Using Silicon Polymer Impression Technique and Scanning Electron Microscopy to Measure Stomatal Aperture, Morphology, and Density

Background

The different techniques have been explored to study stomatal density and patterns in variety of plants. It can be broadly grouped into two classes including direct observation of fresh materials and the preparation of replicas, or castings (Gitz and Baker, 2009). These methods have included the use of clear nail varnish, which is a traditional method used to analyze stomata density, however, surfaces of leaves can be damaged by the solvent in the nail varnish. Here we use a non-destructive imprint of Arabidopsis thaliana leaves for stomata visualization, which can directly observe freshly collected materials and assess stomata density for sequential measurements. Additionally, it can simplify the observation and the measurement of stomatal density, and facilitate the analysis of several lines or different treatments in parallel.