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Scanning transmission electron microscopy through-focal tilt-series on biological specimens

Abstract : Since scanning transmission electron microscopy can produce high signal-to-noise ratio bright-field images of thick (>= 500 nm) specimens, this tool is emerging as the method of choice to study thick biological samples via tomographic approaches. However, in a convergent-beam configuration, the depth of field is limited because only a thin portion of the specimen (from a few nanometres to tens of nanometres depending on the convergence angle) can be imaged in focus. A method known as through-focal imaging enables recovery of the full depth of information by combining images acquired at different levels of focus. In this work, we compare tomographic reconstruction with the through-focal tilt-series approach (a multifocal series of images per tilt angle) with reconstruction with the classic tilt-series acquisition scheme (one single-focus image per tilt angle). We visualised the base of the flagellum in the protist Trypanosoma brucei via an acquisition and image-processing method tailored to obtain quantitative and qualitative descriptors of reconstruction volumes. Reconstructions using through-focal imaging contained more contrast and more details for thick (>= 500 nm) biological samples. (C) 2015 Elsevier Ltd. All rights reserved.
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Submitted on : Monday, April 11, 2016 - 5:43:10 PM
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Sylvain Trépout, Cédric Messaoudi, Sylvie Perrot, Philippe Bastin, Sergio Marco. Scanning transmission electron microscopy through-focal tilt-series on biological specimens. Micron, 2015, 77, pp.9-15. ⟨10.1016/j.micron.2015.05.015⟩. ⟨pasteur-01301207⟩



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