Single-molecule manipulation and imaging platform

材料和试剂的宏观性质由分子尺度的相互作用决定。在某些领域，这些相互作用的细微细节可以完全改变化合物或材料的性质。例如，一种新的抗癌药物是否可以被设计成只与靶细胞表面受体结合，而不中断其他细胞过程？或者，可以改变有机纳米颗粒的吸水特性以增加其水分含量吗？分子间的相互作用可以从一个分子到下一个分子显著变化，使得平均值的测量具有欺骗性和无信息性。我们已经开发了一种新技术，命名为CLiC的“凸透镜诱导约束”，使简单和廉价的单分子成像能力的显微镜。虽然单分子成像技术已经存在（TIRF显微镜，光学镊子），但它们很笨重，并且它们使用的“分子系链”会使它们试图研究的反应产生偏差。CLiC要简单得多：分子被机械挤压到蚀刻在载玻片上的微小玻璃“盒子”中。它们可以无限期地保持在显微镜的视野中，从而能够进行深入，个别和无偏见的研究。加拿大初创公司ScopeSys正在将CLiC商业化，并已售出5套系统。为了使CLiC得到广泛的商业应用，我们将：1）完善CLiC的温度控制和试剂交换功能，以便在更广泛的条件下进行实验; 2）开发直观的分析软件，以便外部用户可以从他们的观察中提取有意义的单分子信息; 3）提高我们的“流动池”的可扩展性; 4）验证CLiC在几个重要应用中的使用。这项工作的成功将使加拿大研究人员能够尽早和容易地获得单分子成像方法，以加速他们的研究，即使在资金有限的实验室。此外，这项工作将使ScopeSys成长为一家加拿大公司，并为年轻的科学和技术专业人员创造高质量的就业机会。

Macroscopic properties of materials and reagents are governed by molecular-scale interactions. In some fields, subtle details of these interactions can completely change the property of a compound or material. For example, can a new cancer drug be engineered to bind only to the target cell surface receptor, without interrupting other cellular processes? Or, can the water-absorbing properties of an organic nanoparticle be altered to increase its moisture content? Molecular interactions can vary significantly from one molecule to the next, making measurements of averages deceptive and uninformative. We have developed a new technology, named CLiC for "convex lens-induced confinement", to bring simple and inexpensive single-molecule imaging capabilities to microscopy. While single-molecule imaging technologies already exist (TIRF microscopy, optical tweezers), they are cumbersome and their use of 'molecular tethers' biases the reactions they attempt to study. CLiC is much simpler: molecules are mechanically squeezed into tiny glass 'boxes' etched into a glass slide. They can be held indefinitely in the field of view of a microscope, enabling intensive, individual, and unbiased study. ScopeSys, a Canadian startup, is commercializing CLiC and has already sold 5 systems. To prepare CLiC for widespread commercial adoption we will 1) refine CLiC's temperature control and reagent exchange functionality to enable experimentation over a wider range of conditions, 2) develop intuitive analysis software so external users can extract meaningful single-molecule information from their observations, 3) improve the scalability of our 'flow cell', and 4) validate the use of CLiC in several important applications. Success of this work will allow a broad range of Canadian researchers early and easy access to single molecule imaging methods to accelerate their research, even in laboratories with limited funds. In addition, the work will allow ScopeSys to grow as a Canadian company and create high quality jobs for young science and technology professionals.