Multiplexing confocal technology

多重共焦技术

• 批准号: 486815-2015
• 负责人: Fang, Qiyin
• 金额: $ 4.37万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Idea to Innovation
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=690961
• 关键词: Multiplexing; confocal; technology

Comparing to conventional wide-field imaging microscopes, confocal microscopy hold significant advantages in image contrast enhancement, 3D sectioning capabilities, and compatibility with specialized detectors. Despite of being more costly, complicated to operate, and slow image acquisition rate, over the past decade, confocal fluorescence microscope has emerged as an important scientific research tool as well as applications in drug discovery, clinical pathology, etc. For applications such as live cell imaging, slow acquisition speed is a key barrier to adaption of confocal microscopy. While wide-field microscopy is typically faster, multiplexed confocal schemes such as using a spinning foci array can significantly increase the image acquisition rate. The moving foci array in the spinning disc, however, prevents the use of specialized discrete photo detectors arrays. We have developed a suite of technologies to generate, scan, and measure 100-400 confocal foci simultaneously, while is compatible with stationary discrete detectors. Another key feature of the technique is that it can be retrofitted to a conventional wide-field fluorescence microscope. The basic operation principle has been validated on a benchtop set up on live cell imaging. In this NSERC Idea-to-Innovation project, we plan to build a fully integrated, single-box prototype instrument that is fully compatible with conventional wide-field fluorescence microscopes from all four major manufacturers (Zeiss, Nikon, Olympus, and Leica). We will also explore subsequent commercialization pathways for such technology. In the next 12 months we plan to i) integration all modules into a single box, "manufacturing prototype", ii) characterize its performance on living cell imaging on different brand of commercial fluorescence microscopes and comparing them to commercial spinning disc confocal systems, and iii) evaluate different discrete photodetector array technologies and identify pathways to integration. Successful completion of the proposed prototype development will significantly strengthen the developed technology and its potential for the next phase commercialization plans.

与传统的广视场成像显微镜相比，共焦显微镜在图像对比度增强、3D切片能力以及与专用探测器的兼容性方面具有显著的优势。尽管共焦荧光显微镜成本较高、操作复杂、图像采集速度慢，但在过去的十年里，它已经成为一种重要的科学研究工具，并在药物开发、临床病理等方面得到了应用。对于活细胞成像等应用，采集速度慢是共焦显微镜应用的关键障碍。虽然广视场显微镜通常更快，但多路共焦方案，如使用旋转焦点阵列，可以显著提高图像采集速度。然而，旋转圆盘中的移动焦点阵列阻止了专用离散光电探测器阵列的使用。我们开发了一套技术，可以同时产生、扫描和测量100-400个共焦焦点，同时与固定的离散探测器兼容。这项技术的另一个关键特点是它可以改装成传统的广场荧光显微镜。基本工作原理已经在建立在活细胞成像的台式机上得到了验证。在NSERC的创意创新项目中，我们计划打造一个完全集成的单盒原型仪器，该仪器与所有四大制造商(蔡司、尼康、奥林巴斯和徕卡)的传统广场荧光显微镜完全兼容。我们还将探索此类技术随后的商业化途径。在接下来的12个月里，我们计划i)将所有模块集成到一个盒子中，“制造原型”，ii)表征其在不同品牌商业荧光显微镜上的活细胞成像性能，并将其与商业旋转圆盘共聚焦系统进行比较，以及iii)评估不同的分立光电探测器阵列技术，并确定集成的途径。成功完成拟议的原型开发将大大加强所开发的技术及其在下一阶段商业化计划中的潜力。