We present the modular design and characterization of a multi-modality video-rate two-photon excitation (TPE) microscope based on integrating a digital micromirror device (DMD), which functions as an ultrafast beam shaper and random-access scanner, with a pair of galvanometric scanners. The TPE microscope system realizes a suite of new imaging functionalities, including (1) multi-layer imaging with 3D programmable imaging planes, (2) DMD-based wavefront correction, and (3) multi-focus optical stimulation (up to 22.7 kHz) with simultaneous TPE imaging, all in real-time. We also report the detailed optomechanical design and software development that achieves high level system automation. To verify the performance of different microscope functions, we have devised and performed imaging experiments on Drosophila brain, mouse kidney and human stem cells. The results not only show improved imaging resolution and depths via the DMD-based adaptive optics, but also demonstrate fast multi-focus stimulation for the first time. With the new imaging capabilities, e.g., tools for optogenetics, the multi-modality TPE microscope may play a critical role in the applications pertinent to neuroscience and biophotonics.

中文翻译：

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基于多模态DMD的双光子显微镜系统的设计

我们基于集成了数字微镜设备（DMD）的多模态视频速率双光子激发（TPE）显微镜的模块化设计和特性，该设备用作一对超快光束整形器和随机访问扫描仪检流扫描仪。TPE显微镜系统实现了一套新的成像功能，包括（1）具有3D可编程成像平面的多层成像，（2）基于DMD的波前校正和（3）多焦点光学刺激（最高22.7 kHz）同时进行TPE实时成像。我们还报告了实现高水平系统自动化的详细光机设计和软件开发。为了验证不同显微镜功能的性能，我们设计并在果蝇大脑上进行了成像实验，小鼠肾脏和人类干细胞。结果不仅显示出通过基于DMD的自适应光学器件改善的成像分辨率和深度，而且首次展示了快速的多焦点刺激。借助新的成像功能，例如光遗传学工具，多模态TPE显微镜可能在与神经科学和生物光子学相关的应用中发挥关键作用。