Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Plants in space: A novel method for fixing plant tissue samples maximizes time, resources, and data

12.09.2013
At work on the International Space Station, researchers studying plant and cell growth in space encountered a challenge.

Imaging revealed interesting spaceflight-associated root morphologies. They needed to fix the tissues for further study back on Earth, but conventional fixation methods require separate fixatives depending on whether the sample is intended for molecular or morphological study.


This is a SEM image of Arabidopsis tissue processed using the new single fixation protocol developed by Schultz et al. Pictured is the adaxial leaf epidermis (3500x, scale bar = 8.57 ìm).

Credit: Image courtesy of Schultz et al.

If the scientists wanted to study how spaceflight affected patterns of gene expression central to morphological patterns of cell growth, they needed a fixation method that would allow them to study both perspectives.

Most scientists at work in the laboratory rely on protocols that have been developed without the need for restrictions on the amount of space, time, or reagents they use. For scientists conducting experiments in spaceflight, time and resources are strictly regulated and limited, and researchers must know in advance which protocols will maximize the usefulness of the data they collect.

University of Florida professors Anna-Lisa Paul and Robert Ferl and colleagues are collaborating with the National Aeronautics and Space Administration (NASA) to understand plant growth and development in spaceflight. Along with lead author and graduate student Eric Schultz, they have developed a single fixation protocol for use in space that allows plant material to be used for multiple experimental applications. Their new protocol for sample preparation was tested on Arabidopsis tissues harvested on the International Space Station and is described in the August issue of Applications in Plant Sciences (available for free viewing at http://www.bioone.org/doi/pdf/10.3732/apps.1300034).

Because of limitations in astronaut crew time and orbital resources, previous spaceflight fixation protocols were designated as either molecular or morphological, due to the separate fixatives required for each application. Tissues for morphologic study were fixed in 3% glutaraldehyde (or a similar solution), and tissues for molecular study were fixed in the tissue storage reagent RNAlater. RNAlater has not commonly been used as a morphologic fixative, as it can produce unclear images with high background staining.

The new method developed by Schultz et al. puts RNAlater-fixed samples through a desalination process to return them to a fresh-like state, and then uses low-temperature scanning electron microscopy (cryo-SEM) to preserve tissues for imaging. Because few laboratories have access to the necessary equipment for cryo-SEM, the authors tested and developed a protocol that emulates cryo-SEM using standard SEM equipment and, importantly, that results in minimal tissue damage.

Although it was developed to address specific constraints for spaceflight experiments, Paul notes that their new method is broadly applicable. "There are a lot of situations where biologists want to collect samples in extreme situations. In our case—a space vehicle orbiting the Earth."

The new protocol maximizes the amount of data obtained from a single sample and allows for the concomitant examination of both molecular and morphological features. Using a single fixation protocol, direct comparisons between changes in morphology and altered gene expression can be made. Such an analysis not only makes full use of samples and replicates but also enables a robust analysis of the relationship between heredity and development. "Putting two tools together, it is powerful to look at the morphology in conjunction with the genes that are being expressed," says Paul.

The new protocol boasts low costs and high accessibility, and has wide application to any situation where recovery of biological resources is limited. Notably, this includes researchers collecting and preserving samples in the field, where space for materials is at a premium. "In places where sampling is limited, difficult, or expensive, the use of preservatives allows for more routes to analysis," notes Ferl.

Applications in Plant Sciences (APPS) is a monthly, peer-reviewed, open access journal focusing on new tools, technologies, and protocols in all areas of the plant sciences. It is published by the Botanical Society of America (http://www.botany.org), a non-profit membership society with a mission to promote botany, the field of basic science dealing with the study and inquiry into the form, function, development, diversity, reproduction, evolution, and uses of plants and their interactions within the biosphere. The first issue of APPS published in January 2013; APPS is available as part of BioOne's Open Access collection (http://www.bioone.org/loi/apps).

For further information, please contact the APPS staff at apps@botany.org.

Beth Parada | EurekAlert!
Further information:
http://www.botany.org

More articles from Life Sciences:

nachricht New risk factors for anxiety disorders
24.02.2017 | Julius-Maximilians-Universität Würzburg

nachricht Stingless bees have their nests protected by soldiers
24.02.2017 | Johannes Gutenberg-Universität Mainz

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Breakthrough with a chain of gold atoms

In the field of nanoscience, an international team of physicists with participants from Konstanz has achieved a breakthrough in understanding heat transport

In the field of nanoscience, an international team of physicists with participants from Konstanz has achieved a breakthrough in understanding heat transport

Im Focus: DNA repair: a new letter in the cell alphabet

Results reveal how discoveries may be hidden in scientific “blind spots”

Cells need to repair damaged DNA in our genes to prevent the development of cancer and other diseases. Our cells therefore activate and send “repair-proteins”...

Im Focus: Dresdner scientists print tomorrow’s world

The Fraunhofer IWS Dresden and Technische Universität Dresden inaugurated their jointly operated Center for Additive Manufacturing Dresden (AMCD) with a festive ceremony on February 7, 2017. Scientists from various disciplines perform research on materials, additive manufacturing processes and innovative technologies, which build up components in a layer by layer process. This technology opens up new horizons for component design and combinations of functions. For example during fabrication, electrical conductors and sensors are already able to be additively manufactured into components. They provide information about stress conditions of a product during operation.

The 3D-printing technology, or additive manufacturing as it is often called, has long made the step out of scientific research laboratories into industrial...

Im Focus: Mimicking nature's cellular architectures via 3-D printing

Research offers new level of control over the structure of 3-D printed materials

Nature does amazing things with limited design materials. Grass, for example, can support its own weight, resist strong wind loads, and recover after being...

Im Focus: Three Magnetic States for Each Hole

Nanometer-scale magnetic perforated grids could create new possibilities for computing. Together with international colleagues, scientists from the Helmholtz Zentrum Dresden-Rossendorf (HZDR) have shown how a cobalt grid can be reliably programmed at room temperature. In addition they discovered that for every hole ("antidot") three magnetic states can be configured. The results have been published in the journal "Scientific Reports".

Physicist Dr. Rantej Bali from the HZDR, together with scientists from Singapore and Australia, designed a special grid structure in a thin layer of cobalt in...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

Booth and panel discussion – The Lindau Nobel Laureate Meetings at the AAAS 2017 Annual Meeting

13.02.2017 | Event News

Complex Loading versus Hidden Reserves

10.02.2017 | Event News

International Conference on Crystal Growth in Freiburg

09.02.2017 | Event News

 
Latest News

Stingless bees have their nests protected by soldiers

24.02.2017 | Life Sciences

New risk factors for anxiety disorders

24.02.2017 | Life Sciences

MWC 2017: 5G Capital Berlin

24.02.2017 | Trade Fair News

VideoLinks
B2B-VideoLinks
More VideoLinks >>>