Eureka

How it feels when an experiment isn't working - and then Eureka!

Video of a Night Blooming Cereus, taken by Dr. Paul. While this is one post that has little to do with space biology, it is an interesting example of terrestrial plant biology. However, it does connect with why we work with plant in space... we are primarily interested in how plants perceive and respond to their environment, and what molecular tools they use in the process. How do Arabidopsis roots know which direction to grow in the absence of gravity in the spaceflight environment? How does the night-blooming Epiphyllum Hookeri know to wait until around 10:00pm to open to attract its favorite pollinators? Both plants signals cued by light to help guide patterns of growth and development in interesting environments.

Keep your eyes on SpaceX CRS 10, aka Spx-10 this weekend! https://spaceflightnow.com/tag/spacex-10/

This launch of APEX-04 takes UF Spaceplants back to pad 39A where the moon shots and space shuttles rocketed into space.  Its been a few years since our experiments last left that pad, on STS131

Stay tuned to the twitter feed for any last minute launch updates.

The Conversation: Taking plants off planet – how do they grow in zero gravity?

Dr. Ferl and Dr. Paul recently had the opportunity to participate in The Conversation....check out the article they wrote...

http://theconversation.com/taking-plants-off-planet-how-do-they-grow-in-zero-gravity-45032

Flight Video

Check out a video of our most recent flight on the NASA Weightless Wonder VI.  In the video you can see how we work to complete research in even the most interesting environments.  

Parabolic Flight Campaign 2015 - Mission and Current Status

In a previous post, we discussed what parabolic flights are and why they are used.  But why are we using them this time?  We are flying our custom imaging systems (FLEX) to observe how various genes behave in zero gravity in three distinct ways.  These methods are either contained within FLEX imager itself or are external experiments designed to complement and verify the data collected within the imager.

We have two FLEX imager units, named Rocky and Bullwinkle.  One unit will be operational on the ground, and the other will fly on the C-9 parabolic aircraft.  This experimental design allows parallel data collection so we know that when we compare flight to ground, our differences are solely in what is happening during flight.

Part one of our experiment is green fluorescent protein (GFP) imaging.  This system will take pictures of GFP in our plants by using blue LEDs to excite the GFP and filters to see where the GFP is located.  We have already tagged GFP to genes of interest, and using this system will allow us to watch where these genes are active how they behave over the course of the parabolic flight.

Part one is supported by a floor harvest.  During the course of a parabolic flight, we can do 10 parabolas before we run out of airspace and need to turn around.  During these turns, we are able to open our Arabidopsis plates, take all the plants off of the plates, and put them into a solution that "freezes" (chemically preserves) the plants.  We can to this at each of the turns (after 10, 20, 30, and 40 parabolas).  For this campaign, we are harvesting the same plant "breeds" (lines) we are imaging in part one to verify that the change in GFP we may see through the camera is correct.

Part two of our experiment uses a FLIR thermal imaging camera.  We are comparing two different lines of Arabidopsis where one of the lines cools itself better than the other.  We want to see how the leaf temperature of these lines differ in zero gravity, since air can move differently in zero g.

Part three is an experiment designed to see what happens to plants at the beginning of one of these flights.  Using a Kennedy Space Center Fixation Tube (KFT), we are going to preserve plants at various early points in the flight to separate the plant responses to hyper gravity and zero gravity.

Currently, we are in Building 993 at Ellington Air Field, awaiting the final go-ahead that we are ready to fly.  Rocky and Bullwinkle are ready to go, and so are all of our plants.  More updates soon! - Eric Schultz

Parabolic Flights – More than just the “Vomit Comet”

We have posted online the past several weeks that we are currently gearing up for a new parabolic flight campaign in Houston, TX.  But what is involved in a parabolic flight campaign?  What is a parabolic flight?  And why do we even do them?  Hopefully, this brief overview will answer these questions, or at least provide an initial framework for where to go next.

A parabolic flight campaign is the entire mission associated with the parabolic flights.  For this campaign, that involved a pre-Test Readiness Review (TRR) on Friday, June 5^th, followed by the actual TRR the following Monday (as well as loading the plane and installing hardware), four consecutive flight days (Tuesday-Friday), unloading the aircraft on Friday, with a backup day on Saturday.  Since we study plants, that means all of our planting must be done beforehand, and must be done so that each day, we can have the same age of plants.  For this campaign, that means planting dormant plates and activating them in a staggered pattern, so that we can have (for example) 8 day old plants on Tuesday, Wednesday, Thursday, and Friday.  Since we will be in Texas, and not close to UF, we also have to arrange travel, lodging, meals, and set up a portable workstation and laboratory to do the work we need to do while we are there.  A lot of planning goes into one of these campaigns, usually starting about three months before we depart.  But it is this planning that makes these campaigns possible—and the more planning, the more successful it is too.

Image credit: "Zero gravity flight trajectory C9-565" by NASA - C-9B Flight Trajectory, NASA Reduced Gravity Research Program. Licensed under Public Domain via Wikimedia Commons - http://commons.wikimedia.org/wiki/File:Zero_gravity_flight_trajectory_C9-565.jpg#/media/File:Zero_gravity_flight_trajectory_C9-565.jpg

A parabolic flight uses commercial aircraft to achieve true zero gravity by adjusting its angle of flight.  The diagram (above) shows one parabola and how the plane is able to achieve this altered gravity state.  In our case, we will fly forty parabolas in one flight, for a campaign total of 160 parabolas.  We depart and arrive at Ellington Airport (formerly Ellington Field) in Houston, TX.  What about the nickname, “Vomit Comet?”  Though it is not the official name of the aircraft (that would be “Weightless Wonder”), it does accurately reflect the relatively high rate of motion sickness induced by this kind of a flight pattern.  Anti-motion sickness medication is usually administered to all flyers prior to each flight, and from experience, they really do work.  The flight is very smooth also.  Save for an abrupt end to each of the zero gravity portions, the rest of the flight is more akin to a boat’s motion than a roller coaster.  Weightlessness is truly a unique feeling.

We use parabolic flight to see rapid responses to zero gravity, hyper gravity, and changes in gravity in general.  NASA has used parabolic flight in the past to train astronauts, and many other institutions and organizations have used them for testing their systems and techniques in zero gravity, prior to true spaceflight.  Being a plant space biology laboratory, we are particularly interested in plant response to gravity.  Plants have not evolved to respond specifically to zero gravity, and thus in order to adapt to this new environment; they must engage previously-existing pathways.  What parabolic flight allows us is to look at what happens first—the plant’s initial response to a change in gravity, whether that is from normal 1g to 2g (beginning of the parabola), 2g to 0g (middle), or 0g to 2g (end).  Spaceflight allows us to look at adaptation to a prolonged zero gravity environment, which is extremely valuable.  Combined with parabolic flight, we can generate a more complete picture of how plants respond to changes in gravity. 

Here is Dr. Paul explaining a little more about what it is like to ride on the "Vomit Comet".

What do interceptor-class jets and plants have in common? A lot more than you might think…

In April of 2013, our lab performed an experiment with Starfighters Aerospace, located in Kennedy Space Center.  This company uses Lockheed F-104s to create the same extreme g-forces felt during suborbital missions.  These craft are able to fly at Mach 2.2 and are able to climb to altitudes over 90,000 feet.  For more information, visit http://www.starfighters.net/

But what do Starfighters have to do with plants?  Our experiment looked at plant transcriptome response to suborbital flight profiles.  The basic question we asked was, “What would plants feel during a suborbital flight?”  We were also asking if an untrained civilian (represented here by our two PIs, Dr. Rob Ferl and Dr. Anna-Lisa Paul) could be tasked to perform certain functions during the flight, such as taking various measurements and harvesting plants in a special chemical fixation unit designed at Kennedy Space Center called a KFT.  We also flew more plants in the two separate cargo holds, one of which was pressurized.  We are currently analyzing the data from microarrays, which tell us the transcriptional profile of the plants from KFTs, and will soon be analyzing the differences in atmospheric pressures, in addition to the g-forces felt on this kind of flight.

The flight itself consisted of a high-speed takeoff, three lateral turns, a high-speed, low-altitude run, followed by a 90-degree vertical climb.  The F-104 then turned upside-down, resulting in about 10 seconds of zero gravity, then began the high-g descent.  This profile is similar to the forces that will be felt on suborbital craft, the only difference being the amount of time in zero gravity.  The entire process was repeated two to three times for each flight.  The video below is a montage of the two flights from that day that represents the flight profile and some of the activities performed onboard.

Dragon is back! Samples on their way...

Dragon on deck

Imaging phase is over - awaiting sample return

It is with a small sense of relief, and a large sense of gratitude, that we now close down the LMM image gathering segment of the APEX-03 experiment. The crew at GRC have been fantastic and we deeply appreciate their efforts. Now we return our focus to the flight operations that center with our colleagues at KSC. Having successfully managed the flight operations and the ground control operations, the team at KSC is back in our forefront, monitoring the packing and stowing of the APEX samples for return on Dragon.

We will all be watching for a safe departure of Dragon from the ISS and a successful splash down.

Third and final APEX harvest and imaging off to a great start

This morning at about 0230 EST CDR Butch Wilmore began the setup and actual final harvest of the APEX-03 experiment on the ISS. The harvest went extremely well. CDR Wilmore's expertise and attention to detail, plus his familiarity with the APEX experiments, all contributed to a fast and efficient harvest that was amazing to see.

The last imaging plate was installed into the LMM to complete the harvest activity, and the imaging team at GRC is and will be hard at work for the next 28 hours collecting the final sets of on orbit images of Arabidopsis gene expression.

Outstanding stuff!

Second LMM Imaging Session: Video of astronaut closing up the LMM

Here is a part of the process that we have not been able to show previously. This video shows the front window of the glovebox that holds the stage of the LMM. You can see our petri plate sitting on the stage (it is sitting flat in this perspective, presenting a side view with the white tape wrap visible, just to the left of the glove).Later in the video you can see CDR Wilmore swing the glovebox and LMM back into the FIR rack in preparation for closing up the rack panels. Once this is complete the rack is powered on and the team from GRC takes over to command the images.

and busier...

Well, we survived the first set (flight and ground) of LMM operations – one set down, two to go. We need to keep long and unusual hours, and we trade off shifts to allow the other to get some rest. Being from Florida, driving to the base at 0200 in the snow is a new reality… As said earlier – we are busy, but it is the price of being able to do exciting science in an amazing environment.

Below is a composite image of one of the roots we collected with the orbital LMM this week. It shows a section of root expressing two different fluorescent gene reporters, one tracked with GFP and the other tracked with RFP. The white light image is stacked with the red and green images to produce this map off gene expression and adaptation to spaceflight.

Also below is a time lapse movie of astronaut Cdr. Butch Wilmore working with our experiment,captured with a cell phone from one of the monitors. We have already posted some snippets of his operations, but the time-lapse is pretty fun to watch.
Some of the corresponding Ground Control operations can be seen below as well; Allison Mjoen of KSC is the pseudonaut.

and finally, a small look at the outside of GRC building 333, which houses the LMM GIU and the Telescience Support Center.

Imaging is moving along

Busy!

The harvest on orbit went fine, and that started in motion a long list of data gathering activities that have taken all the time of the crew here at GRC. To give you some idea of what we look at, here is an animation of what the product looks like. The root morphology is in white, with a halo of lighter white water around it, and the specific gene expression areas are in  green. Once we get all the LMM images taken, post processing turns those raw images into image stacks and volumes that look like this. Know what genes are expressed in what areas of the root is just the information that we are trying to develop. It will really help our understanding of gravity related signalling in the root tips and help us better define the adaptation of plants to spaceflight.

ISS Video- CDR with science sampling

Here is a short video of CDR Barry Willmore working the APEX harvest. Butch has proven to be a careful and attentive bench scientist and the harvests have proceeded without a hitch. Nice work!

UF Spaceplants on their way to the LMM

The first UF Spaceplants harvest of APEX-03 is happening this morning. The UF team arrived at GRC at about 2:30AM EST to get set up watching the ISS videos and listening to the comms links. Its cold and snowy in Cleveland and the entrance to GRC actually looks quite special under these conditions.

Nice night/morning to arrive at Glenn

ISS CDR Butch Wilmore is in now in the middle of the harvest procedure. He is at the MWA, the maintenance work area, which is all set up as the ISS lab bench for this experiment. Here is a short video of the video of the early procedures. In this video you can also see a portion of the crew planning document on the left hand side.

Operations at Glenn Research Center (GRC) are off to a great start. We spent the day running through the LMM (Light Microscopy Module) data collection protocols that we will be using for our flight experiment on Monday.  We are using our standard 10 cm square petri plates, but in addition to plants, the plates are also decorated with a series of fluorescent markers that we will use for calibration.

Step one is to take a reference photo of the plate. For this test run, the below reference photo was taken with my cell phone – on orbit the reference photo of the LMM plate will be taken by the astronaut with a sophisticated SLR camera.

Step two is to attach the petri plate to the LMM sample holder by means of Velcro tabs. This stabilizes the plate, and on orbit prevents it from floating off the surface.

Step three is the insertion of the sample holder into the Ground Imaging Unit (GIU) of the LMM, and (in this case) draping the microscope in black cloth to prevent stray light from interfering with fluorescent imaging.

Step four is to walk back from the High Bay where the LMM is housed, into the comfort of the Telescience Center.

From here,  we and a team of engineers “drive” the LMM; all operations – finding plant parts in the field of view, focusing, and collecting images – is done telemetrically. Although the ‘scope is only about 200 yards away, we conduct the experiment exactly as if it were, say, 200 miles above the surface of the earth… both the ground  and space station the role of navigators (and sometimes backseat drivers) as we peruse what the LMM reveals about how our plants are responding to their environment. Pretty amazing stuff.

Starting LMM operations at Glenn

Today we are working with the GRC crew that tends the LMM on the ISS. Today we spin up operations with the LMM and its ground unit the GIU, with the goal of establishing the procedures that we will use starting early Monday morning on the ISS. We are working out exposure values, magnifications and imaging locations so that all will be ready for the imaging on orbit.

Glenn Research Center is great place located in a really pretty part of the valleys and woods in this part of Cleveland. Of course it is still winter here so for us from UF it is a big change compared to Florida weather :)

The entrance into GRC, early this morning.

Ground Controls all good

Ground controls off and running!

Ground Controls starting today at KSC

Today the ground control experiments are getting started. The ground controls are a perfectly matched set of plates going into the ISSES chambers at KSC. These chambers are well matched to ISS local environment conditions such as temperature and CO2. And there is a veggie plant growth unit that is identical to the one on the ISS. This allows us to have a matched set of controls for comparison to the flight samples

Views of the ISSES Chambers and the insertion of APEX plates

ISS Operations done for the day

At this point the APEX experiment appears to be off to a great start. No reports of anomalies with the plates or the hardware. We are hoping for some downloaded HD video later in the day.

Closing out this activation process now sets the timeline for the rest of the time that Dragon is docked to the ISS. Our next operation is scheduled for the 26th, when the first harvest plates will be harvested and the Imaging Plate will be transferred to the Light Microscopy Module for GFP. In the meantime we will be monitoring growth conditions, checking with the KSC operations crew, and generally getting ready to start the on orbit analysis phase.