CREATING HIGH QUALITY PLANT CUTICLE IMAGES WITH EXISTING METHODS AND NEW MICROSCOPIC SYSTEMS

Morphological information from plant cuticle is useful when analyzing growth and development of plant epidermal cells. Efficiently creating superior images for extracting morphological data from plant cuticle is now possible using existing methods coupled with improved microscopy systems. Our main aim is to obtain high quality images of cuticle or efficiently create large numbers of cuticle images to allow for more robust data analyses. Creating a larger dataset of detailed cuticle images will allow for more sophisticated and accurate analyses. We tested three different chemicals for maceration with three different stains on modern Red Maple (Acer rubrum) and Rocky Mountain Maple (Acer glabrum) cuticle samples. We macerated cuttings from leaves with either chromium trioxide, bleach, or chloral hydrate. Step dehydration from water to xylene was used for all samples, but one of three stains was used for each cuticle: PAS, toluidine blue, or propidium iodide. The cuticle were then placed on slides using Eukitt, a resin-based mounting medium. These slides were imaged using two different microscopes to compare the quality of images produced. The first microscope was a Hamamatsu Nanozoomer slide scanner, and the second microscope was a Zeiss Axioscan.Z1 slide scanner. Slide scanners create high-quality images of the cuticle in a relatively short amount of time, and images can provide information on epidermal cells and stomata on the adaxial side of the leaf. These methods for cuticle preparation can create higher quality images efficiently and could potentially be a new way in quantifying how external variables, such as climate, can affect the cell shape and epidermal development in leaves.