An Overview of Cryo-Scanning Electron Microscopy Techniques for Plant Imaging.

Many research questions require the study of plant morphology, in particular cells and tissues, as close to their native context as possible and without physical deformations from some preparatory chemical reagents or sample drying. Cryo-scanning electron microscopy (cryoSEM) involves rapid freezing and maintenance of the sample at an ultra-low temperature for detailed surface imaging by a scanning electron beam. The data are useful for exploring tissue/cell morphogenesis, plus an additional cryofracture/cryoplaning/milling step gives information on air and water spaces as well as subcellular ultrastructure. This review gives an overview from sample preparation through to imaging and a detailed account of how this has been applied across diverse areas of plant research. Future directions and improvements to the technique are discussed.