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Schultz ER, Kelley KL, Paul AL, et al. A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy. Appl Plant Sci. 2013;1(8)doi: 10.3732/apps.1300034.
Schultz, E. R., Kelley, K. L., Paul, A. L., & Ferl, R. J. (2013). A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy. Applications in plant sciences, 1(8), . https://doi.org/10.3732/apps.1300034
Schultz, Eric R, et al. "A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy." Applications in plant sciences vol. 1,8 (2013). doi: https://doi.org/10.3732/apps.1300034
Schultz ER, Kelley KL, Paul AL, Ferl RJ. A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy. Appl Plant Sci. 2013 Jul 29;1(8). doi: 10.3732/apps.1300034. eCollection 2013 Aug. PMID: 25202579; PMCID: PMC4103452.
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Appl Plant Sci. 2013 Jul 29;1(8). doi: 10.3732/apps.1300034. eCollection 2013 Aug.

A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy.

Applications in plant sciences

Eric R Schultz, Karen L Kelley, Anna-Lisa Paul, Robert J Ferl

Affiliations

  1. Program in Plant Molecular and Cellular Biology, Horticultural Science Department, University of Florida, Gainesville, Florida 32611 USA.
  2. Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida 32610 USA.
  3. Program in Plant Molecular and Cellular Biology, Horticultural Science Department, University of Florida, Gainesville, Florida 32611 USA ; Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida 32610 USA ; Horticultural Sciences Department, University of Florida, Gainesville, Florida 32611 USA.

PMID: 25202579 PMCID: PMC4103452 DOI: 10.3732/apps.1300034

Abstract

PREMISE OF THE STUDY: In spaceflight experiments, tissues for morphologic study are fixed in 3% glutaraldehyde, while tissues for molecular study are fixed in RNAlater; thus, an experiment containing both study components requires multiple fixation strategies. The possibility of using RNAlater-fixed materials for standard SEM-based morphometric investigation was explored to expand the library of tissues available for analysis and maximize usage of samples returned from spaceflight, but these technologies have wide application to any situation where recovery of biological resources is limited. •

METHODS AND RESULTS: RNAlater-fixed samples were desalinated in distilled water, dehydrated through graded methanol, plunged into liquid ethane, and transferred to cryovials for freeze-substitution. Sample tissues were critical point dried, mounted, sputter-coated, and imaged. •

CONCLUSIONS: The protocol resulted in acceptable SEM images from RNAlater-fixed Arabidopsis thaliana tissue. The majority of the tissues remained intact, including general morphology and finer details such as root hairs and trichomes.

Keywords: Arabidopsis thaliana; RNAlater; SEM; scanning electron microscopy; spaceflight

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