Three-dimensional mass density mapping of cellular ultrastructure by ptychographic X-ray nanotomography - PubMed (original) (raw)

. 2015 Dec;192(3):461-469.

doi: 10.1016/j.jsb.2015.10.008. Epub 2015 Oct 22.

Barbora Malkova 2, Mirko Holler 2, Manuel Guizar-Sicairos 2, Enju Lima 3, Valerie Panneels 2, Gaia Pigino 4, Anne Greet Bittermann 5, Larissa Wettstein 2, Takashi Tomizaki 2, Oliver Bunk 2, Gebhard Schertler 2, Takashi Ishikawa 2, Roger Wepf 5, Andreas Menzel 2

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Three-dimensional mass density mapping of cellular ultrastructure by ptychographic X-ray nanotomography

Ana Diaz et al. J Struct Biol. 2015 Dec.

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Abstract

We demonstrate absolute quantitative mass density mapping in three dimensions of frozen-hydrated biological matter with an isotropic resolution of 180 nm. As model for a biological system we use Chlamydomonas cells in buffer solution confined in a microcapillary. We use ptychographic X-ray computed tomography to image the entire specimen, including the 18 μm-diameter capillary, thereby providing directly an absolute mass density measurement of biological matter with an uncertainty of about 6%. The resulting maps have sufficient contrast to distinguish cells from the surrounding ice and several organelles of different densities inside the cells. Organelles are identified by comparison with a stained, resin-embedded specimen, which can be compared with established transmission electron microscopy results. For some identified organelles, the knowledge of their elemental composition reduces the uncertainty of their mass density measurement down to 1% with values consistent with previous measurements of dry weight concentrations in thin cellular sections by scanning transmission electron microscopy. With prospects of improving the spatial resolution in the near future, we expect that the capability of non-destructive three-dimensional mapping of mass density in biological samples close to their native state becomes a valuable method for measuring the packing of organic matter on the nanoscale.

Keywords: Chlamydomonas; Ptychography; X-ray microscopy.

Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

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