Droplet microfluidics for time-resolved serial crystallography

被引：1

作者：

Stubbs, Jack ^{[1
,2
]}

Hornsey, Theo ^{[1
]}

Hanrahan, Niall ^{[3
,4
]}

Esteban, Luis Blay ^{[5
]}

Bolton, Rachel ^{[1
,2
]}

Maly, Martin ^{[1
,4
]}

Basu, Shibom ^{[6
]}

Orlans, Julien ^{[7
]}

De Sanctis, Daniele ^{[7
]}

Shim, Jung-Uk ^{[8
]}

Stewart, Patrick D. Shaw ^{[9
]}

Orville, Allen M. ^{[2
,10
]}

Tews, Ivo ^{[1
,4
]}

West, Jonathan ^{[4
,11
]}

机构：

[1] Univ Southampton, Sch Biol Sci, Fac Environm & Life Sci, Southampton SO17 1BJ, England

[2] Harwell Sci & Innovat Campus, Diamond Light Source, Didcot, Oxon, England

[3] Univ South ampton, Sch Chem, Fac Engn & Phys Sci, Southampton SO17 1BJ, England

[4] Univ Southampton, Inst Life Sci, Southampton SO17 1BJ, England

[5] Univ Carlemany, Avenida Verge De Canolich 47,, Principat Dandorra AD600, Spain

[6] European Mol Biol Lab, Grenoble Outstat,71 Ave Martyrs, F-38042 Grenoble 9, CS 90181, France

[7] European Synchrotron Radiat Facil ESRF, 71 Ave Martyrs, F-38042 Grenoble 9, France

[8] Univ Leeds, Fac Engn & Phys Sci, Leeds LS2 9JT, England

[9] Douglas Instruments Ltd, Hungerford RG17 7HD, England

[10] Harwell Sci & Innovat Campus, Res Complex Harwell, Didcot OX11 0FA, Oxon, England

[11] Univ Southampton, Canc Sci, Fac Med, Southampton SO17 1BJ, England

来源：

IUCRJ | 2024年 / 11卷

基金：

英国惠康基金; 英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会; 英国科学技术设施理事会;

关键词：

droplet microfluidics; crystal miniaturization; micromixing; time-resolved serial crystallography; MACROMOLECULAR CRYSTALLOGRAPHY; PROTEIN CRYSTALLIZATION; ROOM-TEMPERATURE; ON-DEMAND; CHIP; INTERMEDIATE; NUCLEATION; INTERFACE; GROWTH;

D O I：

10.1107/S2052252524001799

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3 mu m crystals of lysozyme and 2 mu m crystals of Pdx1, an Arabidopsis enzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of <2 ms were achieved. Droplet microfluidics for crystal size engineering and rapid micromixing can be utilized to advance time-resolved serial crystallography.

引用

页码：237 / 248

页数：12

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