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Femtosecond diffractive imaging of biological cells
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular biophysics.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular biophysics.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular biophysics.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular biophysics.
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2010 (English)In: Journal of Physics B: Atomic, Molecular and Optical Physics, ISSN 0953-4075, E-ISSN 1361-6455, Vol. 43, no 19, 194015- p.Article in journal (Refereed) Published
Abstract [en]

In a flash diffraction experiment, a short and extremely intense x-ray pulse illuminates the sample to obtain a diffraction pattern before the onset of significant radiation damage. The over-sampled diffraction pattern permits phase retrieval by iterative phasing methods. Flash diffractive imaging was first demonstrated on an inorganic test object (Chapman et al 2006 Nat. Phys. 2 839-43). We report here experiments on biological systems where individual cells were imaged, using single, 10-15 fs soft x-ray pulses at 13.5 nm wavelength from the FLASH free-electron laser in Hamburg. Simulations show that the pulse heated the sample to about 160 000 K but not before an interpretable diffraction pattern could be obtained. The reconstructed projection images return the structures of the intact cells. The simulations suggest that the average displacement of ions and atoms in the hottest surface layers remained below 3 angstrom during the pulse.

Place, publisher, year, edition, pages
2010. Vol. 43, no 19, 194015- p.
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-147259DOI: 10.1088/0953-4075/43/19/194015ISI: 000281958100016OAI: oai:DiVA.org:uu-147259DiVA: diva2:400246
Available from: 2011-02-25 Created: 2011-02-24 Last updated: 2017-12-11Bibliographically approved
In thesis
1. Flash Diffractive Imaging in Three Dimensions
Open this publication in new window or tab >>Flash Diffractive Imaging in Three Dimensions
2012 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

During the last years we have seen the birth of free-electron lasers, a new type of light source ten billion times brighter than syncrotrons and able to produce pulses only a few femtoseconds long. One of the main motivations for building these multi-million dollar machines was the prospect of imaging biological samples such as proteins and viruses in 3D without the need for crystallization or staining. This thesis contains some of the first biological results from free-electron lasers.

These results include 2D images, both of whole cells and of the giant mimivirus and also con- tains a 3D density map of the mimivirus assembled from diffraction patterns from many virus particles. These are important proof-of-concept experiments but they also mark the point where free-electron lasers start to produce biologically relevant results. The most noteworthy of these results is the unexpectedly non-uniform density distribution of the internals of the mimivirus.

We also present Hawk, the only open-source software toolkit for analysing single particle diffraction data. The Uppsala-developed program suite supports a wide range fo algorithms and takes advantage of Graphics Processing Units which makes it very computationally efficient.

Last, the problem introduced by structural variability in samples is discussed. This includes a description of the problem and how it can be overcome, and also how it could be turned into an advantage that allows us to image samples in all of their conformational states.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2012. 68 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 960
Keyword
X-ray, diffraction, mimivirus, three dimensional, phase retrieval, EMC, manifold embedding, CXI, FEL, free-electron laser, single particle
National Category
Biophysics
Research subject
Physics with specialization in Biophysics
Identifiers
urn:nbn:se:uu:diva-179643 (URN)978-91-554-8439-2 (ISBN)
Public defence
2012-10-05, B22, BMC, Husargatan 3, Uppsala, 09:15 (English)
Opponent
Supervisors
Available from: 2012-09-14 Created: 2012-08-21 Last updated: 2013-01-22Bibliographically approved

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Seibert, M. MarvinSvenda, MartinEkeberg, TomasMaia, Filipe R. N. C.Timneanu, NicusorCaleman, CarlAndersson, IngerHajdu, Janos

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Seibert, M. MarvinSvenda, MartinEkeberg, TomasMaia, Filipe R. N. C.Timneanu, NicusorCaleman, CarlAndersson, IngerHajdu, Janos
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Molecular biophysics
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Journal of Physics B: Atomic, Molecular and Optical Physics
Biological Sciences

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