Gray Institute for Radiation Oncology & Biology, Dept. Of Oncology, University of Oxford, Oxford, UK.
J Microsc. 2013 Aug;251(2):154-67. doi: 10.1111/jmi.12057. Epub 2013 Jun 12.
We describe a microscopy design methodology and details of microscopes built to this 'open' design approach. These demonstrate the first implementation of time-domain fluorescence microscopy in a flexible automated platform with the ability to ease the transition of this and other advanced microscopy techniques from development to use in routine biology applications. This approach allows easy expansion and modification of the platform capabilities, as it moves away from the use of a commercial, monolithic, microscope body to small, commercial off-the-shelf and custom made modular components. Drawings and diagrams of our microscopes have been made available under an open license for noncommercial use at http://users.ox.ac.uk/~atdgroup. Several automated high-content fluorescence microscope implementations have been constructed with this design framework and optimized for specific applications with multiwell plates and tissue microarrays. In particular, three platforms incorporate time-domain FLIM via time-correlated single photon counting in an automated fashion. We also present data from experiments performed on these platforms highlighting their automated wide-field and laser scanning capabilities designed for high-content microscopy. Devices using these designs also form radiation-beam 'end-stations' at Oxford and Surrey Universities, showing the versatility and extendibility of this approach.
我们描述了一种显微镜设计方法和根据该“开放式”设计方法制造的显微镜的详细信息。这些显微镜展示了时域荧光显微镜在灵活自动化平台中的首次实现，该平台具有将这种技术和其他先进显微镜技术从开发阶段轻松过渡到常规生物学应用的能力。这种方法允许轻松扩展和修改平台功能，因为它不再使用商业的、单片的显微镜主体，而是使用小型的、商用现货的和定制的模块化组件。我们的显微镜的图纸和图表已根据开放许可证提供，可供非商业用途在 http://users.ox.ac.uk/~atdgroup 上使用。已经使用该设计框架构建了几个自动化高通量荧光显微镜实现，并针对多孔板和组织微阵列进行了优化。特别是，三个平台以自动化方式通过时间相关单光子计数实现了时域 FLIM。我们还展示了在这些平台上进行的实验数据，突出了它们为高通量显微镜设计的自动化宽场和激光扫描功能。在牛津大学和萨里大学，使用这些设计的设备也形成了辐射束“终端站”，展示了这种方法的多功能性和可扩展性。
