In vivo veritas: electron cryotomography of cells

Implementation and performance evaluation of reconstruction algorithms on graphics processors

“…This increase is boosted by recent hardware developments, e.g. of the CCD cameras (more pixels and larger field of view), as well as the automatization of the data recording, both in electron tomography and single particle analysis (Carragher et al, 2000) (Plitzko et al, 2002) (Zhu et al, 2004). In order to cope with these increasing computational efforts, most of the software packages used in electron microscopy have implemented parallel reconstruction algorithms to be used on distributed computing systems (computing clusters) (Frank et al, 1996) (Sorzano et al, 2004).…”

Cryo-electron microscopy of vitreous sections of native biological cells and tissues

“…Typically, single macromolecules or macromolecular complexes like viruses, but in some cases even small bacteria or thin parts of other cells, are observed (Plitzko et al, 2002). Nonetheless, most cells and every tissues are not accessible to cryo-EM of thin films.…”

High-pressure freezing combined with in vivo-DAB-cytochemistry: A novel approach for studies of endocytic compartments

“…Previous studies proved the importance of a rapid vitrification of cells, e.g. by high-pressure freezing (HPF), for a detailed insight into architectures and organization of such complex and dynamic organelles (Bouchet-Marchis et al, 2008;Geerts et al, 2009;Hess et al, 2000;Knoll et al, 1987;Ladinsky et al, 1999;Leis et al, 2008;Lucić et al, 2008;Marsh, 2005;Marsh et al, 2001;Mogelsvang et al, 2004;Murk et al, 2003;Plitzko et al, 2002;Robinson and Karnovsky, 1991;Verkade, 2008). However, there exist limitations mainly concerning concurrent cytochemical analyses: in cryo-immobilized cells, the differentiation of endocytic compartments by a common, widely used cytochemical method, which localizes internalized peroxidase-labeled molecules (Graham and Karnovsky, 1966), is only possible with limitations.…”