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

机译:线型X射线纳米断层扫描法测定细胞超微结构的三维质量密度图

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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 mu m-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. (C) 2015 The Authors. Published by Elsevier Inc.
机译:我们证明了绝对定量质量密度映射的三个方面的冷冻水合生物物质,各向同性分辨率为180 nm。作为生物系统的模型,我们在限制在微毛细管中的缓冲溶液中使用衣原体细胞。我们使用柱状X射线计算机断层摄影术对包括18微米直径毛细管的整个标本进行成像,从而直接提供了绝对质量密度的生物物质绝对测量值,不确定度约为6%。生成的图具有足够的对比度,可以将细胞与周围的冰区以及细胞内部不同密度的多个细胞器区分开。通过与染色的,树脂包埋的标本进行比较来识别细胞器,然后将其与已建立的透射电子显微镜结果进行比较。对于某些确定的细胞器,其元素组成的了解将其质量密度测量的不确定性降低至1%,其值与先前通过扫描透射电子显微镜测量的薄细胞切片中干重浓度的测量值一致。随着在不久的将来提高空间分辨率的前景,我们期望接近自然状态的生物样品中质量密度的非破坏性三维映射能力成为测量纳米级有机物堆积的一种有价值的方法。 (C)2015作者。由Elsevier Inc.发布

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