X-ray microscopy is increasingly used in biology, but in most cases only in a qualitative way. We present here a 3D correlative cryo X-ray microscopy approach suited for the quantification of molar concentrations and structure in native samples at nanometer scale. The multimodal approach combines X-ray fluorescence and X-ray holographic nanotomography on "thick" frozen-hydrated cells. The quantitativeness of the X-ray fluorescence reconstruction is improved by estimating the self-attenuation from the 3D holography reconstruction. Applied to complex macrophage cells, we extract the quantification of major and minor elements heavier than phosphorus, as well as the density, in the different organelles. The intracellular landscape shows remarkable elemental differences. This novel analytical microscopy approach will be of particular interest to investigate complex biological and chemical systems in their native environment.
Cryo-nanoimaging of single human macrophage cells. 3D structural and chemical quantification / Gramaccioni, Chiara; Yang, Yang; Pacureanu, Alexandra; Vigano, Nicola; Procopio, Alessandra; Valenti, Piera; Rosa, Luigi; Berlutti, Francesca; Bohic, Sylvain; Cloetens, Peter. - In: ANALYTICAL CHEMISTRY. - ISSN 0003-2700. - 92:7(2020), pp. 4814-4819. [10.1021/acs.analchem.9b04096]
Cryo-nanoimaging of single human macrophage cells. 3D structural and chemical quantification
Valenti, Piera;Rosa, Luigi;Berlutti, Francesca;
2020
Abstract
X-ray microscopy is increasingly used in biology, but in most cases only in a qualitative way. We present here a 3D correlative cryo X-ray microscopy approach suited for the quantification of molar concentrations and structure in native samples at nanometer scale. The multimodal approach combines X-ray fluorescence and X-ray holographic nanotomography on "thick" frozen-hydrated cells. The quantitativeness of the X-ray fluorescence reconstruction is improved by estimating the self-attenuation from the 3D holography reconstruction. Applied to complex macrophage cells, we extract the quantification of major and minor elements heavier than phosphorus, as well as the density, in the different organelles. The intracellular landscape shows remarkable elemental differences. This novel analytical microscopy approach will be of particular interest to investigate complex biological and chemical systems in their native environment.File | Dimensione | Formato | |
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