title: Optical sectioning and 3D reconstructions as an alternative to scanning electron microscopy for analysis of cell shape.
abstract: OBJECTIVE
  Visualizing flower epidermal cells is often desirable for investigating the interaction between flowers and their pollinators, in addition to the broader range of ecological interactions in which flowers are involved. We developed a protocol for visualizing petal epidermal cells without the limitations of the commonly used method of scanning electron microscopy (SEM).
  
  
  METHODS
  Flower material was collected and fixed in glutaraldehyde, followed by dehydration in an ethanol series. Flowers were dissected to collect petals, and subjected to a Histo-Clear series to remove the cuticle. Material was then stained with aniline blue, mounted on microscope slides, and imaged using a compound fluorescence microscope to obtain optical sections that were reconstructed into a 3D image.
  
  
  RESULTS
  This optical sectioning method yielded high-quality images of the petal epidermal cells with virtually no damage to cells. Flowers were processed in larger batches than are possible using common SEM methods. Also, flower size was not a limiting factor as often observed in SEM studies. Flowers up to 5 cm in length were processed and mounted for visualization.
  
  
  CONCLUSIONS
  This method requires no special equipment for sample preparation prior to imaging and should be seen as an alternative method to SEM.
authors.affiliation: Department of Biology, University of Florida, 876 Newell Drive, Gainesville, Florida 32611 USA ; Florida Museum of Natural History, University of Florida, 1659 Museum Road, Gainesville, Florida 32611 USA.
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Landis
authors.affiliation: Florida Museum of Natural History, University of Florida, 1659 Museum Road, Gainesville, Florida 32611 USA.
authors.forename: Kayla L
authors.initials: KL
authors.lastname: Ventura
authors.affiliation: Department of Biology, University of Florida, 876 Newell Drive, Gainesville, Florida 32611 USA ; Florida Museum of Natural History, University of Florida, 1659 Museum Road, Gainesville, Florida 32611 USA.
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Soltis
authors.affiliation: Florida Museum of Natural History, University of Florida, 1659 Museum Road, Gainesville, Florida 32611 USA.
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Soltis
authors.affiliation: Department of Biology, University of Florida, 876 Newell Drive, Gainesville, Florida 32611 USA.
authors.forename: David G
authors.initials: DG
authors.lastname: Oppenheimer
keywords: confocal microscopy; fluorescence; morphology; optical sectioning; petal cells