Whole-mount Clearing and Staining of Arabidopsis Flower Organs and Siliques

J Vis Exp. 2018 Apr 12:(134):56441. doi: 10.3791/56441.

Abstract

Due to its formidable tools for molecular genetic studies, Arabidopsis thaliana is one of the most prominent model species in plant biology and, especially, in plant reproductive biology. However, plant morphological, anatomical, and ultrastructural analyses traditionally involve time-consuming embedding and sectioning procedures for bright field, scanning, and electron microscopy. Recent progress in confocal fluorescence microscopy, state-of-the-art 3-D computer-aided microscopic analyses, and the continuous refinement of molecular techniques to be used on minimally processed whole-mount specimens, has led to an increased demand for developing efficient and minimal sample processing techniques. In this protocol, we describe techniques for properly dissecting Arabidopsis flowers and siliques, basic clearing techniques, and some staining procedures for whole-mount observations of reproductive structures.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Arabidopsis / anatomy & histology*
  • Arabidopsis / ultrastructure
  • Flowers / ultrastructure
  • Microscopy, Confocal / methods*
  • Specimen Handling / methods*
  • Staining and Labeling / methods