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X-ray diffraction microscopy on frozen hydrated specimens

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dc.contributor.advisor Jacobsen, Chris; en_US
dc.contributor.author Nelson, Johanna en_US
dc.contributor.other Department of Physics en_US
dc.date.accessioned 2012-05-15T18:05:19Z
dc.date.available 2012-05-15T18:05:19Z
dc.date.issued 1-Aug-10 en_US
dc.date.submitted Aug-10 en_US
dc.identifier Nelson_grad.sunysb_0771E_10213.pdf en_US
dc.identifier.uri http://hdl.handle.net/1951/55562
dc.description.abstract X-rays are excellent for imaging thick samples at high resolution because of their large penetration depth compared to electrons and their short wavelength relative to visible light. To image biological material, the absorption contrast of soft X-rays, especially between the carbon and oxygen K-shell absorption edges, can be utilized to give high contrast, high resolution images without the need for stains or labels. Because of radiation damage and the desire for high resolution tomography, live cell imaging is not feasible. However, cells can be frozen in vitrified ice, which reduces the effect of radiation damage while maintaining their natural hydrated state. X-ray diffraction microscopy (XDM) is an imaging technique which eliminates the limitations imposed by current focusing optics simply by removing them entirely. Far-field coherent diffraction intensity patterns are collected on a pixelated detector allowing every scattered photon to be collected within the limits of the detector's efficiency and physical size. An iterative computer algorithm is then used to invert the diffraction intensity into a real space image with both absorption and phase information. This technique transfers the emphasis away from fabrication and alignment of optics, and towards data processing. We have used this method to image a pair of freeze-dried, immunolabeled yeast cells to the highest resolution (13 nm) yet obtained for a whole eukaryotic cell. We discuss successes and challenges in working with frozen hydrated specimens and efforts aimed at high resolution imaging of vitrified eukaryotic cells in 3D. en_US
dc.description.sponsorship Stony Brook University Libraries. SBU Graduate School in Department of Physics. Lawrence Martin (Dean of Graduate School). en_US
dc.format Electronic Resource en_US
dc.language.iso en_US en_US
dc.publisher The Graduate School, Stony Brook University: Stony Brook, NY. en_US
dc.subject.lcsh Physics, Optics en_US
dc.subject.other coherent imaging, diffraction, x-ray microscopy, yeast en_US
dc.title X-ray diffraction microscopy on frozen hydrated specimens en_US
dc.type Dissertation en_US
dc.description.advisor Advisor(s): Chris Jacobsen. Committee Member(s): Peter Stephens; Marivi Fernandez Serra; Markus Seeliger. en_US
dc.mimetype Application/PDF en_US

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Nelson_grad.sunysb_0771E_10213.pdf 13.63Mb View/Open S2.mov 15.19Mb View/Open Movie of through-focus of reconstructed freeze-dried yeast wave field S1.mov 119.4Mb View/Open Movie of radiation damage visible in the autocorrelation of freeze-dried yeast

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