Electron imaging with Medipix2 hybrid pixel detector

McMullan, G.; Cattermole, D.M.; Chen, S.; Henderson, Richard A.; Llopart, X.; Summerfield, C.; Tlustos, L.; Faruqi, A.R.

doi:10.1016/j.ultramic.2006.10.005

Cited by 94 publications

(98 citation statements)

References 31 publications

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“…In silicon $3.6 eV is required to produce one electron-hole pair; therefore, a 200 keV electron can produce $55 000 pairs. This means that the dynamic range of the electronic noise accounts for less than 1.6% of the total deposited energy per electron incident and even less when there is more than one electron incident per clock cycle (McMullan et al, 2007;Plackett et al, 2010).…”

Section: Measuring Electrons Using a Medipix2 Detectormentioning

confidence: 99%

A Medipix quantum area detector allows rotation electron diffraction data collection from submicrometre three-dimensional protein crystals

Nederlof

Genderen

et al. 2013

Acta Crystallogr D Biol Crystallogr

179

183

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When protein crystals are submicrometre-sized, X-ray radiation damage precludes conventional diffraction data collection. For crystals that are of the order of 100 nm in size, at best only single-shot diffraction patterns can be collected and rotation data collection has not been possible, irrespective of the diffraction technique used. Here, it is shown that at a very low electron dose (at most 0.1 e À Å À2 ), a Medipix2 quantum area detector is sufficiently sensitive to allow the collection of a 30-frame rotation series of 200 keV electrondiffraction data from a single $100 nm thick protein crystal. A highly parallel 200 keV electron beam ( = 0.025 Å ) allowed observation of the curvature of the Ewald sphere at low resolution, indicating a combined mosaic spread/beam divergence of at most 0.4 . This result shows that volumes of crystal with low mosaicity can be pinpointed in electron diffraction. It is also shown that strategies and data-analysis software (MOSFLM and SCALA) from X-ray protein crystallography can be used in principle for analysing electron-diffraction data from three-dimensional nanocrystals of proteins.

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Section: Measuring Electrons Using a Medipix2 Detectormentioning

confidence: 99%

A Medipix quantum area detector allows rotation electron diffraction data collection from submicrometre three-dimensional protein crystals

Nederlof

Genderen

et al. 2013

Acta Crystallogr D Biol Crystallogr

179

183

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“…A quantitative comparison between image plates and a Medipix hybrid pixel detector indicated a substantial improvement to be offered by the latter (Georgieva et al, 2011;Nederlof et al, 2013). Hybrid pixel detectors such as Medipix, Timepix and EIGER (Llopart et al, 2002Johnson et al, 2012) are well suited for measuring high-energy electrons (McMullan et al, 2007), and can overcome difficulties in detecting weak peaks, for example for CCD and CMOS detectors (Hattne et al, 2016;Rodriguez & Gonen, 2016). …”

Section: Discussionmentioning

confidence: 99%

“…For 200 and 300 keV electrons the potential scattering distances are approximately 225 and 450 mm, respectively (McMullan et al, 2007(McMullan et al, , 2009Faruqi & McMullan, 2011). For the prototype, we used a 300 mm sensitive silicon layer.…”

Section: Data Acquisitionmentioning

confidence: 99%

“…However, because of the perpendicular impact of a 300 keV incident electron on the detector, on average the first pixel and the last pixel of its track receive the highest deposited dose. This means that at the energy threshold used for each pixel ($60 keV), the electron is counted once (70%) or twice (30%) (McMullan et al, 2007(McMullan et al, , 2009Faruqi & McMullan, 2011). This means that the Bragg spot is spread out over a larger area.…”

Section: Data Acquisitionmentioning

confidence: 99%

“…Since electron diffraction data can have spikes of high intensity at low resolution and in Bragg peaks, monolithic detectors are currently not used for measuring electron diffraction data. However, in hybrid pixel detectors the high-energy electrons are stopped by the silicon sensor layer that is bump-bonded to the counting and integration electronics (McMullan et al, 2007(McMullan et al, , 2009Faruqi & McMullan, 2011). The integrating electronics of CMOS detectors can be shielded by a phosphor, at the expense of an increased point spread.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Protein structure determination by electron diffraction using a single three-dimensional nanocrystal

Clabbers

Genderen

Wan

et al. 2017

Acta Crystallogr D Struct Biol

100

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Three-dimensional nanometre-sized crystals of macromolecules currently resist structure elucidation by single-crystal X-ray crystallography. Here, a single nanocrystal with a diffracting volume of only 0.14 mm 3 , i.e. no more than 6 Â 10 5 unit cells, provided sufficient information to determine the structure of a rare dimeric polymorph of hen egg-white lysozyme by electron crystallography. This is at least an order of magnitude smaller than was previously possible. The molecular-replacement solution, based on a monomeric polyalanine model, provided sufficient phasing power to show side-chain density, and automated model building was used to reconstruct the side chains. Diffraction data were acquired using the rotation method with parallel beam diffraction on a Titan Krios transmission electron microscope equipped with a novel in-house-designed 1024 Â 1024 pixel Timepix hybrid pixel detector for low-dose diffraction data collection. Favourable detector characteristics include the ability to accurately discriminate single high-energy electrons from X-rays and count them, fast readout to finely sample reciprocal space and a high dynamic range. This work, together with other recent milestones, suggests that electron crystallography can provide an attractive alternative in determining biological structures.

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Membrane Protein Structure Determination by Electron Cryo‐Microscopy

Tate

Rubinstein

2007

Biophysical Analysis of Membrane Proteins

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Electron imaging with Medipix2 hybrid pixel detector

Cited by 94 publications

References 31 publications

A Medipix quantum area detector allows rotation electron diffraction data collection from submicrometre three-dimensional protein crystals

A Medipix quantum area detector allows rotation electron diffraction data collection from submicrometre three-dimensional protein crystals

Protein structure determination by electron diffraction using a single three-dimensional nanocrystal

Membrane Protein Structure Determination by Electron Cryo‐Microscopy

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