Sandwich freezing device for rapid freezing of viruses, bacteria, yeast, cultured cells and animal and human tissues in electron microscopy. (18th November 2020)

• Record Type: Journal Article
• Authors: Yamaguchi, Masashi; Taguchi, Masaki; Uematsu, Katsuyuki; Takahashi-Nakaguchi, Azusa; Sato-Okamoto, Michiyo; Chibana, Hiroji
• Is Part Of: Microscopy. Volume 70:Number 2(2021)
• Page Start: 215
• Page End: 223
• Publication Date: 2020-11-18
• Subjects: Cryo-electron microscopy -- freeze-substitution -- high pressure freezing -- rapid freezing -- sandwich freezing -- ultrastructure; Microscopy -- Periodicals; 502.825
• DOI: 10.1093/jmicro/dfaa049
• Languages: English
• ISSNs: 2050-5698

Abstract

Abstract: We have been using sandwich freezing of living yeast and bacteria followed by freeze-substitution for observing close-to-native ultrastructure of cells. Recently, sandwich freezing of glutaraldehyde-fixed cultured cells and human tissues have been found to give excellent preservation of ultrastructure of cells and tissues. These studies, however, have been conducted using a handmade sandwich freezing device and have been limited in a few laboratories. To spread the use of this method to other laboratories, we fabricated and commercialized a new sandwich freezing device. The new device is inexpensive, portable and sterilizable. It can be used to rapid-freeze viruses, bacteria, yeast, cultured cells and animal and human tissues to a depth of 0.2 mm if tissues are prefixed with glutaraldehyde. The commercial availability of this device will expand application of rapid freezing to wide range of biological materials.