‘Cool ESEM’ - Imaging Ice-Containing Systems at Freezer Temperatures

Abstract Conventional scanning electron microscopy permits the use of cryogenic techniques, which has led to the examination of ice-containing samples (eg ice cream) at temperatures of -80°C and below. At these temperatures moderate etching of the ice crystals occurs which helps to identify the crystals in the surrounding glassy matrix. However, imaging at higher temperatures - equivalent to those at which most ice-containing systems are actually utilised - is impossible because of rapid sublimation. We have set out to develop a system which will enable us to examine ice crystals at temperatures of -20°C and below, corresponding much more closely to realistic conditions of use. In the environmental scanning electron microscope (ESEM), a gas can be maintained around the sample. Not only can this be used to maintain the sample in its native state, but the gas is actively involved in the imaging process through its role in signal amplification.