Over the past two decades, serial X-ray crystallography has enabled the structure determination of a wide range of proteins. With the advent of X-ray free-electron lasers (XFELs), ever-smaller crystals have yielded high-resolution diffraction and structure determination. A crucial need to continue advancement is the efficient delivery of fragile and micrometre-sized crystals to the X-ray beam intersection. This paper presents an improved design of an all-polymer microfluidic `chip' for room-temperature fixed-target serial crystallography that can be tailored to broadly meet the needs of users at either synchrotron or XFEL light sources. The chips are designed to be customized around different types of crystals and offer users a friendly, quick, convenient, ultra-low-cost and robust sample-delivery platform. Compared with the previous iteration of the chip [Gilbile et al. (2021), Lab Chip, 21, 4831–4845], the new design eliminates cleanroom fabrication. It has a larger imaging area to volume, while maintaining crystal hydration stability for both in situ crystallization or direct crystal slurry loading. Crystals of two model proteins, lysozyme and thaumatin, were used to validate the effectiveness of the design at both synchrotron (lysozyme and thaumatin) and XFEL (lysozyme only) facilities, yielding complete data sets with resolutions of 1.42, 1.48 and 1.70 Å, respectively. Overall, the improved chip design, ease of fabrication and high modifiability create a powerful, all-around sample-delivery tool that structural biologists can quickly adopt, especially in cases of limited sample volume and small, fragile crystals.

中文翻译：

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一种用户友好的即插即用的基于环烯烃共聚物的微流控芯片，用于室温、固定目标串行晶体学

在过去的二十年中，连续 X 射线晶体学已经能够确定多种蛋白质的结构。随着 X 射线自由电子激光器 (XFEL) 的出现，越来越小的晶体已经可以实现高分辨率衍射和结构测定。继续进步的一个关键需求是将易碎的微米级晶体有效地输送到 X 射线束交叉点。本文提出了一种用于室温固定目标串行晶体学的全聚合物微流体“芯片”的改进设计，该芯片可以进行定制，以广泛满足用户对同步加速器或 XFEL 光源的需求。这些芯片旨在围绕不同类型的晶体进行定制，为用户提供友好、快速、方便、超低成本和强大的样品交付平台。与芯片的先前迭代相比[Gilbile等人。(2021), Lab Chip , 21 , 4831–4845]，新设计消除了洁净室制造。它具有更大的成像面积与体积，同时保持原位结晶或直接晶体浆料加载的晶体水合稳定性。使用两种模型蛋白（溶菌酶和索马甜）的晶体在同步加速器（溶菌酶和索马甜）和 XFEL（仅溶菌酶）设施上验证设计的有效性，产生分辨率分别为 1.42、1.48 和 1.70 Å 的完整数据集。总体而言，改进的芯片设计、易于制造和高可修改性创造了结构生物学家可以快速采用的强大、全方位的样品交付工具，特别是在样品量有限和小而脆弱的晶体的情况下。