title: A method for preparing spaceflight RNAlater-fixed Arabidopsis thaliana (Brassicaceae) tissue for scanning electron microscopy.
abstract: OBJECTIVE
  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. •
  
  
  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.
authors.affiliation: Program in Plant Molecular and Cellular Biology, Horticultural Science Department, University of Florida, Gainesville, Florida 32611 USA.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Schultz
authors.affiliation: Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida 32610 USA.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Kelley
authors.affiliation: Program in Plant Molecular and Cellular Biology, Horticultural Science Department, University of Florida, Gainesville, Florida 32611 USA.
authors.forename: Anna-Lisa
authors.initials: AL
authors.lastname: Paul
authors.affiliation: 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.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Ferl
keywords: Arabidopsis thaliana; RNAlater; SEM; scanning electron microscopy; spaceflight