SBIR Phase I: CryoPREP-EM: Time-Resolved Sample Preparation for Single-Particle Cryo-Electron Microscopy (Cryo-EM)

这个小企业创新研究第一阶段项目将评估一种用于低温电子显微镜研究的新型样品制备方法的技术可行性和商业可行性。作为结构生物学领域的一项快速发展的技术，冷冻电镜市场作为一个整体估计每年超过50亿美元，其中5亿美元归因于样品制备领域。尽管冷冻电镜样品制备的整体收入增长率每年超过70%，但在过去十年中，冷冻电镜样品制备仍然是一个尚未看到任何重大技术突破商业化的领域。事实上，据报道，目前的行业解决方案的故障率为95%。通过提供一个实际有效的样品制备系统，这个小企业创新研究（SBIR）第一阶段项目将显著重塑大规模和快速扩张的低温电镜市场，并允许科学家更容易地收集原始状态下的详细蛋白质结构。这种关于细胞和生命本身的分子基础的新知识将用于促进和加速新药物的开发以及疾病的治疗方法，从而从长远来看极大地有益于人类健康。该项目的智能优点是开发了一种低温电镜样品制备装置，该装置建立在应用最先进的微流体射流系统的基础上，可以在几毫秒内在电镜网格上沉积和玻璃化蛋白质溶液，这比目前的方法快至少1000倍。此外，它为科学家提供了控制玻璃化水层厚度的新能力，总的来说，对样品玻璃化过程有更多的控制。有了这种新的样品制备设备，冷冻电镜网格的更大区域将具有完整的蛋白质分子，具有适合结构测定的层厚度，使研究人员能够以高通量的方式在网格制备和筛选方面节省时间和金钱。目前，这项提议的技术还没有经过测试，也没有在任何地方使用。因此，这个SBIR第一阶段项目的首要目标是更好地理解和很好地描述这一拟议创新的技术能力和局限性，在产品商业化之前需要额外的研发工作，预计在五年内。该项目将专注于定义和优化几何和实验设置，以降低技术风险。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This Small Business Innovation Research Phase I project will evaluate the technical feasibility and commercial viability of a novel sample preparation method for cryo-EM (cryogenic electron microscopy) studies. As a rapidly growing technique in the structural biology field, the cryo-EM market as a whole is estimated to be more than $5 billion a year, with $500 million attributed to the sample preparation space. In spite of an overall revenue growth rate of more than 70% per year, cryo-EM sample preparation remains an area that has yet to see any major technological breakthroughs commercialized in the past decade. In fact, the current industry solution is reported to have a failure rate of 95%. By providing a sample preparation system that actually works, this Small Business Innovation Research (SBIR) Phase I project will significantly reshape the massive and rapidly expanding cryo-EM market and allow scientists to more readily collect detailed protein structures in their native state. This new knowledge on the molecular basis of cells and of life itself will be used to facilitate as well as accelerate the development of new pharmaceuticals as well as therapeutic treatments for diseases and thus greatly benefit human health in the long run.The intellectual merit of this project is the development of a cryo-EM sample preparation device built on the application of state-of-the-art microfluidic jet systems to deposit and vitrify protein solution on EM grids in milliseconds, which is at least 1000x faster than the current methods. Furthermore, it offers scientists the novel ability to control the thickness of the vitrified aqueous layer and, overall, more control over the sample vitrification process. With this new sample preparation device, a larger region of the cryo-EM grids will have intact protein molecules with the right layer thickness for structural determination, allowing researchers to save time and money in terms of both grid preparation and screening in a high-throughput manner. Currently, the proposed technology has not been tested nor used anywhere. Therefore, the first and foremost goal of this SBIR Phase I project is to better understand and well characterize the technical capabilities and limitations of this proposed innovation, with additional R&D work needed before product commercialization, which is expected in five years. The project will focus on defining and optimizing both the geometric and experimental setup in order to de-risk the technology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.