Plug-and-play polymer microfluidic chips for hydrated, room temperature, fixed-target serial crystallography.
Plug-and-play polymer microfluidic chips for hydrated, room temperature, fixed-target serial crystallography.
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DOI：
10.1039/d1lc00810b
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发表时间：
2021-12-07
期刊：
影响因子：
6.1
通讯作者：
Kuhl TL
中科院分区：
文献类型：
--
作者：
Gilbile D;Shelby ML;Lyubimov AY;Wierman JL;Monteiro DCF;Cohen AE;Russi S;Coleman MA;Frank M;Kuhl TL
连续X射线结晶学(SX)的实践依赖于高效、连续地输送水合蛋白质晶体，同时将背景散射降至最低。在两种主要类型的样品输送装置中，固定靶标装置比广泛采用的喷射注射器具有几个优势，包括：样品消耗更低、无堵塞输送，以及能够控制芯片上的晶体密度以提高命中率。在这里，我们介绍了我们开发的多功能、廉价和坚固耐用的聚合物微流控芯片，用于同步加速器和X射线自由电子激光(XFELs)的常规和可靠的室温连续测量。我们的设计包括可调至最小散射背景的高度X射线透明的封闭薄膜层、可调整以匹配晶体大小的自适应样品流动层，以及兼容栅格扫描和基于旋转的串行数据采集的大样品区域。这种光学透明芯片既可用于原位蛋白质结晶(以避免晶体处理)，也可用于晶体浆料装载，所制备的样品在潮湿的环境中可稳定数周，在环境条件下可稳定数小时。在微聚焦同步加速器光束线(SSRL，光束线12-1)上使用使用多晶体旋转数据收集方法的连续振荡结晶学来对芯片的性能进行基准测试。从五种不同的蛋白质--蛋清溶菌酶、索马菌素、牛肝过氧化氢酶、刀豆蛋白A(VI型)和SARS-CoV-2非结构蛋白NSP5中收集了高分辨结构(1.3-2.7ä)。总体而言，我们的模块化制造方法能够精确控制X射线光束路径中的材料横截面，并便于芯片适应不同的样品和光束线要求，以便在室温下进行用户友好、直接的衍射测量。这项工作介绍了我们开发的多功能、廉价和坚固耐用的聚合物微流控芯片，用于常规和可靠的室温系列X射线结晶学测量。
The practice of serial X-ray crystallography (SX) depends on efficient, continuous delivery of hydrated protein crystals while minimizing background scattering. Of the two major types of sample delivery devices, fixed-target devices offer several advantages over widely adopted jet injectors, including: lower sample consumption, clog-free delivery, and the ability to control on-chip crystal density to improve hit rates. Here we present our development of versatile, inexpensive, and robust polymer microfluidic chips for routine and reliable room-temperature serial measurements at both synchrotrons and X-ray free electron lasers (XFELs). Our design includes highly X-ray-transparent enclosing thin film layers tuned to minimize scatter background, adaptable sample flow layers tuned to match crystal size, and a large sample area compatible with both raster scanning and rotation based serial data collection. The optically transparent chips can be used both for in situ protein crystallization (to eliminate crystal handling) or crystal slurry loading, with prepared samples stable for weeks in a humidified environment and for several hours in ambient conditions. Serial oscillation crystallography, using a multi-crystal rotational data collection approach, at a microfocus synchrotron beamline (SSRL, beamline 12–1) was used to benchmark the performance of the chips. High-resolution structures (1.3–2.7 Å) were collected from five different proteins- hen egg white lysozyme, thaumatin, bovine liver catalase, concanavalin-A (type VI), and SARS-CoV-2 nonstructural protein NSP5. Overall, our modular fabrication approach enables precise control over the cross-section of materials in the X-ray beam path and facilitates chip adaption to different sample and beamline requirements for user-friendly, straightforward diffraction measurements at room temperature. This work presents our development of versatile, inexpensive, and robust polymer microfluidic chips for routine and reliable room-temperature serial X-ray crystallography measurements.
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影响因子：
48
作者：
Fuller FD;Gul S;Chatterjee R;Burgie ES;Young ID;Lebrette H;Srinivas V;Brewster AS;Michels-Clark T;Clinger JA;Andi B;Ibrahim M;Pastor E;de Lichtenberg C;Hussein R;Pollock CJ;Zhang M;Stan CA;Kroll T;Fransson T;Weninger C;Kubin M;Aller P;Lassalle L;Bräuer P;Miller MD;Amin M;Koroidov S;Roessler CG;Allaire M;Sierra RG;Docker PT;Glownia JM;Nelson S;Koglin JE;Zhu D;Chollet M;Song S;Lemke H;Liang M;Sokaras D;Alonso-Mori R;Zouni A;Messinger J;Bergmann U;Boal AK;Bollinger JM Jr;Krebs C;Högbom M;Phillips GN Jr;Vierstra RD;Sauter NK;Orville AM;Kern J;Yachandra VK;Yano J
通讯作者：
Yano J
DOI：
10.1107/s2059798315020847
发表时间：
2016-01
期刊：
Acta crystallographica. Section D, Structural biology
影响因子：
--
作者：
Baxter EL;Aguila L;Alonso-Mori R;Barnes CO;Bonagura CA;Brehmer W;Brunger AT;Calero G;Caradoc-Davies TT;Chatterjee R;Degrado WF;Fraser JS;Ibrahim M;Kern J;Kobilka BK;Kruse AC;Larsson KM;Lemke HT;Lyubimov AY;Manglik A;McPhillips SE;Norgren E;Pang SS;Soltis SM;Song J;Thomaston J;Tsai Y;Weis WI;Woldeyes RA;Yachandra V;Yano J;Zouni A;Cohen AE
通讯作者：
Cohen AE
DOI：
10.1126/science.1217737
发表时间：
2012-07-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Boutet S;Lomb L;Williams GJ;Barends TR;Aquila A;Doak RB;Weierstall U;DePonte DP;Steinbrener J;Shoeman RL;Messerschmidt M;Barty A;White TA;Kassemeyer S;Kirian RA;Seibert MM;Montanez PA;Kenney C;Herbst R;Hart P;Pines J;Haller G;Gruner SM;Philipp HT;Tate MW;Hromalik M;Koerner LJ;van Bakel N;Morse J;Ghonsalves W;Arnlund D;Bogan MJ;Caleman C;Fromme R;Hampton CY;Hunter MS;Johansson LC;Katona G;Kupitz C;Liang M;Martin AV;Nass K;Redecke L;Stellato F;Timneanu N;Wang D;Zatsepin NA;Schafer D;Defever J;Neutze R;Fromme P;Spence JC;Chapman HN;Schlichting I
通讯作者：
Schlichting I
DOI：
10.1073/pnas.1418733111
发表时间：
2014-12-02
影响因子：
11.1
作者：
Cohen, Aina E.;Soltis, S. Michael;Hodgson, Keith O.
通讯作者：
Hodgson, Keith O.
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K