Electron microscopy of plant samples by using high-pressure freezing/freeze substitution method

Abstract

Sample fixation of electron microscopy is roughly classified into a conventional chemical fixation and freeze fixation method. Freeze fixation method has several advantages as compared to chemical fixation, for example, preventing outflow of cell substances and deformation of cell structures, high temporal resolution, holding antigens by immunoelectron microscopy. The high-pressure freezing method is suitable for plant samples which have large cells. To date, we have examined electron microscopic analysis of Arabidopsis thaliana and tobacco tissues and cultured cells as well as unicellular algae by high-pressure freezing/freeze substitution method. In this mini review, we will introduce basic application and technical findings of highpressure freezing/freeze substitution method, and discuss technical problems.