SBIR Phase II: Real-time Active Image Stabilization for Microscopy

这个小型企业创新研究(SBIR)第二阶段项目将开发一种集成系统并将其商业化，以积极地将光学显微镜稳定到当今尖端成像方法所需的精度。生物科学中的显微镜在几个方面正在经历根本性的进步。“超分辨率”(SR)技术绕过了曾经被认为无法克服的衍射分辨率限制，并承诺能够在分子水平上对细胞生物学的结构和过程进行成像。这将带来对细胞内部工作原理的深刻理解。然而，在广泛使用SR技术的道路上仍然存在重大的相互关联的障碍：(1)它们在技术上具有挑战性，(2)实施起来成本高昂；(3)它们对其设计不能满足的显微镜平台提出了物理要求。这些要求中最重要的是，SR方法需要控制生物样品的移动和纳米级精度的显微镜系统的稳定性。这个商业化的集成系统就是专门为解决这些问题和消除这些障碍而设计的。它使用一个三轴、压电驱动的纳米定位平台来控制样品的运动，并以图像作为这种稳定性的参考点来积极地维持系统的稳定性。该项目的更广泛的影响/商业潜力在于使SR方法对工作的生物学家常规有用。这些“改变游戏规则”的工具将促进我们对疾病病理的分子基础的理解，并使针对其治疗的更严格的方法成为可能。新的见解将从分子病毒学和开发更安全、更有效的疫苗，到神经元信号和学习和记忆的分子机制。事实上，很难想象细胞生物学领域不会受到这些新兴的SR技术的影响。这些方法的先驱之一将它们比作哈勃望远镜：它们使人们能够看到他们以前根本看不到的东西。这个类比更进一步：由于常规使用的技术和经济障碍，目前只有一个哈勃望远镜，而且目前具有SR能力的成像系统非常少。虽然SR方法以一种尖锐的方式暴露了显微镜的物理局限性，但它们的稳定性和图像获取要求并不是唯一的。因此，这一商业系统将有更广泛的用途：它还将在纳米级实现焦点稳定和分子跟踪，用于任何长期成像实验。

This Small Business Innovation Research (SBIR) Phase II project will develop and commercialize an integrated system to actively stabilize an optical microscope to the precision required by today's cutting-edge imaging methods. Microscopy in the biological sciences is undergoing radical advancement on several fronts. "Super-Resolution" (SR) techniques circumvent the diffraction limit on resolution once thought to be insurmountable, and promise the ability to image the structures and processes of cell biology at the molecular level. This will usher in profound advancements in the understanding of the inner-workings of the cell. However, significant interrelated barriers remain in the path towards widespread use of SR techniques: (1) they are technically challenging and (2) expensive to implement; and (3) they place physical demands on the microscope platform it was not designed to meet. Foremost of these demands is that SR methods require control over the movement of the biological sample and the stability of the microscope system with nanometer precision. This commercialized integrated system is designed specifically to address these issues and remove these barriers. It uses a 3-axis, piezo-driven nanopositioning stage to control sample motion and actively maintains the stability of the system using the image as the reference point for this stability.The broader impact/commercial potential of this project lies in making SR methods routinely useful to working biologists. These "game-changing" tools will advance our understanding of the molecular bases of disease pathologies, and enable far more exacting methods aimed at their treatments. The new insights will range from those in molecular virology and the development of safer and more effective vaccines, to the molecular mechanisms of neuronal signaling and learning and memory. In fact, it is hard to imagine an area of cell biology that will not be impacted by these emerging SR techniques. One of the pioneers of these methods has likened them to the Hubble telescope: they enable people to see things they simply could not see before. This analogy goes further: there is only one Hubble telescope, and currently very few SR-capable imaging systems, due to both the technical and economic barriers to their routine use. And while SR methods expose the physical limitations of microscopes in an acute manner, their stability and image acquisition requirements are not unique. Thus, this commercial system will be much more broadly useful: it will also enable focal-stability and molecular tracking at the nanometer-scale for any long-term imaging experiment.