X-ray tomography of whole cells

Gros, Mark A. Le; McDermott, Gerry; Larabell, Carolyn A.

doi:10.1016/j.sbi.2005.08.008

Cited by 219 publications

(186 citation statements)

References 23 publications

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“…SXT uses the inherent contrast of fully hydrated specimens by imaging in the 'water window' (2.4 nm wavelength, 517 eV) where common biological elements such as carbon and nitrogen absorb X-rays and are therefore visible, whereas water is relatively transparent Larabell and Nugent, 2010;Le Gros et al, 2005;Patwardhan et al, 2014). Additionally, owing to the use of a thin-walled glass capillary tube to flash freeze and image the specimen, there is no visible radiation damage at the achievable resolution, and it is possible to collect projection image tilt series of 180°or greater allowing for a three-dimensional reconstruction with no distortions due to missing wedge artifacts as in electron microscopy (Larabell and Nugent, 2010;Le Gros et al, 2005Parkinson et al, 2008). Variations of this technique have been successfully applied towards the understanding of changes in cellular morphology due to disease and/or treatment (Chichón et al, 2012;Hanssen et al, 2012;Myllys et al, 2016;Pérez-Berná et al, 2016).…”

Section: Sxt Of Sickled Rbcsmentioning

confidence: 99%

Visualizing red blood cell sickling and the effects of inhibition of sphingosine kinase 1 using soft X-ray tomography

Darrow

Zhang

Cinquin

et al. 2016

Journal of Cell Science

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Sickle cell disease is a destructive genetic disorder characterized by the formation of fibrils of deoxygenated hemoglobin, leading to the red blood cell (RBC) morphology changes that underlie the clinical manifestations of this disease. Using cryogenic soft X-ray tomography (SXT), we characterized the morphology of sickled RBCs in terms of volume and the number of protrusions per cell. We were able to identify statistically a relationship between the number of protrusions and the volume of the cell, which is known to correlate to the severity of sickling. This structural polymorphism allows for the classification of the stages of the sickling process. Recent studies have shown that elevated sphingosine kinase 1 (Sphk1)-mediated sphingosine 1-phosphate production contributes to sickling. Here, we further demonstrate that compound 5C, an inhibitor of Sphk1, has anti-sickling properties. Additionally, the variation in cellular morphology upon treatment suggests that this drug acts to delay the sickling process. SXT is an effective tool that can be used to identify the morphology of the sickling process and assess the effectiveness of potential therapeutics.

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Section: Sxt Of Sickled Rbcsmentioning

confidence: 99%

Visualizing red blood cell sickling and the effects of inhibition of sphingosine kinase 1 using soft X-ray tomography

Darrow

Zhang

Cinquin

et al. 2016

Journal of Cell Science

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“…Phase contrast enhancement of X-ray images (Betz et al, 2007), while useable on unstained material, is much more useful for objects with distinct edges, rather than the softer gradients characteristic of embryos. Three-dimensional X-ray microscopy has been used to image subcellular structures (Larabell and Le Gros, 2004;Le Gros et al, 2005), but resolutions of tens of nanometers require a synchrotron source and a field of view too small to be of general utility to organismal developmental research.…”

Section: Introductionmentioning

confidence: 99%

MicroCT for developmental biology: A versatile tool for high‐contrast 3D imaging at histological resolutions

Metscher

2009

Developmental Dynamics

542

624

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Understanding developmental processes requires accurate visualization and parameterization of threedimensional embryos. Tomographic imaging methods offer automatically aligned and calibrated volumetric images, but the usefulness of X-ray CT imaging for developmental biology has been limited by the low inherent contrast of embryonic tissues. Here, I demonstrate simple staining methods that allow high-contrast imaging of embryonic tissues at histological resolutions using a commercial microCT system. Quantitative comparisons of images of chick embryos treated with different contrast agents show that three very simple methods using inorganic iodine and phosphotungstic acid produce overall contrast and differential tissue contrast for X-ray imaging at least as high as that obtained with osmium. The stains can be used after any common fixation and after storage in aqueous or alcoholic media, and on a wide variety of species. These methods establish microCT imaging as a useful tool for comparative developmental studies, embryo phenotyping, and quantitative modeling of development. Developmental Dynamics 238: 632-640, 2009.

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“…Microscopes that do not require focusing optics are classified as lensless projection imaging microscopes (PIMs) or diffraction imaging microscopes (DIMs). 91 As there are no lenses between the sample and the detector, higher resolution can be obtained, but it is limited by both the wavelength of the incident beam (synchrotron X-ray source) and by the strong decay of the diffraction intensity. 92 In biological samples, these techniques also allow good resolution (11- Recently, the same research team, have been able to measure 400 nm diameter gold nanocrystal in higher resolution.…”

Section: X-ray Microscopy (Xrm)mentioning

confidence: 99%

A General Perspective of the Characterization and Quantification of Nanoparticles: Imaging, Spectroscopic, and Separation Techniques

Lapresta-Fernández

Salinas-Castillo

Llana³

et al. 2014

Critical Reviews in Solid State and Materials Sciences

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This review gives an overview of the different techniques used to identify, characterize and quantify engineered nanoparticles (ENPs). The state-of-the-art of the field is summarized, the different characterization techniques have been grouped according to the information they can provide. In addition some selected applications are highlighted for each technique. The classification of the techniques has been carried out according to the main physical and chemical properties of the nanoparticles such as morphology, size, polydispersity characteristics, structural information and elemental composition. Microscopy techniques including optical, electron and X-ray microscopy and separation techniques with and without hyphenated detection systems are discussed. For each of these groups, a brief description of the techniques, the specific features and concepts, as well as several examples are described.

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X-ray tomography of whole cells

Cited by 219 publications

References 23 publications

Visualizing red blood cell sickling and the effects of inhibition of sphingosine kinase 1 using soft X-ray tomography

Visualizing red blood cell sickling and the effects of inhibition of sphingosine kinase 1 using soft X-ray tomography

MicroCT for developmental biology: A versatile tool for high‐contrast 3D imaging at histological resolutions

A General Perspective of the Characterization and Quantification of Nanoparticles: Imaging, Spectroscopic, and Separation Techniques

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