaptive strategies when in novel circumstances.

“We are intrigued by the numerous light-sensing genes that are expressed specifically in roots in orbit, and the SpaceX-3 experiment further explored the role of these genes in orientation and cellular remodeling,” Paul said. “It is likely that light plays a more important role in root growth in microgravity than it does on Earth.”

And that has big implications for life somewhere other than Earth, Ferl said.

“This is telling us that life utilizes special, potentially unique signals to adapt to living off planet,” he said. “This has tremendous implications for the expansion of human existence to other worlds, but also richly informs us about the potential for plants to adapt to unusual environmental changes here on Earth.”

Ferl and Paul are working with NASA engineers at Glenn Research Center (GRC) in Ohio to adapt the Light Microscopy Module (LMM) for biological applications in order to focus the analysis on the cells that normally sense gravity in plants.

The LMM is a sophisticated fluorescent microscope facility housed on the ISS, with a counterpart at GRC, that has historically been used for physics experiments. The plants of the CARA experiment are grown in square petri plates on a nutrient agar matrix, and in this experiment simply attached to the wall of the US module of the ISS. During the course of the CARA experiment the plates are being photographed in place with a standard camera.

In addition, the plate containing plants engineered with fluorescent reporters are being examined with the LMM to evaluate spaceflight-associated changes in the cellular localization of selected genes.

An astronaut inserts the plate into a specially adapted holder on the on-orbit LMM, then Paul and Ferl work with the GRC engineers to control the use of the microscope telemetrically from the ground.

The experiment is sponsored as a research grant to Paul and Ferl by the Center for the Advancement of Science in Space (CASIS), the Florida-based national organization responsible for promoting science on the ISS.