Yale University, Department of Biomedical Engineering, 55 Prospect St., New Haven, Connecticut 06511, USA.
J Biomed Opt. 2011 Oct;16(10):106009. doi: 10.1117/1.3641992.
Multiphoton microscopy of cleared tissue has previously been demonstrated to generate large three-dimensional (3D) volumetric image data on entire intact mouse organs using intrinsic tissue fluorescence. This technique holds great promise for performing 3D virtual biopsies, providing unique information on tissue morphology, and guidance for subsequent traditional slicing and staining. Here, we demonstrate the use of fluorescence lifetime imaging in cleared organs for achieving molecular contrast that can reveal morphologically distinct structures, even in the absence of knowledge of the underlying molecular source. In addition, we demonstrate the power of multimodal imaging, combining multiphoton fluorescence, second harmonic generation, and lifetime imaging to reveal exceptional morphological detail in an optically cleared mouse knee.
已证实，利用组织固有荧光对已被清除组织进行多光子显微镜检查，可以对整个完整的小鼠器官生成大型的三维（3D）容积图像数据。这项技术有望进行 3D 虚拟活组织检查，提供关于组织形态学的独特信息，并为后续的传统切片和染色提供指导。在这里，我们展示了在已被清除的组织中使用荧光寿命成像来实现分子对比，即使在不知道潜在分子来源的情况下，也可以揭示形态上不同的结构。此外，我们还展示了多模式成像的强大功能，将多光子荧光、二次谐波产生和寿命成像相结合，以揭示在光学清除的小鼠膝关节中非凡的形态细节。
