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We have explored two ways to improve the spatial resolution and/or visibility of cellular ultrastructure in full field soft X-ray nanotomography at the Mistral beamline (ALBA light source). The first one consists on a new data collection method and processing framework to computationally extend the depth of field (XTEND) of a Fresnel zone plate objective lens based on focal series projections and deconvolution. Visual details of a 3D-reconstructed eukaryotic cell that is affected by depth of field artifacts in standard tomographic reconstruction are recovered. The second one consists on diminishing the missing wedge inherent to flat sample supports (rotation range ±70deg) by performing dual axis tomography. Both strategies clearly show a spatial resolution and visibility improvement on the reconstructed volumes while complying with radiation dose limitation. Examples of these methods on relevant biological applications will be presented.
Eva Pereiro,Joaquín Otón,Javier Conesa,Javier Chichon,Ana Perez-Berna,Andrea Sorrentino,Jose L. Carrascosa, andJose Maria Carazo
"Increasing spatial resolution in full-field soft x-ray nanotomography of cells (Conference Presentation)", Proc. SPIE 10389, X-Ray Nanoimaging: Instruments and Methods III, 1038903 (3 October 2017); https://doi.org/10.1117/12.2276706
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Eva Pereiro, Joaquín Otón, Javier Conesa, Javier Chichon, Ana Perez-Berna, Andrea Sorrentino, Jose L. Carrascosa, Jose Maria Carazo, "Increasing spatial resolution in full-field soft x-ray nanotomography of cells (Conference Presentation)," Proc. SPIE 10389, X-Ray Nanoimaging: Instruments and Methods III, 1038903 (3 October 2017); https://doi.org/10.1117/12.2276706