Nanopositioning for the ESRF ID16A Nano-Imaging Beamline

Villar, F.; André, Lionel; Baker, R.; Bohic, Sylvain; Silva, Júlio César da; Guilloud, Cyril; Hignette, O.; Meyer, Jens; Pacureanu, Alexandra; Perez, Manuel; Salomé, Murielle; Linden, P. van der; Yang, Yiying; Cloetens, Peter

doi:10.1080/08940886.2018.1506234

Cited by 26 publications

(20 citation statements)

References 22 publications

(21 reference statements)

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“…The focus spot size was determined with a lithographic sample consisting of a 10 nm thick, 20 × 20 µm² square of nickel on a 500 nm thick SN membrane. The SN membranes holding the neurons were mounted in vacuum on a piezo nano-positioning stage with six short range actuators and regulated under the metrology of twelve capacitive sensors ( Villar et al, 2018 ). An ultra-long working distance optical microscope was used to bring the sample to the focal plane (depth-of-focus±3 µm) and to position the STED regions of interest in the X-ray beam (see section sample positioning below).…”

Section: Methodsmentioning

confidence: 99%

Correlating STED and synchrotron XRF nano-imaging unveils cosegregation of metals and cytoskeleton proteins in dendrites

Domart

Cloetens

Roudeau

et al. 2020

eLife

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Zinc and copper are involved in neuronal differentiation and synaptic plasticity but the molecular mechanisms behind these processes are still elusive due in part to the difficulty of imaging trace metals together with proteins at the synaptic level. We correlate stimulated-emission-depletion microscopy of proteins and synchrotron X-ray fluorescence imaging of trace metals, both performed with 40 nm spatial resolution, on primary rat hippocampal neurons. We reveal the co-localization at the nanoscale of zinc and tubulin in dendrites with a molecular ratio of about one zinc atom per tubulin-αβ dimer. We observe the co-segregation of copper and F-actin within the nano-architecture of dendritic protrusions. In addition, zinc chelation causes a decrease in the expression of cytoskeleton proteins in dendrites and spines. Overall, these results indicate new functions for zinc and copper in the modulation of the cytoskeleton morphology in dendrites, a mechanism associated to neuronal plasticity and memory formation.

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Section: Methodsmentioning

confidence: 99%

Correlating STED and synchrotron XRF nano-imaging unveils cosegregation of metals and cytoskeleton proteins in dendrites

Domart

Cloetens

Roudeau

et al. 2020

eLife

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“…The instrument developed at the ID16A beamline at the European Synchrotron (da Silva et al, 2017) can produce a highly brilliant and coherent X-ray probe focused to a spot below 30 nm, which we have used for holographic imaging. Another critical aspect is the accuracy of the rotational and translational stage movements to match the length scale of the expected resolution (< 100 nm); for this we employed active capacitive compensation to stabilize the sample (Villar et al, 2018). The sample itself must also remain stable throughout the measurements -a particular concern is heating and warping of the sample due to radiation absorption.…”

Section: Advances In Xnh Imaging Of Neural Circuitsmentioning

confidence: 99%

Dense neuronal reconstruction through X-ray holographic nano-tomography

Pacureanu

Maniates-Selvin

Kuan

et al. 2019

Preprint

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Elucidating the structure of neuronal networks provides a foundation for understanding how the nervous system processes information to generate behavior. Despite technological breakthroughs in visible light and electron microscopy, imaging dense nanometer-scale neuronal structures over millimeter-scale tissue volumes remains a challenge. Here, we demonstrate that X-ray holographic nano-tomography is capable of imaging large tissue volumes with sufficient resolution to disentangle dense neuronal circuitry in Drosophila melanogaster and mammalian central and peripheral nervous tissue. Furthermore, we show that automatic segmentation using convolutional neural networks enables rapid extraction of neuronal morphologies from these volumetric datasets. The technique we present allows rapid data collection and analysis of multiple specimens, and can be used correlatively with light microscopy and electron microscopy on the same samples. Thus, X-ray holographic nano-tomography provides a new avenue for discoveries in neuroscience and life sciences in general.

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“…XRF measurements were done using the scanning X-ray fluorescence microscopy setup of the ID16A Nano-Imaging beamline of the ESRF (Grenoble) 64 . A pair of multilayer-coated Kirkpatrick-Baez mirrors located 185 m downstream of the undulator source, was used to focus the incident X-rays at the energy of 17 keV down to a 23 nm (horizontal) × 37 nm (vertical) spot size.…”

Section: Methodsmentioning

confidence: 99%

Nano-imaging trace elements at organelle levels in substantia nigra overexpressing α-synuclein to model Parkinson’s disease

et al. 2020

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Sub-cellular trace element quantifications of nano-heterogeneities in brain tissues offer unprecedented ways to explore at elemental level the interplay between cellular compartments in neurodegenerative pathologies. We designed a quasi-correlative method for analytical nanoimaging of the substantia nigra, based on transmission electron microscopy and synchrotron X-ray fluorescence. It combines ultrastructural identifications of cellular compartments and trace element nanoimaging near detection limits, for increased signal-to-noise ratios. Elemental composition of different organelles is compared to cytoplasmic and nuclear compartments in dopaminergic neurons of rat substantia nigra. They exhibit 150-460 ppm of Fe, with P/Zn/Fe-rich nucleoli in a P/S-depleted nuclear matrix and Ca-rich rough endoplasmic reticula. Cytoplasm analysis displays sub-micron Fe/S-rich granules, including lipofuscin. Following AAV-mediated overexpression of α-synuclein protein associated with Parkinson's disease, these granules shift towards higher Fe concentrations. This effect advocates for metal (Fe) dyshomeostasis in discrete cytoplasmic regions, illustrating the use of this method to explore neuronal dysfunction in brain diseases.

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Nanopositioning for the ESRF ID16A Nano-Imaging Beamline

Cited by 26 publications

References 22 publications

Correlating STED and synchrotron XRF nano-imaging unveils cosegregation of metals and cytoskeleton proteins in dendrites

Correlating STED and synchrotron XRF nano-imaging unveils cosegregation of metals and cytoskeleton proteins in dendrites

Dense neuronal reconstruction through X-ray holographic nano-tomography

Nano-imaging trace elements at organelle levels in substantia nigra overexpressing α-synuclein to model Parkinson’s disease

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