Chapter 9 low-temperature electron microscopy: Techniques and protocols

Abstract

Low-temperature electron microscopy endeavors to provide “solidifi cation of a biological specimen by cooling with the aim of minimal displacement of its components through the use of low temperature as a physical fi xation strategy” (Steinbrecht and Zierold, Cryotechniques in biological electron microscopy. Springer- Verlag, Berlin, p 293, 1987). The intention is to maintain confi dence that the tissue observed retains the morphology and dimensions of the living material while also ensuring soluble cellular components are not displaced. As applied to both scanning and transmission electron microscopy, cryo-electron microscopy is a strategy whereby the application of low-temperature techniques are used to reduce or remove processing artifacts which are commonly encountered in more conventional room temperature electron microscopy techniques which rely heavily on chemical fi xation and heavy metal staining. Often, cryo-electron microscopy allows direct observation of specimens, which have not been stained or chemically fi xed.