Electron probe x-ray microanalysis of subcellular ion transport in situ

1992 ◽  
Vol 50 (1) ◽  
pp. 16-17
Author(s):  
Avril V. Somlyo ◽  
Andrew P. Somlyo
Keyword(s):  
Electron Probe ◽  
High Electron ◽  
X Rays ◽  
Electron Dose ◽  
Single Spectrum ◽  
Thin Sections ◽  
Fast Electrons ◽  
X Ray

Electron probe x ray microanalysis [EPMA] provides quantitative information within a single spectrum about elements of biological interest with atomic number of 11 or greater. Therefore, the transport of ions and their accompanying co and counter ions across organelle membranes can be studied in situ by sampling within and adjacent to the intracellular organelle of interest under resting and stimulated conditions.EPMA is based on the fact that the ionization of atoms by fast electrons generates x rays having energies characteristic of the excited atoms. The interaction of incident fast electrons with atomic nuclei generates a background of continuum x rays. Elemental quantitation of ultra thin sections with EPMA is generally based on the linear relationship between elemental concentrations and the ratio of the number of characteristic/continuum. The use of this principle, together with the appropriate standards for calibration, has been the most successful approach for quantitative biological EPMA. The spatial resolution of EPMA at present is better than 10 nm and the practical limit of sensitivity for detecting calcium, (albeit with high electron dose), is approximately 0.3 mmol/kg dry wt. Two modes of data collection are utilized: fixed probe analysis of a region of interest or a scanning probe mode, where an x ray spectrum is collected at each picture point, to obtain quantitative elemental x ray maps. To preserve the morphology and the in vivo distribution of diffusible elements, we prepare specimens by rapid freezing in sub cooled Freon or, more recently with a Lifecell CF100 metal are mirror device; thin sections cut at -130 °C to -160 °C on a Reichert cryoultramicrotome. Msec time resolution of physiological are events can be achieved by freeze trapping.

scholarly journals Development of a sticker sealed microfluidic device for in situ analytical measurements using synchrotron radiation

2021 ◽  
Author(s):  
ITAMAR NECKEL ◽  
Lucas F. de Castro ◽  
Flavia Callefo ◽  
Verônica Teixeira ◽  
Angelo Gobbi ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
Green Energy ◽  
X Rays ◽  
Electrical Measurements ◽  
X Ray ◽  
Electrochemical Nucleation ◽  
Synchrotron Light

Abstract Shedding synchrotron light on microfluidic systems, exploring several contrasts in situ operando at the nanoscale, like X-ray fluorescence, diffraction, luminescence, and absorption, has the potential to reveal new properties and functionalities of materials across diverse areas, such as green energy, photonics, and nanomedicine. In this work, we present the micro-fabrication and characterization of a multifunctional polyester/glass sealed microfluidic device well-suited to combine with analytical X-ray techniques. The device consists of smooth microchannels patterned on glass, where three gold electrodes are deposited into the channels to serve in situ electrochemistry analysis or standard electrical measurements. It has been efficiently sealed through an ultraviolet-sensitive sticker-like layer based on a polyester film, and The burst pressure determined by pumping water through the microchannel(up to 0.22 MPa). Overall, the device has demonstrated exquisite chemical resistance to organic solvents, and its efficiency in the presence of biological samples (proteins) is remarkable. The device potentialities, and its high transparency to X-rays, have been demonstrated by taking advantage of the X-ray nanoprobe Carnaúba/Sirius/LNLS, by obtaining 2D X-ray nanofluorescence maps on the microchannel filled with water and after an electrochemical nucleation reaction. To wrap up, the microfluidic device characterized here has the potential to be employed in standard laboratory experiments as well as in situ and in vivo analytical experiments using a wide electromagnetic window, from infrared to X-rays, which could serve experiments in many branches of science.

scholarly journals Low angle x-ray diffraction studies of chromatin structure in vivo and in isolated nuclei and metaphase chromosomes

1983 ◽  
Vol 96 (4) ◽  
pp. 1120-1131 ◽  
Author(s):  
JP Langmore ◽  
Paulson JR
Keyword(s):  
Structural Integrity ◽  
Divalent Cations ◽  
Living Cells ◽  
X Rays ◽  
Metaphase Chromosomes ◽  
Thin Sections ◽  
Isolated Nuclei ◽  
X Ray

Diffraction of x-rays from living cells, isolated nuclei, and metaphase chromosomes gives rise to several major low angle reflections characteristic of a highly conserved pattern of nucleosome packing within the chromatin fibers. We answer three questions about the x-ray data: Which reflections are characteristic of chromosomes in vivo? How can these reflections be preserved in vitro? What chromosome structures give rise to the reflections? Our consistent observation of diffraction peaks at 11.0, 6.0, 3.8, 2.7 and 2.1 nm from a variety of living cells, isolated nuclei, and metaphase chromosomes establishes these periodicities as characteristic of eukaryotic chromosomes in vivo. In addition, a 30-40- nm peak is observed from all somatic cells that have substantial amounts of condensed chromatin, and a weak 18-nm reflection is observed from nucleated erythrocytes. These observations provide a standard for judging the structural integrity of isolated nuclei, chromosomes, and chromatin, and thus resolve long standing controversy about the "tru" nature of chromosome diffraction. All of the reflection seen in vivo can be preserved in vitro provided that the proper ionic conditions are maintained. Our results show clearly that the 30-40-nm maximum is a packing reflection. The packing we observe in vivo is directly correlated to the side-by-side arrangement of 20- 30-nm fibers observed in thin sections of fixed and dehydrated cells and isolated chromosomes. This confirms that such packing is present in living cells and is not merely an artifact of electron microscopy. As expected, the packing reflection is shifted to longer spacings when the fibers are spread apart by reducing the concentration of divalent cations in vitro. Because the 18-, 11.0-, 6.0-, 3.8-, 2.7-, and 2.1-nm reflections are not affected by the decondensation caused by removal of divalent cations, these periodicities must reflect the internal structure of the chromaticn fibers.

scholarly journals Development of a sticker sealed microfluidic device for in situ analytical measurements using synchrotron radiation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Itamar T. Neckel ◽  
Lucas F. de Castro ◽  
Flavia Callefo ◽  
Verônica C. Teixeira ◽  
Angelo L. Gobbi ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
Green Energy ◽  
X Rays ◽  
Electrical Measurements ◽  
X Ray ◽  
Electrochemical Nucleation ◽  
Synchrotron Light

AbstractShedding synchrotron light on microfluidic systems, exploring several contrasts in situ/operando at the nanoscale, like X-ray fluorescence, diffraction, luminescence, and absorption, has the potential to reveal new properties and functionalities of materials across diverse areas, such as green energy, photonics, and nanomedicine. In this work, we present the micro-fabrication and characterization of a multifunctional polyester/glass sealed microfluidic device well-suited to combine with analytical X-ray techniques. The device consists of smooth microchannels patterned on glass, where three gold electrodes are deposited into the channels to serve in situ electrochemistry analysis or standard electrical measurements. It has been efficiently sealed through an ultraviolet-sensitive sticker-like layer based on a polyester film, and The burst pressure determined by pumping water through the microchannel(up to 0.22 MPa). Overall, the device has demonstrated exquisite chemical resistance to organic solvents, and its efficiency in the presence of biological samples (proteins) is remarkable. The device potentialities, and its high transparency to X-rays, have been demonstrated by taking advantage of the X-ray nanoprobe Carnaúba/Sirius/LNLS, by obtaining 2D X-ray nanofluorescence maps on the microchannel filled with water and after an electrochemical nucleation reaction. To wrap up, the microfluidic device characterized here has the potential to be employed in standard laboratory experiments as well as in in situ and in vivo analytical experiments using a wide electromagnetic window, from infrared to X-rays, which could serve experiments in many branches of science.

Application of the Electron Probe Microanalyzer to the Analysis of Planetary and Geological Materials: a Historical Perspective.

1999 ◽  
Vol 5 (S2) ◽  
pp. 572-573 ◽  
Author(s):  
Klaus Keil
Keyword(s):  
Electron Probe ◽  
Grain Morphology ◽  
X Rays ◽  
Thin Sections ◽  
Electron Probe Microanalyzer ◽  
Excitation Volume ◽  
Quantitative Analyses ◽  
Short Time ◽  
Non Destructive

Castaing first presented his idea to use secondary X-rays excited by a focused electron beam from a polished solid sample for microanalysis in 1949 at the First European Regional Conference on Electron Microscopy in Delft, Netherlands. As part of his dissertation, he then not only built the first electron probe microanalyzer (EPM), but also established many of the theoretical and analytical principles of the technique. This technique offered enormous analytical advantages to earth scientists over other analytical methods available at the time. For example, it allows qualitative and quantitative analysis of individual mineral grains a few microns in diameter; mineral grains can be viewed during analysis, thus ensuring accurate correlation between composition and grain morphology; for most purposes, the method is non-destructive; in situ analysis of minerals in polished thin sections results in retention of textural relationships among coexisting minerals; because of the small excitation volume, the technique is ideally suited for the study of zoned minerals, minute inclusions, exsolution lamellae, etc.; and once suitable standards are prepared and correction procedures are established, a large number of quantitative analyses can be obtained in a comparatively short time.

High spatial resolution x-ray microanalysis of interfaces

1995 ◽  
Vol 53 ◽  
pp. 284-285
Author(s):  
J. R. Michael
Keyword(s):  
Spatial Resolution ◽  
Electron Probe ◽  
Solid Angle ◽  
Collection Efficiency ◽  
Spatial Scales ◽  
Energy Dispersive Spectrometer ◽  
X Rays ◽  
X Ray ◽  
Probe Size ◽  
Brightness Field

X-ray microanalysis in the analytical electron microscope (AEM) refers to a technique by which chemical composition can be determined on spatial scales of less than 10 nm. There are many factors that influence the quality of x-ray microanalysis. The minimum probe size with sufficient current for microanalysis that can be generated determines the ultimate spatial resolution of each individual microanalysis. However, it is also necessary to collect efficiently the x-rays generated. Modern high brightness field emission gun equipped AEMs can now generate probes that are less than 1 nm in diameter with high probe currents. Improving the x-ray collection solid angle of the solid state energy dispersive spectrometer (EDS) results in more efficient collection of x-ray generated by the interaction of the electron probe with the specimen, thus reducing the minimum detectability limit. The combination of decreased interaction volume due to smaller electron probe size and the increased collection efficiency due to larger solid angle of x-ray collection should enhance our ability to study interfacial segregation.

scholarly journals Optimization of X-ray Investigations in Dentistry Using Optical Coherence Tomography

Sensors ◽  
2021 ◽  
Vol 21 (13) ◽  
pp. 4554
Author(s):  
Ralph-Alexandru Erdelyi ◽  
Virgil-Florin Duma ◽  
Cosmin Sinescu ◽  
George Mihai Dobre ◽  
Adrian Bradu ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Radiation Dose ◽  
Imaging Techniques ◽  
Image Resolution ◽  
Optical Coherence ◽  
X Rays ◽  
High Quality ◽  
X Ray

The most common imaging technique for dental diagnoses and treatment monitoring is X-ray imaging, which evolved from the first intraoral radiographs to high-quality three-dimensional (3D) Cone Beam Computed Tomography (CBCT). Other imaging techniques have shown potential, such as Optical Coherence Tomography (OCT). We have recently reported on the boundaries of these two types of techniques, regarding. the dental fields where each one is more appropriate or where they should be both used. The aim of the present study is to explore the unique capabilities of the OCT technique to optimize X-ray units imaging (i.e., in terms of image resolution, radiation dose, or contrast). Two types of commercially available and widely used X-ray units are considered. To adjust their parameters, a protocol is developed to employ OCT images of dental conditions that are documented on high (i.e., less than 10 μm) resolution OCT images (both B-scans/cross sections and 3D reconstructions) but are hardly identified on the 200 to 75 μm resolution panoramic or CBCT radiographs. The optimized calibration of the X-ray unit includes choosing appropriate values for the anode voltage and current intensity of the X-ray tube, as well as the patient’s positioning, in order to reach the highest possible X-rays resolution at a radiation dose that is safe for the patient. The optimization protocol is developed in vitro on OCT images of extracted teeth and is further applied in vivo for each type of dental investigation. Optimized radiographic results are compared with un-optimized previously performed radiographs. Also, we show that OCT can permit a rigorous comparison between two (types of) X-ray units. In conclusion, high-quality dental images are possible using low radiation doses if an optimized protocol, developed using OCT, is applied for each type of dental investigation. Also, there are situations when the X-ray technology has drawbacks for dental diagnosis or treatment assessment. In such situations, OCT proves capable to provide qualitative images.

In Situ Testing and X Rays Radiation Effects of Pt/Bi3.15Nd0.85Ti3O12/Pt Ferroelectric Capacitors

2013 ◽  
Vol 712-715 ◽  
pp. 293-297
Author(s):  
Li Li
Keyword(s):  
Radiation Effects ◽  
Hysteresis Loops ◽  
Electrical Performance ◽  
Hysteresis Curve ◽  
X Rays ◽  
Shanghai Synchrotron Radiation Facility ◽  
X Ray ◽  
In The Beginning ◽  
Ferroelectric Capacitors

Pt/Bi3.15Nd0.85Ti3O12(BNT)/Pt ferroelectric capacitors were monitored using in situ X-ray irradiation with 10 keV at BL14B1 beamline (Shanghai Synchrotron Radiation Facility). BL14B1 combined with a ferroelectric analyzer enabled measurements in situ of electrical performance. The hysteresis curve (PE) of distortion depended on the polarization during irradiation, but the diffracted intensities of the (117) peak did not change in the beginning. ThePEcurve had a negligible change from 2.09×109Gy to 4.45×109Gy. Finally, bothPrandPr+very rapidly increased, but the intensities of (117) decreased. The hysteresis loops were remarkably deformed at the maximum total dose of 4.87×109Gy.

scholarly journals THE EFFECTS OF ROENTGEN RAYS ON CELL-VIRUS ASSOCIATIONS

1943 ◽  
Vol 78 (4) ◽  
pp. 285-304 ◽  
Author(s):  
William F. Friedewald ◽  
Rubert S. Anderson
Keyword(s):  
X Rays ◽  
Pathological Changes ◽  
Cellular Division ◽  
Papilloma Virus ◽  
X Ray ◽  
Crude Extracts ◽  
Domestic Rabbits ◽  
Roentgen Rays

The virus-induced papillomas of cottontail as well as domestic rabbits regress completely within a few weeks when exposed to 5,000 r of x-ray irradiation. The x-rays do not immediately kill the papilloma cells, but lead to death by inhibiting cellular division and producing pathological changes in the cells which then continue to differentiate. The virus associated with the growths, however, not only persists in undiminished amount during regression, but often an increased yield of it can be obtained on extraction. The fibroma virus in crude extracts or in vivo is inactivated by far less irradiation than the papilloma virus. 10,000 r destroys 90 per cent or more of the infectivity of the fibroma virus, whereas at least 100,000 r is required to inactivate 50 per cent of the papilloma virus in extracts containing about the same amount of protein. No variant of the papilloma virus or fibroma virus has been encountered as a result of the irradiation.

Applications of High-Resolution Powder X-Ray Diffraction

2007 ◽  
Vol 130 ◽  
pp. 7-14 ◽  
Author(s):  
Andrew N. Fitch
Keyword(s):  
High Resolution ◽  
Synchrotron Radiation ◽  
Powder Diffraction ◽  
X Rays ◽  
X Ray Diffraction ◽  
Crystalline Materials ◽  
X Ray ◽  
Structural Characterisation ◽  
Pdf Analysis

The highly-collimated, intense X-rays produced by a synchrotron radiation source can be harnessed to build high-resolution powder diffraction instruments with a wide variety of applications. The general advantages of using synchrotron radiation for powder diffraction are discussed and illustrated with reference to the structural characterisation of crystalline materials, atomic PDF analysis, in-situ and high-throughput studies where the structure is evolving between successive scans, and the measurement of residual strain in engineering components.

scholarly journals In situ damage assessment using synchrotron X-rays in materials loaded by a Hopkinson bar

2014 ◽  
Vol 372 (2015) ◽  
pp. 20130191 ◽  
Author(s):  
Weinong W. Chen ◽  
Matthew C. Hudspeth ◽  
Ben Claus ◽  
Niranjan D. Parab ◽  
John T. Black ◽  
...  
Keyword(s):  
High Speed ◽  
Particle Interaction ◽  
Tensile Failure ◽  
High Rate ◽  
X Rays ◽  
Contrast Imaging ◽  
X Ray ◽  
Damage Initiation ◽  
Dynamic Experiments

Split Hopkinson or Kolsky bars are common high-rate characterization tools for dynamic mechanical behaviour of materials. Stress–strain responses averaged over specimen volume are obtained as a function of strain rate. Specimen deformation histories can be monitored by high-speed imaging on the surface. It has not been possible to track the damage initiation and evolution during the dynamic deformation inside specimens except for a few transparent materials. In this study, we integrated Hopkinson compression/tension bars with high-speed X-ray imaging capabilities. The damage history in a dynamically deforming specimen was monitored in situ using synchrotron radiation via X-ray phase contrast imaging. The effectiveness of the novel union between these two powerful techniques, which opens a new angle for data acquisition in dynamic experiments, is demonstrated by a series of dynamic experiments on a variety of material systems, including particle interaction in granular materials, glass impact cracking, single crystal silicon tensile failure and ligament–bone junction damage.

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