A.R. Faruqi

Detective quantum efficiency of electron area detectors in electron microscopy.

Recent progress in detector design has created the need for a careful side-by-side comparison of the modulation transfer function (MTF) and resolution-dependent detective quantum efficiency (DQE) of existing electron detectors with those of detectors based on new technology. We present MTF and DQE measurements for four types of detector: Kodak SO-163 film, TVIPS 224 charge coupled device (CCD) detector, the Medipix2 hybrid pixel detector, and an experimental direct electron monolithic active pixel sensor (MAPS) detector. Film and CCD performance was measured at 120 and 300 keV, while results are presented for the Medipix2 at 120 keV and for the MAPS detector at 300 keV. In the case of film, the effects of electron backscattering from both the holder and the plastic support have been investigated. We also show that part of the response of the emulsion in film comes from light generated in the plastic support. Computer simulations of film and the MAPS detector have been carried out and show good agreement with experiment. The agreement enables us to conclude that the DQE of a backthinned direct electron MAPS detector is likely to be equal to, or better than, that of film at 300 keV.

Comparison of optimal performance at 300 keV of three direct electron detectors for use in low dose electron microscopy

Low dose electron imaging applications such as electron cryo-microscopy are now benefitting from the improved performance and flexibility of recently introduced electron imaging detectors in which electrons are directly incident on backthinned CMOS sensors. There are currently three commercially available detectors of this type: the Direct Electron DE-20, the FEI Falcon II and the Gatan K2 Summit. These have different characteristics and so it is important to compare their imaging properties carefully with a view to optimise how each is used. Results at 300 keV for both the modulation transfer function (MTF) and the detective quantum efficiency (DQE) are presented. Of these, the DQE is the most important in the study of radiation sensitive samples where detector performance is crucial. We find that all three detectors have a better DQE than film. The K2 Summit has the best DQE at low spatial frequencies but with increasing spatial frequency its DQE falls below that of the Falcon II.

Experimental observation of the improvement in MTF from backthinning a CMOS direct electron detector

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 300 keV. At zero spatial frequency, in non-backthinned 700-μm-thick detectors, the backscattered component makes up over 40% of the total signal but, by backthinning to 100, 50 or 35 μm, this can be reduced to 25%, 15% and 10%, respectively. For the 35 μm 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.

Enhanced imaging in low dose electron microscopy using electron counting

We compare the direct electron imaging performance at 120 keV of a monolithic active pixel sensor (MAPS) operated in a conventional integrating mode with the performance obtained when operated in a single event counting mode. For the combination of sensor and incident electron energy used here, we propose a heuristic approach with which to process the single event images in which each event is renormalised to have an integrated weight of unity. Using this approach we find enhancements in the Nyquist frequency modulation transfer function (MTF) and detective quantum efficiency (DQE) over the corresponding integrating mode values by factors of 8 and 3, respectively.

Images of paraffin monolayer crystals with perfect contrast: Minimization of beam-induced specimen motion

Quantitative analysis of electron microscope images of organic and biological two-dimensional crystals has previously shown that the absolute contrast reached only a fraction of that expected theoretically from the electron diffraction amplitudes. The accepted explanation for this is that irradiation of the specimen causes beam-induced charging or movement, which in turn causes blurring of the image due to image or specimen movement. In this paper, we used three different approaches to try to overcome this image-blurring problem in monolayer crystals of paraffin. Our first approach was to use an extreme form of spotscan imaging, in which a single image was assembled on film by the successive illumination of up to 50,000 spots, each of a diameter of around 7 nm. The second approach was to use the Medipix II detector with its zero-noise readout to assemble a time-sliced series of images of the same area in which each frame from a movie with up to 400 frames had an exposure of only 500 electrons. In the third approach, we simply used a much thicker carbon support film to increase the physical strength and conductivity of the support. Surprisingly, the first two methods involving dose fractionation in space or time produced only partial improvements in contrast whereas the third approach produced many virtually perfect images, where the absolute contrast predicted from the electron diffraction amplitudes was observed in the images. We conclude that it is possible to obtain consistently almost perfect images of beam-sensitive specimens if they are attached to an appropriately strong and conductive support; however great care is needed in practice and the problem remains of how to best image ice-embedded biological structures in the absence of a strong, conductive support film.

Cooled CCD detector with tapered fibre optics for recording electron diffraction patterns

The construction and evaluation of a cooled CCD detector for 120 keV electrons using a tapered fibre optics is described. It has been used for accurate recording of electron diffraction data, for which it is a nearly perfect detector. The results suggest that the degree of tapering used could be increased considerably without loss of sensitivity, and that such an instrument might then also become a nearly perfect detector for image recording where, at present, the point spread function still limits the performance.

Evaluation of a hybrid pixel detector for electron microscopy

We describe the application of a silicon hybrid pixel detector, containing 64 by 64 pixels, each 170 microm(2), in electron microscopy. The device offers improved resolution compared to CCDs along with faster and noiseless readout. Evaluation of the detector, carried out on a 120 kV electron microscope, demonstrates the potential of the device.

A high frame rate, 16 million pixels, radiation hard CMOS sensor

CMOS sensors provide the possibility of designing detectors for a large variety of applications with all the benefits and flexibility of the widely used CMOS process. In this paper we describe a novel CMOS sensor designed for transmission electron microscopy. The overall design consists of a large 61 × 63 mm2 silicon area containing 16 million pixels arranged in a 4K × 4K array, with radiation hard geometry. All this is combined with a very fast readout, the possibility of region of interest (ROI) readout, pixel binning with consequent frame rate increase and a dynamic range close to 12 bits. The high frame rate has been achieved using 32 parallel analogue outputs each one operating at up to 20 MHz. Binning of pixels can be controlled externally and the flexibility of the design allows several possibilities, such as 2 × 2 or 4 × 4 binning. Other binning configurations where the number of rows and the number of columns are not equal, such as 2 × 1 or 2 × 4, are also possible. Having control of the CMOS design allowed us to optimise the pixel design, in particular with regard to its radiation hardness, and to make optimum choices in the design of other regions of the final sensor. An early prototype was also designed with a variety of geometries in order to optimise the readout structure and these are presented. The sensor was manufactured in a 0.35 μm standard CMOS process.

Prospects for hybrid pixel detectors in electron microscopy

Abstract The current status of CCD-based detectors for cryo-electron microscopy of membrane and other proteins is described briefly, highlighting the strengths and weaknesses of the technique. Over the past few years CCD detectors have been used extensively in electron crystallography of membrane proteins, and in particular, in the study of the molecular transitions which take place during the photo-cycle of the light-driven proton pump bacteriorhodopsin . Direct-detection methods, which avoid the intermediate stages of converting the electron energy into light, offer the possibility of improved spatial resolution compared to CCD detectors; in addition, photon counting and noise-free readout should improve the signal-to-noise ratio.

Development and application of multiwire detectors in biological X-ray studies

Abstract The development of a number of multiwire detectors is described for use in structural studies of biological specimen. A few examples taken from small angle scattering and diffraction and crystallography are used to illustrate the nature of the applications. As most of the detector work was originally directed at time-resolved studies of skeletal muscle, it forms the main part of the paper.