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Correlative VIS-fluorescence and soft X-ray cryo-microscopy/tomography of adherent cells
| Content Provider | Semantic Scholar |
|---|---|
| Author | Hagen, Christoph Guttmann, Peter Klupp, Barbara G. Werner, Stephan Rehbein, Stefan Mettenleiter, Thomas C. Schneider, Gerd H. Grünewald, Kay |
| Copyright Year | 2012 |
| Abstract | Soft X-ray cryo-microscopy/tomography of vitreous samples is becoming a valuable tool in structural cell biology. Within the 'water-window' wavelength region (2.34-4.37nm), it provides absorption contrast images with high signal to noise ratio and resolution of a few tens of nanometer. Soft X-rays with wavelengths close to the K-absorption edge of oxygen penetrate biological samples with thicknesses in the micrometer range. Here, we report on the application of a recently established extension of the transmission soft X-ray cryo-microscope (HZB TXM) at the beamline U41-XM of the BESSY II electron storage ring by an in-column epi-fluorescence and reflected light cryo-microscope. We demonstrate the new capability for correlative fluorescence and soft X-ray cryo-microscopy/tomography of this instrument along a typical life science experimental approach - the correlation of a fluorophore-tagged protein (pUL34-GFP of pseudorabies virus, PrV, the nuclear membrane-anchored component of the nuclear egress complex of the Herpesviridae which interacts with viral pUL31) in PrV pUL34-GFP/pUL31 coexpressing mammalian cells, with virus-induced vesicular structures in the nucleus, expanding the nucleoplasmic reticulum. Taken together, our results demonstrate new possibilities to study the role of specific proteins in substructures of adherent cells, especially of the nucleus in toto, accessible to electron microscopy in thinned samples only. |
| Starting Page | 193 |
| Ending Page | 201 |
| Page Count | 9 |
| File Format | PDF HTM / HTML |
| DOI | 10.1016/j.jsb.2011.12.012 |
| PubMed reference number | 22210307 |
| Journal | Medline |
| Volume Number | 177 |
| Alternate Webpage(s) | http://xrm.phys.northwestern.edu/research/pdf_papers/2012/hagen_jsb_2012.pdf |
| Alternate Webpage(s) | https://doi.org/10.1016/j.jsb.2011.12.012 |
| Journal | Journal of structural biology |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
