Chun Wanhee, Do Dukho, Gweon Dae-Gab
Department of Mechanical Engineering, KAIST, Daejeon 305-701, South Korea.
Rev Sci Instrum. 2013 Jan;84(1):013701. doi: 10.1063/1.4773232.
We developed a multimodal microscopy based on an optical scanning system in order to obtain diverse optical information of the same area of a sample. Multimodal imaging researches have mostly depended on a commercial microscope platform, easy to use but restrictive to extend imaging modalities. In this work, the beam scanning optics, especially including a relay lens, was customized to transfer broadband (400-1000 nm) lights to a sample without any optical error or loss. The customized scanning optics guarantees the best performances of imaging techniques utilizing the lights within the design wavelength. Confocal reflection, confocal fluorescence, and two-photon excitation fluorescence images were obtained, through respective implemented imaging channels, to demonstrate imaging feasibility for near-UV, visible, near-IR continuous light, and pulsed light in the scanning optics. The imaging performances for spatial resolution and image contrast were verified experimentally; the results were satisfactory in comparison with theoretical results. The advantages of customization, containing low cost, outstanding combining ability and diverse applications, will contribute to vitalize multimodal imaging researches.
我们开发了一种基于光学扫描系统的多模态显微镜，以便获取样品同一区域的多种光学信息。多模态成像研究大多依赖于商业显微镜平台，这种平台易于使用，但在扩展成像模式方面受到限制。在这项工作中，对光束扫描光学器件，特别是包括一个中继透镜进行了定制，以将宽带（400 - 1000纳米）光无光学误差或损失地传输到样品。定制的扫描光学器件保证了利用设计波长内的光的成像技术的最佳性能。通过各自实现的成像通道获得了共焦反射、共焦荧光和双光子激发荧光图像，以证明扫描光学器件中近紫外、可见光、近红外连续光和脉冲光的成像可行性。通过实验验证了空间分辨率和图像对比度的成像性能；与理论结果相比，结果令人满意。定制的优点，包括低成本、出色的组合能力和多样的应用，将有助于推动多模态成像研究的发展。
