STEM tomography of high-pressure frozen cell monolayers

Abstract

We are investigating structures of the Golgi apparatus in relation to protein kinase D2 expression in pancreatic cells (see also Krisp et al, this issue) and membrane envelopment of capsids during viral morphogenesis in the cytoplasm of the host cells (see also Landwehr et al., this issue). Both processes are highly dynamic and for a better understanding, a three-dimensional description is preferable. For this purpose we have optimized the existing preparation and imaging protocols for electron tomography with the following goals: -Fast immobilization of a physiologically defined state. -Good preservation and good visibility of cellular components, especially of membranes. - Imaging of sections as thick as possible, so that a good three-dimensional impression of the dynamic processes can be achieved.