Hyperquenching Cryocooler for Biomolecular X-ray Crystallography

描述（由申请人提供）：该项目的总体目标是设计一种紧凑的冷冻冷却装置，用于快速，可重复和可靠的冷冻保存x射线冷冻成像的蛋白质和病毒晶体。x射线晶体学是测定蛋白质、病毒、核酸和生物分子复合物分子结构的最强大和广泛使用的工具。在液氮中快速冷却晶体并在T=100 K下收集数据简化了存储，运输和处理，并显着增加了可以从每个晶体中获得的x射线数据量。然而，冷却过程本身会破坏晶体。许多重要的目标，包括病毒、膜蛋白和生物分子复合物，都很难成功地冷却。此外，目前的冷却方法耗时且手工完成，导致冷却结果和衍射质量的巨大变化。所提出的设备将提供可靠和可重复的冷却速率高达目前最佳实践的100倍。这将消除结晶冰在冷却过程中的形成，并大大减少所需的冷冻保护剂浓度。本项目将侧重于设计设备，开发商业原型，评估原型的性能。

DESCRIPTION (provided by applicant): The overall goal for this project is to design a compact cryocooling device for fast, reproducible and reliable cryopreservation of protein and virus crystals for X-ray cryocrystallography. X-ray crystallography is the most powerful and widely used tool for determining the molecular structure of proteins, viruses, nucleic acids and biomolecular complexes. Flash-cooling crystals in liquid nitrogen and collecting data at T=100 K simplifies storage, transport and handling, and dramatically increases the amount of X-ray data that can be obtained from each crystal. However, the cooling process itself damages crystals. Many important targets including viruses, membrane proteins and biomolecular complexes can be very difficult to successfully cool. Moreover, current cooling methodologies, which are time-consuming and done by hand, lead to large variability in cooling outcomes and diffraction quality. The proposed device will provide reliable and reproducible cooling at rates up to 100 times greater than in current best practice. This will eliminate crystalline ice formation during cooling and dramatically reduce required cryoprotectant concentrations. This project will focus on designing the device, developing a commercial prototype, evaluating the performance of prototype. PUBLIC HEALTH RELEVANCE: This project will benefit public health by developing an improved method of conducting X-ray Cryocrystallography. This technique is vital for basic biomedical research and improving its accuracy and efficacy will have broad-scale impacts on developing new drugs and treatments for diseases.