Invited Paper BP+AS-SuA3
3-D View into Cells by X-ray Nano-Tomography
Sunday, October 28, 2012, 4:40 pm, Room 23
| Session: |
Biomaterials Plenary - Bioimaging: In Vacuo, In Vitro, In Vivo |
| Presenter: |
G. Schneider, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany |
| Authors: |
G. Schneider, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany
P. Guttmann, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany
S. Werner, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany
K. Henzler, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany
S. Rehbein, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, Germany
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| Correspondent: |
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X-ray imaging offers a new 3-D view into cells. With its ability to penetrate whole hydrated cells it is ideally suited for pairing fluorescence light microscopy and nanoscale X-ray tomography. The HZB TXM at the undulator U41 provides a spectral resolution of 10.000 and a spatial resolution of 11 nm. For high resolution tomography, we adopted a tilt stage originally developed for electron tomography. The stage is able to tilt samples up to ± 80°. Such a large tilt of flat sample holders is impossible with TXM at bending magnet sources because they require a monochromator pinhole to be positioned close to the specimen. In our TXM, the holder geometry is no longer restricted to glass tubes.Conventional fluorescence images are diffraction-limited to ~200 nm, whereas current TXM achieve a ten-fold improvement in resolution. Since fluorescence and X-ray microscopy permit analysis of whole cells, it is possible to investigate the same cell in both microscopes by correlative microscopy. These correlative studies are ideally suited to X-ray microscopy because of its ability to image cells in 3D. In the talk, we present the cryo TXM and selected applications. In particular, we will show the internal structures of mammalian cells, i.e. plasma membrane,nuclear membrane, nuclear pores, nucleoli, endoplasmic reticulum, vesicles, lysosomes and mitochondria. It is now also possible to resolve internal organellar structures, such as mitochondrial cristae, the double nuclear membrane and lysosomal inclusions. In addition, we discuss ways towards 10 nm 3D imaging of cells. Keywords: X-ray microscopy, tomography, cell organelles, correlative microscopy References
1. S. Rehbein, S. Heim, P. Guttmann, S. Werner, G. Schneider, Phys. Rev. Lett. 103, (2009) 110801
2. G. Schneider, P. Guttmann, S. Heim, S. Rehbein, F. Mueller, K. Nagashima, J.B. Heymann, W.G. Müller, J.G. McNally, Nature Methods 7 (2010), 985-987
3. P. Guttmann, C. Bittencourt, S. Rehbein, P. Umek, X. Ke, G. Van Tendeloo, C. P. Ewels and G. Schneider, Nature Photonics 6 (2012), 25-29
4. G. Schneider, P. Guttmann, S. Rehbein, S. Werner, R. Follath, J. Struct. Biol. 177 (2012), 212-223