Spinning disk confocal microscope for live-cell imaging of pathogen-host cell interactions and biofilm development

我们的研究小组研究了高度动态的细胞和分子过程，作为细胞和病原体之间相互作用的一部分，在免疫反应和生物膜的形成过程中。这些过程发生的速度很快，但周期从几个小时到几天不等。为了推进我们的研究，我们迫切需要一种能够以高空间和时间分辨率捕捉这些过程的图像的显微镜。多伦多大学斯卡伯勒校区目前还没有具备这些特征的显微镜。为此，我们要求提供资金，将现有的徕卡DMI6000B倒置显微镜升级为能够满足我们当前需求和长期研究的共焦显微镜系统。拟议的升级包括：将安装在徕卡DMI6000B倒置显微镜上的X-Light V2旋转盘共焦扫描头，以及具有七条激光线路的发光二极管照明器。重要的是，升级将包括将显微镜封闭在Chamlide HX丙烯酸孵化器笼子中，该笼子将为病原体、毒素和细胞的活细胞实验提供适当的生物安全容器。它还将配备舞台上的环境控制，这对于长时间的活细胞成像至关重要，以确保细胞存活以及成像的焦点稳定性。所要求的系统将立即对我们的研究生产力产生积极影响，并将显著提高培训能力和高资质人员(HQP)的新型基础设施暴露。

Our research groups study highly dynamic cellular and molecular processes occurring as part of the interplay between cells and pathogens, during the immune response and in the formation of biofilms. These processes occur at high speeds, but over periods that can range from several hours or days. To advance our studies, we have an urgent need for a microscope that can capture images of these processes with high spatial and temporal resolution. A microscope of these characteristics is not currently available at the University of Toronto, Scarborough campus. For this reason, we request funding to upgrade a pre-existing Leica DMI6000B inverted microscope into a confocal microscopy system that can fulfill our immediate requirements and long-term research. The proposed upgrade includes: a X-Light V2 Spinning Disk Confocal scanner head to be installed on our Leica DMI6000B inverted microscope, as well as a Light Diode Illuminator with seven laser lines. Importantly, the upgrade will include enclosing the microscope in a Chamlide HX acrylic incubator cage that will provide the appropriate Biosafety containment for live cell experiments with pathogens, toxins and cells. This will also be equipped with on-stage environmental controls, which is crucial for live cell imaging over long periods to ensure cell viability, as well as focal stability for imaging. The requested system will have an immediate positive impact on our research productivity and will substantially improve the training capacity and novel infrastructure exposure for High Qualified Personnel (HQP).**************