Project description:We report the determination of the structure of Escherichia coli ?-galactosidase at a resolution of ?1.8?Å using data collected on a 200?kV CRYO ARM microscope equipped with a K3 direct electron detector. The data were collected in a single 24?h session by recording images from an array of 7 × 7 holes at each stage position using the automated data collection program SerialEM. In addition to the expected features such as holes in the densities of aromatic residues, the map also shows density bumps corresponding to the locations of hydrogen atoms. The hydrogen densities are useful in assigning absolute orientations for residues such as glutamine or asparagine by removing the uncertainty in the fitting of the amide groups, and are likely to be especially relevant in the context of structure-guided drug design. These findings validate the use of electron microscopes operating at 200?kV for imaging protein complexes at atomic resolution using cryo-EM.
Project description:To measure and investigate the improvement of microcalcification (MC) visibility in cone beam breast CT with a high pitch (75 μm), thick (500 μm) scintillator CMOS/CsI flat panel detector (Dexela 2923, Perkin Elmer).Aluminum wires and calcium carbonate grains of various sizes were embedded in a paraffin cylinder to simulate imaging of calcifications in a breast. Phantoms were imaged with a benchtop experimental cone beam CT system at various exposure levels. In addition to the Dexela detector, a high pitch (50 μm), thin (150 μm) scintillator CMOS/CsI flat panel detector (C7921CA-09, Hamamatsu Corporation, Hamamatsu City, Japan) and a widely used low pitch (194 μm), thick (600 μm) scintillator aSi/CsI flat panel detector (PaxScan 4030CB, Varian Medical Systems) were also used in scanning for comparison. The images were independently reviewed by six readers (imaging physicists). The MC visibility was quantified as the fraction of visible MCs and measured as a function of the estimated mean glandular dose (MGD) level for various MC sizes and detectors. The modulation transfer functions (MTFs) and detective quantum efficiencies (DQEs) were also measured and compared for the three detectors used.The authors have demonstrated that the use of a high pitch (75 μm) CMOS detector coupled with a thick (500 μm) CsI scintillator helped make the smaller 150-160, 160-180, and 180-200 μm MC groups more visible at MGDs up to 10.8, 9, and 10.8 mGy, respectively. It also made the larger 200-212 and 212-224 μm MC groups more visible at MGDs up to 7.2 mGy. No performance improvement was observed for 224-250 μm or larger size groups. With the higher spatial resolution of the Dexela detector based system, the apparent dimensions and shapes of MCs were more accurately rendered. The results show that with the aforementioned detector, a 73% visibility could be achieved in imaging 160-180 μm MCs as compared to 28% visibility achieved by the low pitch (194 μm) aSi/CsI flat panel detector. The measurements confirm that the Hamamatsu detector has the highest MTF, followed by the Dexel detector, and then the Varian detector. However, the Dexela detector, with its thick (500 μm) CsI scintillator and low noise level, has the highest DQE at all frequencies, followed by the Varian detector, and then the Hamamatsu detector. The findings on the MC visibility correlated well with the differences in MTFs, noise power spectra, and DQEs measured for these three detectors.The authors have demonstrated that the use of the CMOS type Dexela detector with its high pitch (75 μm) and thick (500 μm) CsI scintillator could help improve the MC visibility. However, the improvement depended on the exposure level and the MC size. For imaging larger MCs or scanning at high exposure levels, there was little advantage in using the Dexela detector as compared to the aSi type Varian detector. These findings correlate well with the higher measured DQEs of the Dexela detector, especially at higher frequencies.
Project description:Positron emission tomography (PET) detectors that use a dual-ended-scintillator readout to measure depth-of-interaction (DOI) must have an accurate DOI function to provide the relationship between DOI and signal ratios to be used for detector calibration and recalibration. In a previous study, the authors used a novel and simple method to accurately and quickly measure DOI function by irradiating the detector with an external uniform flood source; however, as a practical concern, implementing external uniform flood sources in an assembled PET system is technically challenging and expensive. In the current study, therefore, the authors investigated whether the same method could be used to acquire DOI function from scintillator-generated (i.e., internal) radiation. The authors also developed a method for calibrating the energy scale necessary to select the events within the desired energy window.The authors measured the DOI function of a PET detector with lutetium yttrium orthosilicate (LYSO) scintillators. Radiation events originating from the scintillators' internal Lu-176 beta decay were used to measure DOI functions which were then compared with those measured from both an external uniform flood source and an electronically collimated external point source. The authors conducted these studies with several scintillators of differing geometries (1.5 × 1.5 and 2.0 × 2.0 mm(2) cross-section area and 20, 30, and 40 mm length) and various surface finishes (mirror-finishing, saw-cut rough, and other finishes in between), and in a prototype array.All measured results using internal and external radiation sources showed excellent agreement in DOI function measurement. The mean difference among DOI values for all scintillators measured from internal and external radiation sources was less than 1.0 mm for different scintillator geometries and various surface finishes.The internal radioactivity of LYSO scintillators can be used to accurately measure DOI function in PET detectors, regardless of scintillator geometry or surface finish. Because an external radiation source is not needed, this method of DOI function measurement can be practically applied to individual PET detectors as well as assembled systems.
Project description:The advantages of backthinning monolithic active pixel sensors (MAPS) based on complementary metal oxide semiconductor (CMOS) direct electron detectors for electron microscopy have been discussed previously; they include better spatial resolution (modulation transfer function or MTF) and efficiency at all spatial frequencies (detective quantum efficiency or DQE). It was suggested that a 'thin' CMOS detector would have the most outstanding properties [1-3] because of a reduction in the proportion of backscattered electrons. In this paper we show, theoretically (using Monte Carlo simulations of electron trajectories) and experimentally that this is indeed the case. The modulation transfer functions of prototype backthinned CMOS direct electron detectors have been measured at 300keV. At zero spatial frequency, in non-backthinned 700-mum-thick detectors, the backscattered component makes up over 40% of the total signal but, by backthinning to 100, 50 or 35mum, this can be reduced to 25%, 15% and 10%, respectively. For the 35mum backthinned detector, this reduction in backscatter increases the MTF by 40% for spatial frequencies between 0.1 and 1.0 Nyquist. As discussed in the main text, reducing backscattering in backthinned detectors should also improve DQE.
Project description:A new shutterless continuous rotation method using an X-ray complementary metal-oxide semiconductor (CMOS) detector has been developed for high-speed, precise data collection in protein crystallography. The principle of operation and the basic performance of the X-ray CMOS detector (Hamamatsu Photonics KK C10158DK) have been shown to be appropriate to the shutterless continuous rotation method. The data quality of the continuous rotation method is comparable to that of the conventional oscillation method using a CCD detector and, furthermore, the combination with fine ? slicing improves the data accuracy without increasing the data-collection time. The new method is more sensitive to diffraction intensity because of the narrow dynamic range of the CMOS detector. However, the strong diffraction spots were found to be precisely measured by recording them on successive multiple images by selecting an adequate rotation step. The new method has been used to successfully determine three protein structures by multi- and single-wavelength anomalous diffraction phasing and has thereby been proved applicable in protein crystallography. The apparatus and method may become a powerful tool at synchrotron protein crystallography beamlines with important potential across a wide range of X-ray wavelengths.
Project description:Since the 1940s, infrared (IR) detection and imaging at wavelengths in the two atmospheric windows of 3 to 5 and 8 to 14 ?m has been extensively researched. Through several generations, these detectors have undergone considerable developments and have found use in various applications in different fields including military, space science, medicine and engineering. For the most recently proposed generation, these detectors are required to achieve high-speed detection with spectral and polarization selectivity while operating at room temperature. Antenna coupled IR detectors appear to be the most promising candidate to achieve these requirements and has received substantial attention from research in recent years. This paper sets out to present a review of the antenna coupled IR detector family, to explore the main concepts behind the detectors as well as outline their critical and challenging design considerations. In this context, the design of both elements, the antenna and the sensor, will be presented individually followed by the challenging techniques in the impedance matching between both elements. Some hands-on fabrication techniques will then be explored. Finally, a discussion on the coupled IR detector is presented with the aim of providing some useful insights into promising future work.