scholarly journals A modular and compact portable mini-endstation for high-precision, high-speed fixed target serial crystallography at FEL and synchrotron sources

2015 ◽  
Vol 22 (6) ◽  
pp. 1372-1378 ◽  
Author(s):  
Darren A. Sherrell ◽  
Andrew J. Foster ◽  
Lee Hudson ◽  
Brian Nutter ◽  
James O'Hea ◽  
...  
Keyword(s):  
High Precision ◽  
High Speed ◽  
Fixed Target ◽  
Naming Convention ◽  
Large Numbers ◽  
Alignment System ◽  
Specific Position ◽  
Serial Crystallography ◽  
Sample Alignment ◽  
High Quality Sample

The design and implementation of a compact and portable sample alignment system suitable for use at both synchrotron and free-electron laser (FEL) sources and its performance are described. The system provides the ability to quickly and reliably deliver large numbers of samples using the minimum amount of sample possible, through positioning of fixed target arrays into the X-ray beam. The combination of high-precision stages, high-quality sample viewing, a fast controller and a software layer overcome many of the challenges associated with sample alignment. A straightforward interface that minimizes setup and sample changeover time as well as simplifying communication with the stages during the experiment is also described, together with an intuitive naming convention for defining, tracking and locating sample positions. The setup allows the precise delivery of samples in predefined locations to a specific position in space and time, reliably and simply.

Protein crystals IR laser ablated from aqueous solution at high speed retain their diffractive properties: applications in high-speed serial crystallography

2017 ◽  
Vol 50 (6) ◽  
pp. 1773-1781 ◽  
Author(s):  
Eike C. Schulz ◽  
Johannes Kaub ◽  
Frederik Busse ◽  
Pedram Mehrabi ◽  
Henrike M. Müller-Werkmeister ◽  
...  
Keyword(s):  
High Speed ◽  
Vibrational Excitation ◽  
Protein Crystals ◽  
Buffer Solution ◽  
High Repetition ◽  
High Speeds ◽  
Large Numbers ◽  
Heating Effects ◽  
Serial Crystallography ◽  
Jet Injectors

In order to utilize the high repetition rates now available at X-ray free-electron laser sources for serial crystallography, methods must be developed to softly deliver large numbers of individual microcrystals at high repetition rates and high speeds. Picosecond infrared laser (PIRL) pulses, operating under desorption by impulsive vibrational excitation (DIVE) conditions, selectively excite the OH vibrational stretch of water to directly propel the excited volume at high speed with minimized heating effects, nucleation formation or cavitation-induced shock waves, leaving the analytes intact and undamaged. The soft nature and laser-based sampling flexibility provided by the technique make the PIRL system an interesting crystal delivery approach for serial crystallography. This paper demonstrates that protein crystals extracted directly from aqueous buffer solutionviaPIRL-DIVE ablation retain their diffractive properties and can be usefully exploited for structure determination at synchrotron sources. The remaining steps to implement the technology for high-speed serial femtosecond crystallography, such as single-crystal localization, high-speed sampling and synchronization, are described. This proof-of-principle experiment demonstrates the viability of a new laser-based high-speed crystal delivery system without the need for liquid-jet injectors or fixed-target mounting solutions.

scholarly journals On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

IUCrJ ◽  
2019 ◽  
Vol 6 (4) ◽  
pp. 714-728 ◽  
Author(s):  
Julia Lieske ◽  
Maximilian Cerv ◽  
Stefan Kreida ◽  
Dana Komadina ◽  
Janine Fischer ◽  
...  
Keyword(s):  
Binding Studies ◽  
Fixed Target ◽  
X Ray ◽  
Silicon Chips ◽  
Large Numbers ◽  
Human Kinase ◽  
Target Sample ◽  
Crystallization Conditions ◽  
Serial Crystallography ◽  
On Chip

Efficient and reliable sample delivery has remained one of the bottlenecks for serial crystallography experiments. Compared with other methods, fixed-target sample delivery offers the advantage of significantly reduced sample consumption and shorter data collection times owing to higher hit rates. Here, a new method of on-chip crystallization is reported which allows the efficient and reproducible growth of large numbers of protein crystals directly on micro-patterned silicon chips for in-situ serial crystallography experiments. Crystals are grown by sitting-drop vapor diffusion and previously established crystallization conditions can be directly applied. By reducing the number of crystal-handling steps, the method is particularly well suited for sensitive crystal systems. Excessive mother liquor can be efficiently removed from the crystals by blotting, and no sealing of the fixed-target sample holders is required to prevent the crystals from dehydrating. As a consequence, `naked' crystals are obtained on the chip, resulting in very low background scattering levels and making the crystals highly accessible for external manipulation such as the application of ligand solutions. Serial diffraction experiments carried out at cryogenic temperatures at a synchrotron and at room temperature at an X-ray free-electron laser yielded high-quality X-ray structures of the human membrane protein aquaporin 2 and two new ligand-bound structures of thermolysin and the human kinase DRAK2. The results highlight the applicability of the method for future high-throughput on-chip screening of pharmaceutical compounds.

scholarly journals High-speed raster-scanning synchrotron serial microcrystallography with a high-precision piezo-scanner

2018 ◽  
Vol 25 (5) ◽  
pp. 1362-1370 ◽  
Author(s):  
Yuan Gao ◽  
Weihe Xu ◽  
Wuxian Shi ◽  
Alexei Soares ◽  
Jean Jakoncic ◽  
...  
Keyword(s):  
High Precision ◽  
High Speed ◽  
Region Of Interest ◽  
High Dose Rate ◽  
Frame Rate ◽  
Photon Flux ◽  
High Dose ◽  
Raster Scanning ◽  
Dose Rates ◽  
Serial Crystallography

The Frontier Microfocus Macromolecular Crystallography (FMX) beamline at the National Synchrotron Light Source II with its 1 µm beam size and photon flux of 3 × 1012 photons s−1 at a photon energy of 12.66 keV has reached unprecedented dose rates for a structural biology beamline. The high dose rate presents a great advantage for serial microcrystallography in cutting measurement time from hours to minutes. To provide the instrumentation basis for such measurements at the full flux of the FMX beamline, a high-speed, high-precision goniometer based on a unique XYZ piezo positioner has been designed and constructed. The piezo-based goniometer is able to achieve sub-100 nm raster-scanning precision at over 10 grid-linepairs s−1 frequency for fly scans of a 200 µm-wide raster. The performance of the scanner in both laboratory and serial crystallography measurements up to the maximum frame rate of 750 Hz of the Eiger 16M's 4M region-of-interest mode has been verified in this work. This unprecedented experimental speed significantly reduces serial-crystallography data collection time at synchrotrons, allowing utilization of the full brightness of the emerging synchrotron radiation facilities.

scholarly journals Variable step control to improve scanning speed of gene sequencing stage

2021 ◽  
pp. 002029402110022
Author(s):  
Xiaohua Zhou ◽  
Jianbin Zheng ◽  
Xiaoming Wang ◽  
Wenda Niu ◽  
Tongjian Guo
Keyword(s):  
Motion Control ◽  
High Precision ◽  
High Speed ◽  
Stable State ◽  
Gene Sequencing ◽  
Scanning Speed ◽  
Settling Time ◽  
Step Motion ◽  
Variable Step ◽  
Step Control

High-speed scanning is a huge challenge to the motion control of step-scanning gene sequencing stage. The stage should achieve high-precision position stability with minimal settling time for each step. The existing step-scanning scheme usually bases on fixed-step motion control, which has limited means to reduce the time cost of approaching the desired position and keeping high-precision position stability. In this work, we focus on shortening the settling time of stepping motion and propose a novel variable step control method to increase the scanning speed of gene sequencing stage. Specifically, the variable step control stabilizes the stage at any position in a steady-state interval rather than the desired position on each step, so that reduces the settling time. The resulting step-length error is compensated in the next acceleration and deceleration process of stepping to avoid the accumulation of errors. We explicitly described the working process of the step-scanning gene sequencer and designed the PID control structure used in the variable step control for the gene sequencing stage. The simulation was performed to check the performance and stability of the variable step control. Under the conditions of the variable step control where the IMA6000 gene sequencer prototype was evaluated extensively. The experimental results show that the real gene sequencer can step 1.54 mm in 50 ms period, and maintain a high-precision stable state less than 30 nm standard deviation in the following 10 ms period. The proposed method performs well on the gene sequencing stage.

scholarly journals Polyimide mesh-based sample holder with irregular crystal mounting holes for fixed-target serial crystallography

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ki Hyun Nam ◽  
Jihan Kim ◽  
Yunje Cho
Keyword(s):  
Data Collection ◽  
Room Temperature ◽  
Picosecond Laser ◽  
Temperature Structure ◽  
Sample Holder ◽  
Fixed Target ◽  
Random Orientation ◽  
Physical Damage ◽  
Crystal Sample ◽  
Serial Crystallography

AbstractThe serial crystallography (SX) technique enables the determination of the room-temperature structure of a macromolecule while causing minimal radiation damage, as well as the visualization of the molecular dynamics by time-resolved studies. The fixed-target (FT) scanning approach is one method for SX sample delivery that minimizes sample consumption and minimizes physical damage to crystals during data collection. Settling of the crystals on the sample holder in random orientation is important for complete three dimensional data collection. To increase the random orientation of crystals on the sample holder, we developed a polyimide mesh-based sample holder with irregular crystal mounting holes for FT-SX. The polyimide mesh was fabricated using a picosecond laser. Each hole in the polyimide mesh has irregularly shaped holes because of laser thermal damage, which may cause more crystals to settle at random orientations compared to regular shaped sample holders. A crystal sample was spread onto a polyimide-mesh, and a polyimide film was added to both sides to prevent dehydration. Using this sample holder, FT-SX was performed at synchrotron and determined the room-temperature lysozyme structure at 1.65 Å. The polyimide mesh with irregularly shaped holes will allow for expanded applications in sample delivery for FT-SX experiments.

High-speed and high-precision fluorescence-based cell count and viability assays using the Cellaca™ MX high-throughput cell counter

Cytotherapy ◽  
2021 ◽  
Vol 23 (5) ◽  
pp. S97
Author(s):  
J. Bell ◽  
Y. Huang ◽  
S. Yung ◽  
H. Qazi ◽  
C. Hernandez ◽  
...  
Keyword(s):  
Cell Count ◽  
High Precision ◽  
High Throughput ◽  
High Speed ◽  
Cell Counter

Geometrical design of a crystal growth system guided by a machine learning algorithm

CrystEngComm ◽  
2021 ◽  
Author(s):  
Wancheng Yu ◽  
Can Zhu ◽  
Yosuke Tsunooka ◽  
Wei Huang ◽  
Yifan Dang ◽  
...  
Keyword(s):  
Machine Learning ◽  
Crystal Growth ◽  
High Speed ◽  
Learning Algorithm ◽  
Computational Techniques ◽  
Machine Learning Algorithm ◽  
Geometrical Design ◽  
Large Numbers ◽  
Growth System ◽  
Speed Method

This study proposes a new high-speed method for designing crystal growth systems. It is capable of optimizing large numbers of parameters simultaneously which is difficult for traditional experimental and computational techniques.

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