A deep 3D view into cells using cryo-scanning transmission electron tomography

Date:
23
Tuesday
January
2018
Lecture / Seminar
Time: 10:00-11:00
Location: Nella and Leon Benoziyo Building for Biological Sciences
Lecturer: Dr. Shron G. Wolf
Organizer: Department of Biomolecular Sciences
Details: Department of Chemical Research Support- WIS
Abstract: CryoSTEM tomography (CSTET) is a novel combination [1-5] of two well-established ... Read more CryoSTEM tomography (CSTET) is a novel combination [1-5] of two well-established techniques: cryo-electron tomography (CET) and scanning transmission electron microscopy (STEM). Cryogenic fixation allows for the most faithful preservation of biological macromolecules, tissues and cells. By preserving the cryogenic temperature and maintaining a contamination-free environment inside the electron microscope, CET for such cryo-preserved specimens have resulted in major advances in their characterization [6]. However, due to technical constraints, sample thickness amenable to viewing by CET is limited (up to ~400 nm). STEM provides a possibility for relieving these limitations because contrast and signal are produced incoherently. The natural limit for sample thickness is extended three-fold for CSTET, meaning that whole vitrified mammalian cells can be imaged at ~3-4 nm resolution for regions up to ~1 μm in thickness. For spreading cells, many inter- organelle contacts and morphologies can be studied. In addition, CSTET combined with Energy Dispersive X-ray Spectroscopy (EDX) allows for on-the-spot chemical characterization of cell contents. References 1. Wolf, S.G., L. Houben, and M. Elbaum, Cryo-scanning transmission electron tomography of vitrified cells. Nature Methods, 2014. 11(4): p. 423-428. 2. Wolf, S.G., P. Rez, and M. Elbaum, Phosphorus detection in vitrified bacteria by cryo- STEM annular dark-field analysis. J Microsc, 2015. 260(2): p. 227-33. 3. Elbaum, M., S.G. Wolf, and L. Houben, Cryo-scanning transmission electron tomography of biological cells. MRS Bulletin, 2016. 41(07): p. 542-548. 4. Wolf, S.G., E. Shimoni, M. Elbaum, and L. Houben, STEM Tomography in Biology, in Cellular Imaging, Electron Tomography and Related Techniques, E.G. Hanssen, Editor. 2017, Springer, in press. 5. Wolf, S.G., Y. Mutsafi, T. Dadosh, T. Ilani, Z. Lansky, B. Horowitz, S. Rubin, M. Elbaum, and D. Fass, 3D visualization of mitochondrial solid-phase calcium stores in whole cells. Elife, 2017. 6: e29929. 6. Beck, M. and W. Baumeister, Cryo-Electron Tomography: Can it Reveal the Molecular Sociology of Cells in Atomic Detail? Trends Cell Biol, 2016. 26(11): p. 825-837.
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