Imaging Plate Scanner Research Articles

Absolute x-ray calibration of an Amersham imaging plate scanner.

The absolute response of the GE Amersham Typhoontm imaging plate scanner is studied in this paper. The sensitivity function of the scanner with different photomultiplier tube voltages was obtained by using a pre-calibrated Cu Kα x-ray tube. The results showed that the sensitivity function decreases exponentially with higher voltage and is also affected by the scanning pixel size. The spatial resolution and the fading effect of the imaging plate system on x rays were also investigated and compared with the previous scanner models.

Structure factors and charge-density study of diamond at 800 K

The structure factors of diamond were determined by synchrotron radiation X-ray powder diffraction at 800 K at sin θ/λ ≤ 2.2 Å−1 reciprocal resolution. The structure factors were estimated using six powder profiles measured on beamline BL02B2 at SPring-8 (Hyogo, Japan). A high reciprocal resolution at sin θ/λ ≤ 2.2 Å−1 was required to reveal the temperature dependence of the charge density, due to the high Debye temperature of θD = 1860 K of diamond. Wide 2θ angle data with the highest counting statistics are crucial for accurate data analysis. The periodic noise of every six-pixel step was observed in the highest counting statistics imaging plate (IP) data scanned by a BAS2500 IP scanner. It was found that the noise originated from the six-sided polygonal mirror in the scanner. The intensity fluctuation at every six-pixel step was also found in the Fourier series expansion of the powder profiles. The ratio of the maximum fluctuation was estimated as 0.4% by summing all six-pixel step data. The powder profiles were corrected by multiplying the ratios. The intensity fluctuation in the background region was reduced to less than 50% of the uncorrected data. The weak 888 Bragg reflection, with an intensity of 0.005% of that of the 111 Bragg reflection at 800 K, was readily observed in the corrected data. Finally, the structure factors determined at 800 K were successfully applied to a charge-density study by multipole modelling. The reliability factors and multipole parameters at 800 K are in agreement with those at 300 K. The differences in the charge density at the bond midpoint and ∇2ρ at the bond-critical point were less than 1% and 2%, respectively.

Cross-calibration of Fuji TR image plate and RAR 2492 x-ray film to determine the response of a DITABIS Super Micron image plate scanner.

Fuji TR image plate is frequently used as a replacement detector medium for x-ray imaging and spectroscopy diagnostics at NIF, Omega, and Z facilities. However, the familiar Fuji BAS line of image plate scanners is no longer supported by the industry, and so a replacement scanning system is needed. While the General Electric Typhoon line of scanners could replace the Fuji systems, the shift away from photo stimulated luminescence units to 16-bit grayscale Tag Image File Format (TIFF) leaves a discontinuity when comparing data collected from both systems. For the purposes of quantitative spectroscopy, a known unit of intensity applied to the grayscale values of the TIFF is needed. The DITABIS Super Micron image plate scanning system was tested and shown to potentially rival the resolution and dynamic range of Kodak RAR 2492 x-ray film. However, the absolute sensitivity of the scanner is unknown. In this work, a methodology to cross calibrate Fuji TR image plate and the absolutely calibrated Kodak RAR 2492 x-ray film is presented. Details of the experimental configurations used are included. An energy dependent scale factor to convert Fuji TR IP scanned on a DITABIS Super Micron scanner from 16-bit grayscale TIFF to intensity units (i.e., photons per square micron) is discussed.

X射线成像板衰退曲线标定

Based on the X-ray diffractometer in the lab,the calibration experiment of the fading curve of the imaging plate(IP)was introduced.The type of the sample was Fuji SR-2025and the concerned signals were X-rays.A Cu X-ray tube was used as the X-ray source in the experiment while a trihydroxymethylaminomethane(TAM)crystal was used to provide Kαmonochromatic radiations.The type of IP scanner was BAS-5000and the environment temperature was(20±1)℃.Under the conditions that the X-rays were stable,the spatial uniformity and the line response of the IP were examined well,and the fading curve of the IP was obtained by exposing the IP on different place in different time.Fitting analysis of the fading curve was carried out and the uncertainty was discussed.The experimental results show that the instability of X-ray source is 0.7%and the spatial uniformity of IP is less than 1%;excellent linear response of the sample is also proved;at the energy point of Cu-Kα8 027.84eV,the fading curve of the IP decreases slowly in exponential formη(t)=0.36884·exp(-t/159.647 56)+0.633 72;the recorded signals remain 80% while 125minutes after exposure with visible light shielding.

X射线成像板衰退曲线标定

Based on the X-ray diffractometer in the lab,the calibration experiment of the fading curve of the imaging plate(IP)was introduced.The type of the sample was Fuji SR-2025and the concerned signals were X-rays.A Cu X-ray tube was used as the X-ray source in the experiment while a trihydroxymethylaminomethane(TAM)crystal was used to provide Kαmonochromatic radiations.The type of IP scanner was BAS-5000and the environment temperature was(20±1)℃.Under the conditions that the X-rays were stable,the spatial uniformity and the line response of the IP were examined well,and the fading curve of the IP was obtained by exposing the IP on different place in different time.Fitting analysis of the fading curve was carried out and the uncertainty was discussed.The experimental results show that the instability of X-ray source is 0.7%and the spatial uniformity of IP is less than 1%;excellent linear response of the sample is also proved;at the energy point of Cu-Kα8 027.84eV,the fading curve of the IP decreases slowly in exponential formη(t)=0.36884·exp(-t/159.647 56)+0.633 72;the recorded signals remain 80% while 125minutes after exposure with visible light shielding.

High-energy X-ray diffraction using the Pixium 4700 flat-panel detector

The Pixium 4700 detector represents a significant step forward in detector technology for high-energy X-ray diffraction. The detector design is based on digital flat-panel technology, combining an amorphous Si panel with a CsI scintillator. The detector has a useful pixel array of 1910 x 2480 pixels with a pixel size of 154 microm x 154 microm, and thus it covers an effective area of 294 mm x 379 mm. Designed for medical imaging, the detector has good efficiency at high X-ray energies. Furthermore, it is capable of acquiring sequences of images at 7.5 frames per second in full image mode, and up to 60 frames per second in binned region of interest modes. Here, the basic properties of this detector applied to high-energy X-ray diffraction are presented. Quantitative comparisons with a widespread high-energy detector, the MAR345 image plate scanner, are shown. Other properties of the Pixium 4700 detector, including a narrow point-spread function and distortion-free image, allows for the acquisition of high-quality diffraction data at high X-ray energies. In addition, high frame rates and shutterless operation open new experimental possibilities. Also provided are the necessary data for the correction of images collected using the Pixium 4700 for diffraction purposes.

X-ray modulation transfer functions of photostimulable phosphor image plates and scanners

The modulation transfer functions of two types of photostimulable phosphor image plates were determined in the 10 keV to 50 keV x-ray energy range using a resolution test pattern with up to 10 line pairs per mm (LP/mm) and a wavelength dispersive x-ray spectrometer. Techniques were developed for correcting for the partial transmittance of the high energy x rays through the lead bars of the resolution test pattern, and the modulation transfer function (MTF) was determined from the measured change in contrast with LP/mm values. The MTF was convolved with the slit function of the image plate scanner, and the resulting point spread functions (PSFs) were in good agreement with the observed shapes and widths of x-ray spectral lines and with the PSF derived from edge spread functions. The shapes and the full width at half-maximum (FWHM) values of the PSF curves of the Fuji Superior Resolution (SR) and Fuji Maximum Sensitivity (MS) image plate detectors, consisting of the image plate and the scanner, determined by the three methods gave consistent results: The SR PSF is Gaussian with 0.13 mm FWHM, and the MS PSF is Lorentzian with 0.19 mm FWHM. These techniques result in the accurate determination of the spatial resolution achievable using image plate and scanner combinations and enable the optimization of spatial resolution for x-ray spectroscopy and radiography.

Optimisation of lithium borate–barium chloride glass-ceramic thermal neutron imaging plates

Glass-ceramic thermal neutron imaging plates (NIPs) recently reported have been further developed for use in neutron radiography. The plate consists of nanocrystallites of the storage phosphor BaCl 2:Eu 2+ embedded within a neutron-sensitive lithium borate glass-matrix. A new generation of samples, enriched with both 10B and 6Li with thicknesses in the range 280–500 μm have been studied. Neutron images were read out using a Fuji BAS2500 imaging plate scanner and the quality of the images obtained was comparable to those recorded on a commercial NIP. Details of the response to neutron-irradiation as well as the obtained spatial resolution of the images are presented. The neutron absorption and radiation hardness of the materials studied is also measured.

The small-angle and wide-angle X-ray scattering set-up at beamline BL9 of DELTA

The multi-purpose experimental endstation of beamline BL9 at the Dortmund Electron Accelerator (DELTA) is dedicated to diffraction experiments in grazing-incidence geometry, reflectivity and powder diffraction measurements. Moreover, fluorescence analysis and inelastic X-ray scattering experiments can be performed. Recently, a new set-up for small-angle and wide-angle X-ray scattering utilizing detection by means of an image-plate scanner was installed and is described in detail here. First small-angle X-ray scattering experiments on aqueous solutions of lysozyme with different cosolvents and of staphylococcal nuclease are discussed. The application of the set-up for texture analysis is emphasized and a study of the crystallographic texture of natural bio-nanocomposites, using lobster and crab cuticles as model materials, is presented.

Structure and neutron imaging characteristics of lithium borate–barium chloride glass-ceramics

A glass-ceramic has been developed which can be used as a thermal neutron imaging plate (NIP) for neutron radiography. The plate consists of nanocrystallites of the storage phosphor BaCl 2:Eu 2+ embedded within a neutron sensitive lithium borate glass-matrix. Details of their preparation and structure are discussed. Neutron images were read out using a Fuji BAS2500 imaging plate scanner and the quality of the images obtained is comparable to those recorded on a commercial NIP. Details of the response to neutron and X-irradiation as well as the obtained spatial resolution of the images are presented. The neutron activation of the materials studied is also calculated.

A Novel Arabidopsis Gene Causes Bax-like Lethality in Saccharomyces cerevisiae

Overexpression of the mammalian proapoptotic protein Bax induces cell death in plant and yeast cells. The Bax inihibitor-1 (BI-1) gene rescues yeast and plant from Bax-mediated lethality. Using the Arabidopsis BI-1 (AtBI-1) gene controlled by the GAL1 promoter as a cell death suppressor in yeast, Cdf1 (cell growth defect factor-1) was isolated from Arabidopsis cDNA library. Overexpression of Cdf1 caused cell death in yeast, whereas such an effect was suppressed by co-expression of AtBI-1. The Cdf1 protein fused with a green fluorescent protein was localized in the mitochondria and resulted in the loss of mitochondrial membrane potential in yeast. The Bax-resistant mutant BRM1 demonstrated tolerance against Cdf1-mediated lethality, whereas the Deltaatp4 strain was sensitive to Cdf1. Our results suggest that Cdf1 and Bax cause mitochondria-mediated yeast lethality through partially overlapped pathways.

Structure of the zinc-saturated C-terminal lobe of bovine lactoferrin at 2.0 Å resolution

The crystal structure of the zinc-saturated C-terminal lobe of bovine lactoferrin has been determined at 2.0 A resolution using crystals stabilized at pH 3.8. This is the first metal-saturated structure of any functional lactoferrin at such a low pH. Purified samples of proteolytically generated zinc-saturated C-terminal lobe were crystallized from 0.1 M MES buffer pH 6.5 containing 25%(v/v) polyethyleneglycol monomethyl ether 550 and 0.1 M zinc sulfate heptahydrate. The crystals were transferred to 25 mM ammonium acetate buffer containing 25%(v/v) polyethyleneglycol monomethyl ether 550 and the pH was gradually changed from 6.5 to 3.8. The X-ray intensity data were collected with a 345 mm imaging-plate scanner mounted on an RU-300 rotating-anode X-ray generator using crystals soaked in the buffer at pH 3.8. The structure was determined with the molecular-replacement method using the coordinates of the monoferric C-terminal lobe of bovine lactoferrin as a search model and was refined to an R factor of 0.192 for all data to 2.0 A resolution. The final model comprises 2593 protein atoms (residues 342-676 and 681-685), 138 carbohydrate atoms (from 11 monosaccharide units in three glycan chains), three Zn2+ ions, one CO3(2-) ion, one SO(4)2- ions and 227 water molecules. The overall folding of the present structure is essentially similar to that of the monoferric C-terminal lobe of bovine lactoferrin, although it contains Zn2+ in place of Fe3+ in the metal-binding cleft as well as two additional Zn2+ ions on the surface of the C-terminal lobe. The Zn2+ ion in the cleft remains bound to the lobe with octahedral coordination. The bidentate carbonate ion is stabilized by a network of hydrogen bonds to Ala465, Gly466, Thr459 and Arg463. The other two zinc ions also form sixfold coordinations involving symmetry-related protein and water molecules. The number of monosaccharide residues from the three glycan chains of the C-terminal lobe was 11, which is the largest number observed to date. The structure shows that the C-terminal lobe of lactoferrin is capable of sequestering a Zn2+ ion at a pH of 3.8. This implies that the zinc ions can be sequestered over a wide pH range. The glycan chain attached to Asn545 may also have some influence on iron release from the C-terminal lobe.

A new fast and large area neutron detector using a novel image plate readout technique

At Agfa Medical Imaging Systems, a new type of image plate scanner is being developed. Instead of scanning the irradiated image plate with a single laser and reading the luminescence information with a single light collector and photomultiplier, the new system employs a whole line of laser diodes and a set of CCD line sensors. This technique allows for virtually unlimited detection areas and a very fast readout.

Crystal Structure of a Proteolytically Generated Functional Monoferric C-lobe of Bovine Lactoferrin at 1.9 Å Resolution

This is the first crystal structure of a proteolytically generated functional C-lobe of lactoferrin. The purified samples of iron-saturated C-lobe were crystallized in 0.1 M Mes buffer (pH 6.5) containing 25% (v/v) polyethyleneglycol monomethyl ether 550 M and 0.1 M zinc sulphate heptahydrate. The X-ray intensity data were collected with 300 mm imaging plate scanner mounted on a rotating anode generator. The structure was determined by the molecular replacement method using the coordinates of the C-terminal half of bovine lactoferrin as a search model and refined to an R-factor of 0.193 for all data to 1.9 Å resolution. The final model comprises 2593 protein atoms (residues 342–676 and 681–685), 124 carbohydrate atoms (from ten monosaccharide units, in three glycan chains), one Fe 3+, one CO 3 2−, two Zn 2+ and 230 water molecules. The overall folding of the C-lobe is essentially the same as that of C-terminal half of bovine lactoferrin but differs slightly in conformations of some of the loops and reveals a number of new interactions. There are 20 Cys residues in the C-lobe forming ten disulphide links. Out of these, one involving Cys481-Cys675 provides an inter-domain link at 2.01 Å while another Cys405-Cys684 is formed between the main C-lobe 342-676 and the hydrolyzed pentapeptide 681-685 fragment. Six inter-domain hydrogen bonds have been observed in the structure whereas only four were reported in the structure of intact lactoferrin, although domain orientations have been found similar in the two structures. The good quality of electron density has also revealed all the ten oligosaccharide units in the structure. The observation of two metal ions at sites other than the iron-binding cleft is another novel feature of the present structure. These zinc ions stabilize the crystal packing. This structure is also notable for extensive inter-molecular hydrogen bonding in the crystals. Therefore, the present structure appears to be one of the best packed crystal structures among the proteins of the transferrin superfamily.

High-pressure protein crystallography (HPPX): instrumentation, methodology and results on lysozyme crystals.

A new set-up and associated methodology for the collection of angle-dispersive diffraction data from protein crystals submitted to high hydrostastic pressure have been developed on beamline ID30 at the ESRF. The instrument makes use of intense X-rays of ultra-short wavelength emitted by two collinear undulators, and combines a membrane-driven diamond-anvil cell mounted on a two-axis goniometer and an imaging-plate scanner. Sharp and clean diffraction pictures from tetragonal crystals of hen egg-white lysozyme (tHEWL) and orthorhombic crystals of bovine erythrocyte Cu, Zn superoxide dismutase (SOD) were recorded at room temperature and pressures up to 0.915 and 1.00 GPa, respectively. The compressibility of tHEWL was determined from unit-cell parameters determined at 24 different pressures up to 0.915 GPa. High-pressure diffraction data sets from several crystals of tHEWL were collected and analyzed. Merging of data recorded on different crystals at 0.30 and 0.58 GPa produced two sets of structure amplitudes with good resolution, completeness, redundancy and R(sym) values. A third set at 0.69 GPa was of a similar quality except a lower completeness. The three structures have been refined. The pressure-induced loss of crystalline order in a tHEWL crystal beyond 0.82 GPa was captured through a series of diffraction pictures.

Overview of the tunable beamlines for protein crystallography at the EMBL Hamburg Outstation; an analysis of current and future usage and developments

The EMBL Hamburg Outstation currently operates two tunable protein crystallography beamlines suitable for single and multiple anomalous diffraction (SAD/MAD) experiments. The first beamline, designated X31, is located on a bending magnet of the DORIS III storage ring whereas the second beamline, BW7A, is positioned at a multipole wiggler at the same storage ring. X31 is equipped with an energy stabilization device to ensure constant wavelength during longer data-collection periods. The in-house built crystallographic end-station is now equipped with a Mar345 imaging-plate scanner as a detector. The wiggler beamline BW7A features a novel sagitally focusing monochromator. The end-station used here has also been developed and built in-house. The beamline is currently operated with a Mar 165 CCD detector. In this paper the hardware and software developments of the last years will be summarized and the outlook for substantial upgrades will be given. The future plans include the design and construction of a third tunable beamline, designated X12, for protein crystallography. The development of automated beamlines for protein crystallography is of particular importance with respect to structural genomics initiatives. The analysis of the projects of the last years shows the wide range of anomalous scatterer used on the tunable beamlines thus demonstrating the need of a wide range of accessible energies and fast and reliable energy changes.

Fiber diffraction using the BioCAT undulator beamline at the Advanced Photon Source

The BioCAT undulator-based beamline at the Advanced Photon Source, Argonne IL, USA is designed to be a state-of-the-art instrument for biological non-crystalline diffraction and X-ray absorption spectroscopy. The optics consist of double crystal monochromators with sagitally focussing second crystals followed by a vertically focussing mirror which allow independent focussing of the beam in the vertical and horizontal directions virtually anywhere along the length of the 12 m experimental enclosure. When configured for a 2 m fiber diffraction camera, a focal spot of less than 40×200 μm (FWHM) has been observed which contained essentially all of the 1.5–2.5×10 13 ph/s delivered by the cryogenically-cooled Si(1 1 1) double crystal monochromator. This combination of highly demagnifying optics and the very low divergence of the very small source have yielded excellent quality patterns from various muscle specimens and collagen-containing tissues. Detectors available include a Fuji BAS2500 image plate scanner and a 1 k×1 k CCD detector optimized for small-angle applications. Future developments will include, in vacuum beam monitoring, longer camera lengths (6–8 m), and optimizations to improve first-order resolution in small-angle applications.

New high-DQE imaging plate scanner using the reflected readout laser signal for noise corrections.

Imaging Plates (IPs) are in principle ideal electron detectors combining a large active layer area with a high sensitivity, linear dynamic range detection over 5 orders of magnitude. A moderate resolution and a decreasing detection quantum efficiency (DQE) for higher electron doses limit their use so far. The decrease of the DQE results from linear noise contributed by readout laser instabilities and inhomogeneities of the IP active layer. Here we present data on a new IP drum scanner prototype. This scanner combines twin channel amplification electronics with a new type of readout laser which allows a smaller readout focus and increased stability. The current nominal pixel size is 25 microm, and the measured modulation transfer function (MTF) indicates that further reduction of the scanning step size down to pixel sizes in the range of 12-15 microm should be possible. A unique feature of the new scanner is the simultaneous recording of the reflected readout laser light. The reflected light signal can be used for a posteriori alignment of repeated scans of one individual IP and for a correction of one part of the high spatial frequency noise contribution (reflected light correction). The posteriori alignment now allows an easy conventional gain normalization of the luminescence signal without using special markers on the IP. Both corrections lead to an increase of the DQE for high electron doses.

Refinement and structural analysis of barnase at 1.5 A resolution.

The structure of Bacillus amyloliquefaciens ribonuclease (barnase), an extracellular 110-residue enzyme initially solved at 2.0 A resolution, has been refined at 1.5 A using synchrotron radiation and an imaging-plate scanner. Refinement with anisotropic atomic displacement parameters resulted in increased accuracy of the structure. The final model has a crystallographic R factor of 11.5% and an Rfree of 17.4%. The three independent molecules in the asymmetric unit, referred to as A, B and C, allowed detailed analysis of this final model and meaningful comparison with structures of barnase complexed either with nucleotide inhibitors or with its natural intracellular inhibitor, barstar. The analysis of the overall solvent structure revealed a similar number of water molecules associated with each barnase molecule; among these were 16 equivalent buried solvent molecules, the locations of which are discussed in detail and classified on the basis of their structural role. The importance of the water molecules' contribution to the barnase-barstar interaction is also highlighted. The high accuracy of the present analysis revealed the presence of a Zn2+ ion mediating the contacts between pairs of symmetry-related A, B or C molecules; such an ion had previously only been identified for pairs of C molecules.

Crystal growth and preliminary X-ray study of glutamic acid specific serine protease from Bacillus intermedius

The glutamic acid specific protease (glutamyl-endopeptidase) from Bacillus intermedius, strain 3-19, was isolated and purified using ion exchange chromatography on CM-cellulose and Mono-S FPLC column. The conditions for crystallization of the enzyme have been discussed. The crystals of enzyme were grown using hanging-drop vapor-diffusion technique. Crystals belong to the space group C2 with unit cell parameters of a=61.62 Å, b=55.84 Å, c=60.40 Å, β=117.6° X-ray diffraction data to 1.68 Å resolution were collected using synchrotron radiation (EMBL, Hamburg) and an imaging plate scanner.