Dataset Information

Iron-specific Signal Separation from within Heavy Metal Stained Biological Samples Using X-Ray Microtomography with Polychromatic Source and Energy-Integrating Detectors.

ABSTRACT: Biological samples are frequently stained with heavy metals in preparation for examining the macro, micro and ultra-structure using X-ray microtomography and electron microscopy. A single X-ray microtomography scan reveals detailed 3D structure based on staining density, yet it lacks both material composition and functional information. Using a commercially available polychromatic X-ray source, energy integrating detectors and a two-scan configuration labelled by their energy- "High" and "Low", we demonstrate how a specific element, here shown with iron, can be detected from a mixture with other heavy metals. With proper selection of scan configuration, achieving strong overlap of source characteristic emission lines and iron K-edge absorption, iron absorption was enhanced enabling K-edge imaging. Specifically, iron images were obtained by scatter plot material analysis, after selecting specific regions within scatter plots generated from the "High" and "Low" scans. Using this method, we identified iron rich regions associated with an iron staining reaction that marks the nodes of Ranvier along nerve axons within mouse spinal roots, also stained with osmium metal commonly used for electron microscopy.

SUBMITTER: Katchalski T

PROVIDER: S-EPMC5953933 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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Publications

Iron-specific Signal Separation from within Heavy Metal Stained Biological Samples Using X-Ray Microtomography with Polychromatic Source and Energy-Integrating Detectors.

Katchalski Tsvi T Case Tom T Kim Keun-Young KY Ramachandra Ranjan R Bushong Eric A EA Deerinck Thomas J TJ Haberl Matthias G MG Mackey Mason R MR Peltier Steven S Castillon Guillaume A GA Fujikawa Nobuko N Lawrence Albert R AR Ellisman Mark H MH

Scientific reports 20180515 1

Biological samples are frequently stained with heavy metals in preparation for examining the macro, micro and ultra-structure using X-ray microtomography and electron microscopy. A single X-ray microtomography scan reveals detailed 3D structure based on staining density, yet it lacks both material composition and functional information. Using a commercially available polychromatic X-ray source, energy integrating detectors and a two-scan configuration labelled by their energy- "High" and "Low", ...[more]

PMID: 29765060

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Project description:The objective of the study was to demonstrate that, in x-ray computed tomography (CT), more than two types of materials can be effectively separated with the use of an energy resolved photon-counting detector and classification methodology. Specifically, this applies to the case when contrast agents that contain K-absorption edges in the energy range of interest are present in the object. This separation is enabled via the use of recently developed energy resolved photon-counting detectors with multiple thresholds, which allow simultaneous measurements of the x-ray attenuation at multiple energies.To demonstrate this capability, we performed simulations and physical experiments using a six-threshold energy resolved photon-counting detector. We imaged mouse-sized cylindrical phantoms filled with several soft-tissue-like and bone-like materials and with iodine-based and gadolinium-based contrast agents. The linear attenuation coefficients were reconstructed for each material in each energy window and were visualized as scatter plots between pairs of energy windows. For comparison, a dual-kVp CT was also simulated using the same phantom materials. In this case, the linear attenuation coefficients at the lower kVp were plotted against those at the higher kVp.In both the simulations and the physical experiments, the contrast agents were easily separable from other soft-tissue-like and bone-like materials, thanks to the availability of the attenuation coefficient measurements at more than two energies provided by the energy resolved photon-counting detector. In the simulations, the amount of separation was observed to be proportional to the concentration of the contrast agents; however, this was not observed in the physical experiments due to limitations of the real detector system. We used the angle between pairs of attenuation coefficient vectors in either the 5-D space (for non-contrast-agent materials using energy resolved photon-counting acquisition) or a 2-D space (for contrast agents using energy resolved photon-counting acquisition and all materials using dual-kVp acquisition) as a measure of the degree of separation. Compared to dual-kVp techniques, an energy resolved detector provided a larger separation and the ability to separate different target materials using measurements acquired in different energy window pairs with a single x-ray exposure.We concluded that x-ray CT with an energy resolved photon-counting detector with more than two energy windows allows the separation of more than two types of materials, e.g., soft-tissue-like, bone-like, and one or more materials with K-edges in the energy range of interest. Separating material types using energy resolved photon-counting detectors has a number of advantages over dual-kVp CT in terms of the degree of separation and the number of materials that can be separated simultaneously.

Dataset Information

Iron-specific Signal Separation from within Heavy Metal Stained Biological Samples Using X-Ray Microtomography with Polychromatic Source and Energy-Integrating Detectors.

Publications

Iron-specific Signal Separation from within Heavy Metal Stained Biological Samples Using X-Ray Microtomography with Polychromatic Source and Energy-Integrating Detectors.

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