Confocal imaging plate reader for advanced cell biology applications

具有成本效益的高通量成像是现代细胞生物学实验室的基石。然而，传统的荧光成像板阅读器缺乏在3D中解析复杂细胞细节的能力，而用于高通量筛选样品的共聚焦显微镜价格昂贵。本申请旨在资助Cytation C10旋转盘共聚焦成像板阅读器，以支持从微生物组到胚胎的高级细胞生物学倡议。Cytation C10是目前唯一可用的共聚焦成像平板阅读器，在一个紧凑的系统中结合了自动旋转盘共聚焦显微镜、宽视场显微镜和多模式检测。这种成像模式的组合将使我们的研究小组能够成像和量化复杂的细胞信息，从而更快地获得有意义的结果。申请人的nserc资助的研究侧重于细胞生物学的前沿方面。申请人面临着一个共同的挑战，即快速获得3D复杂细胞系统的高分辨率图像数据集，同时产生定量信息。研究主题包括细菌细胞形态和蛋白质定位的快速成像，化学遗传筛选的可视化输出，解剖参与细菌-宿主相互作用的信号传导和分子途径，以及研究内分泌和神经元发育。这种基础设施是迫切需要的，因为缺乏强大的、高通量的多模式成像，严重限制了他们的研究计划的范围。HQP的研究项目目前范围有限，并且由于需要在遥远的地点使用设备，或使用不一致且耗时的人工显微镜方法而受到阻碍。因此，快速准确地进行大规模筛查的能力是迫切需要的，并将使HQP能够提出更雄心勃勃的研究问题。申请人团队和他们的HQP代表了广泛的背景，反映了当今加拿大社会的多样性。

Cost-effective high throughput imaging is the cornerstone of modern cell biology laboratories. However, traditional fluorescence imaging plate readers lack the ability to resolve intricate cellular details in 3D, while confocal microscopes that function to screen samples in high throughput are prohibitively expensive. This application seeks to fund a Cytation C10 Spinning Disk Confocal Imaging Plate Reader to support advanced cell biology initiatives from microbiomes to embryos. The Cytation C10 is currently the only confocal imaging plate reader available and combines automated spinning disk confocal microscopy, widefield microscopy and multi-mode detection in one compact system. This combination of imaging modalities will enable our research groups to image and quantify complex cellular information to obtain meaningful results faster. The NSERC-funded research of the applicants focuses on cutting-edge aspects of cell biology. The applicants share a common challenge of obtaining high-resolution image datasets of complex cellular systems in 3D, rapidly while yielding quantitative information. The research themes include rapid imaging of morphology and protein localization in bacterial cells, visualizing outputs of chemical genetic screens, dissecting signalling and molecular pathways involved in bacterial-host interactions, and studying endocrine and neuronal development. This infrastructure is urgently needed as the lack of access to robust, high throughput multi-mode imaging has severely limited the scope of their research programs. The research projects of HQP are currently limited in scope and hampered by the need to access equipment at distant sites, or the use of inconsistent and time-consuming manual microscopy methods. The ability to perform large-scale screens rapidly and accurately is thus critically needed and will enable HQP to ask more ambitious research questions. The team of applicants and their HQP represent a range of backgrounds and reflect the diversity of Canadian society today.