Lipidico Injection Protocol for Serial Crystallography Measurements at the Australian Synchrotron

被引:4
作者
Berntsen, Peter [1 ]
Sharma, Rama [1 ]
Kusel, Michael [2 ]
Abbey, Brian [1 ]
Darmanin, Connie [1 ]
机构
[1] La Trobe Univ, Australian Res Council Ctr Excellence Adv Mol Ima, La Trobe Inst Mol Sci, Dept Chem & Phys, Melbourne, Vic 3086, Australia
[2] Kusel Design, Melbourne, Vic 3042, Australia
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2020年 / 163期
基金
澳大利亚研究理事会;
关键词
FEMTOSECOND CRYSTALLOGRAPHY; MILLISECOND CRYSTALLOGRAPHY; CUBIC PHASE; MICROCRYSTALS;
D O I
10.3791/61650
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A facility for performing serial crystallography measurements has been developed at the Australian synchrotron. This facility incorporates a purpose built high viscous injector, Lipidico, as part of the macromolecular crystallography (MX2) beamline to measure large numbers of small crystals at room temperature. The goal of this technique is to enable crystals to be grown/transferred to glass syringes to be used directly in the injector for serial crystallography data collection. The advantages of this injector include the ability to respond rapidly to changes in the flow rate without interruption of the stream. Several limitations for this high viscosity injector (HVI) exist which include a restriction on the allowed sample viscosities to >10 Pa.s. Stream stability can also potentially be an issue depending on the specific properties of the sample. A detailed protocol for how to set up samples and operate the injector for serial crystallography measurements at the Australian synchrotron is presented here. The method demonstrates preparation of the sample, including the transfer of lysozyme crystals into a high viscous media (silicone grease), and the operation of the injector for data collection at MX2.
引用
收藏
页码:1 / 15
页数:15
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