Pink-beam serial crystallography

Meents, A.; Wiedorn, Max O.; Šrajer, V.; Henning, Robert; Sarrou, Iosifina; Bergtholdt, J.; Barthelmeß, Miriam; Reinke, P.; Dierksmeyer, D.; Tolstikova, A.; Schaible, S.; Messerschmidt, Marc; Ogata, Craig M.; Kissick, David J.; Taft, Manuel H.; Manstein, Dietmar J.; Lieske, J.; Oberthüer, D.; Fischetti, Robert F.; Chapman, Henry N.

doi:10.1038/s41467-017-01417-3

Cited by 116 publications

(107 citation statements)

References 62 publications

Supporting

Mentioning

106

Contrasting

Order By: Relevance

“…We include in this design a means to reduce the background contribution of Compton scattering from air without having to use a vacuum environment, which would otherwise increase the complexity of the detector and sample preparation. Much of the scattering of the primary beam can be shielded from the detector with a small-diameter thin-walled beampipe that encloses the focused beam and which extends as close as possible to the sample [43].…”

Section: Conceptual Setupmentioning

confidence: 99%

Dose efficient Compton X-ray microscopy

Villanueva-Pérez¹,

Bajt²,

Chapman³

2018

Optica

Self Cite

View full text Add to dashboard Cite

X-ray imaging techniques have proven invaluable to study biological systems at high resolution due to the penetration power and short wavelength of this radiation. In practice, the resolution and sensitivity of current X-ray imaging techniques are not limited by the performance of optics or image-recovery methods but by radiation damage. We propose the use of Compton (inelastic) X-ray scattering for high-resolution cellular imaging and provide a study of a scanning microscope geometry that requires a dose to achieve a given resolution that is three orders of magnitude lower than for coherent (elastic) scattering. We find that the dose per imaging signal is minimized at a photon energy of 64 keV. This corresponds to a short enough wavelength (0.02 nm) to provide nanometer transverse resolution and micrometer depth of field for tomographic imaging of whole cells. The microscope could be implemented at future high-energy and high-brightness synchrotron-radiation facilities to provide images of unsectioned and unlabeled cells in their native conditions at enough detail to bridge the techniques of super-resolution optical microscopy and cryo-electron microscopy. © 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement OCIS codes: (110.7440) X-ray imaging; (340.7460) X-ray microscopy; (170.3880) Medical and biological imaging.

show abstract

Section: Conceptual Setupmentioning

confidence: 99%

Dose efficient Compton X-ray microscopy

Villanueva-Pérez¹,

Bajt²,

Chapman³

2018

Optica

Self Cite

View full text Add to dashboard Cite

show abstract

“…Briefly, crystals can be flowed through quartz capillaries (42) or a microfluidic flow-focusing device (43), or embedded in lipidic cubic phase (44) or other viscous media (45), and then slowly extruded across the X-ray beam. Alternatively, crystals can be presented on fixed targets, such as silicon nitride wafer sandwiches (30), silicon chips (46,47), or Mylar sheet-on-sheet sandwiches (48), and X-ray data collected by raster scanning. As an in-between the injection and fixed-target approaches, tape drives carrying microcrystals are yet another way of delivering crystalline samples to the X-ray beam (49,50).…”

mentioning

confidence: 99%

Radiation damage and dose limits in serial synchrotron crystallography at cryo- and room temperatures

Mora

Coquelle

Bury

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

103

View full text Add to dashboard Cite

Radiation damage limits the accuracy of macromolecular structures in X-ray crystallography. Cryogenic (cryo-) cooling reduces the global radiation damage rate and, therefore, became the method of choice over the past decades. The recent advent of serial crystallography, which spreads the absorbed energy over many crystals, thereby reducing damage, has rendered room temperature (RT) data collection more practical and also extendable to microcrystals, both enabling and requiring the study of specific and global radiation damage at RT. Here, we performed sequential serial raster-scanning crystallography using a microfocused synchrotron beam that allowed for the collection of two series of 40 and 90 full datasets at 2- and 1.9-Å resolution at a dose rate of 40.3 MGy/s on hen egg white lysozyme (HEWL) crystals at RT and cryotemperature, respectively. The diffraction intensity halved its initial value at average doses (D1/2) of 0.57 and 15.3 MGy at RT and 100 K, respectively. Specific radiation damage at RT was observed at disulfide bonds but not at acidic residues, increasing and then apparently reversing, a peculiar behavior that can be modeled by accounting for differential diffraction intensity decay due to the nonuniform illumination by the X-ray beam. Specific damage to disulfide bonds is evident early on at RT and proceeds at a fivefold higher rate than global damage. The decay modeling suggests it is advisable not to exceed a dose of 0.38 MGy per dataset in static and time-resolved synchrotron crystallography experiments at RT. This rough yardstick might change for proteins other than HEWL and at resolutions other than 2 Å.

show abstract

“…A photon energy of 7.15 keV, pulse length of 10 fs and a repetition rate of 120 Hz was used throughout the experiments. DFFN nozzle (design 8) was used for sample delivery on the RoadRunner system 70 adapted for liquid jets with the capillary beamstop 41 and a helium enclosure. The Cornell-SLAC pixel-array detector 71 (CSPAD) was used for data collection.…”

Section: Methodsmentioning

confidence: 99%

“…This confirms that the low-speed jet injection approach with our 3D-printed DFFN can reduce sample consumption rates when determining X-ray structure void of conventional X-ray damage at low-repetition XFELs. Since the DFFN can also inject larger crystals, we anticipate this injector to also be suitable for low-speed jet injection using microfocus polychromatic synchrotron radiation 41 . In particular in combination with mixing or pump-probe approaches this may greatly extend accessibility to time-resolved serial crystallography.…”

Section: Multi-orifice Nozzle As a Versatile Low Consumption Injectormentioning

confidence: 99%

Ultracompact 3D microfluidics for time-resolved structural biology

et al. 2020

Self Cite

View full text Add to dashboard Cite

To advance microfluidic integration, we present the use of two-photon additive manufacturing to fold 2D channel layouts into compact free-form 3D fluidic circuits with nanometer precision. We demonstrate this technique by tailoring microfluidic nozzles and mixers for time-resolved structural biology at X-ray free-electron lasers (XFELs). We achieve submicron jets with speeds exceeding 160 m s −1 , which allows for the use of megahertz XFEL repetition rates. By integrating an additional orifice, we implement a low consumption flowfocusing nozzle, which is validated by solving a hemoglobin structure. Also, aberration-free in operando X-ray microtomography is introduced to study efficient equivolumetric millisecond mixing in channels with 3D features integrated into the nozzle. Such devices can be printed in minutes by locally adjusting print resolution during fabrication. This technology has the potential to permit ultracompact devices and performance improvements through 3D flow optimization in all fields of microfluidic engineering.

show abstract

Pink-beam serial crystallography

Cited by 116 publications

References 62 publications

Dose efficient Compton X-ray microscopy

Dose efficient Compton X-ray microscopy

Radiation damage and dose limits in serial synchrotron crystallography at cryo- and room temperatures

Ultracompact 3D microfluidics for time-resolved structural biology

Contact Info

Product

Resources

About