Sandwich freezing device for rapid freezing of viruses, bacteria, yeast, cultured cells and animal and human tissues in electron microscopy

Author:

Yamaguchi Masashi1ORCID,Taguchi Masaki2,Uematsu Katsuyuki2,Takahashi-Nakaguchi Azusa1,Sato-Okamoto Michiyo1,Chibana Hiroji1

Affiliation:

1. Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan

2. Marine Works Japan, Ltd., 3-54-1 Oppamahigashi, Yokosuka 237-0063, Japan

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.

Publisher

Oxford University Press (OUP)

Subject

Radiology, Nuclear Medicine and imaging,Instrumentation,Structural Biology

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