Laser-induced fluorescence whole column detection for capillary electrophoresis of biological species

我们已经致力于开发激光诱导荧光(LIF)全柱成像检测(WCID)生物物种毛细管电泳仪已有10年之久。最近，我们取得了重大突破，成功地开发了液芯波导-LIF-WCID毛细管等电聚焦(CIEF)系统。与UV-WCID相比，灵敏度提高了5个数量级。该系统已成功应用于蛋白质、抗体、病毒和细胞的分离和检测。然而，用于LCW-LIF-WCID设置的光学系统主要基于先前用于紫外线吸收检测的光学元件。十几年前的产品UV感光CCD相机，远远达不到我们团队继续开发这项技术所需的灵敏度和响应时间。需要提高检测灵敏度和图像采样率。此外，由于缺乏深紫外激光，限制了天然荧光在WCID中的应用，这是一种直接实现LIF检测的方法，避免了耗时和低成本的标记过程。因此，我们迫切需要现代的CCD摄像机、紫外线激光器、必要的光学元件和资金，以便在我们的设施中组装整个柱子成像系统。该系统将由至少两名研究生和一名博士后研究员使用，以维持研究势头，从而为CE，特别是CIEF开发高灵敏度的LIF-WCID技术，以监测和表征生物系统中发生的过程的动力学，如涉及蛋白质的反应和相互作用。我们还计划使用相同的组件构建基于表面增强拉曼散射(SERS)的新型全柱成像系统。我们建议构建的成像技术和系统是独特而强大的研究工具，这使加拿大在蛋白质组研究领域具有竞争力优势。总部位于安大略省的Convergent Bioscience公司承诺开发基于WCID系统的产品。使用拟议的LCW-LIF-WCID设备开发具有科学和商业价值的尖端应用程序将鼓励Convergent Bioscience在加拿大将这项技术商业化。

We have been working now for 10 years to develop laser-induced fluorescence (LIF) whole column imaging detection (WCID) equipment for capillary electrophoresis of biological species. Recently, we have made an important breakthrough, with a successful development of the capillary isoelectric focusing (CIEF) system with liquid-core waveguide (LCW)-LIF-WCID. The sensitivity was enhanced by up to 5 orders of magnitude as compared with UV-WCID. This system has been successfully applied to separation and detection of proteins, antibodies, viruses and cells. However, the optical system used for the LCW-LIF-WCID setup is mainly based on the optical components previously used for UV yabsorption detection. The UV sensitive CCD camera, a product of a dozen years ago, is far from the sensitivity and response time that our group requires to continue development of this technology. There is need for improvement of the detection sensitivity and image-sampling rate. In addition, the lack of a deep UV laser limits the utilization in WCID of native fluorescence, which is a straightforward method to achieve LIF detection avoiding the time-consuming and cost-inefficient labeling procedure. Therefore, we have a critical need for modern CCD camera, UV laser, necessary optical components and funds to put together the whole column imaging system in our facility. The system will be used by at least two graduate students and one post doctoral fellow to sustain momentum in research leading to the development of high-sensitivity LIF-WCID technique for CE, especially CIEF, of biological species present at trace levels, to monitoring and characterization of the dynamics of processes occurring in biological systems such as reactions and interactions involving proteins. We are also planning to use the same components to construct a new type of the whole column imaging system based on Surface-Enhanced Raman Scattering (SERS).  The imaging techniques and the systems we propose to construct are unique and powerful research tools, which gives Canada competitiveness edge in the field of proteomic research. There is commitment from Convergent Bioscience, Ontario based company to develop products based on the WCID systems.  Developments of cutting edge applications of scientific and commercial value using the proposed LCW-LIF-WCID equipment will encourage Convergent Bioscience to commercialize this technology in Canada.