Washington University in St. Louis, Optical Imaging Laboratory, Department of Biomedical Engineering, One Brookings Drive, St. Louis, Missouri 63130.
Washington University School of Medicine, Department of Ophthalmology and Visual Sciences, Box 8096, St. Louis, Missouri 63110.
J Biomed Opt. 2014 Mar;19(3):36002. doi: 10.1117/1.JBO.19.3.036002.
The invention of green fluorescent protein and other molecular fluorescent probes has promoted applications of confocal and two-photon fluorescence microscopy in biology and medicine. However, exogenous fluorescence contrast agents may affect cellular structure and function, and fluorescence microscopy cannot image nonfluorescent chromophores. We overcome this limitation by integrating optical-resolution photoacoustic microscopy into a modern Olympus IX81 confocal, two-photon, fluorescence microscope setup to provide complementary, label-free, optical absorption contrast. Automatically coregistered images can be generated from the same sample. Imaging applications in ophthalmology, developmental biology, and plant science are demonstrated. For the first time, in a familiar microscopic fluorescence imaging setting, this trimodality microscope provides a platform for future biological and medical discoveries.
绿色荧光蛋白和其他分子荧光探针的发明推动了共聚焦和双光子荧光显微镜在生物学和医学中的应用。然而，外源性荧光对比剂可能会影响细胞结构和功能，并且荧光显微镜无法对非荧光生色团进行成像。我们通过将光分辨率光声显微镜集成到现代奥林巴斯 IX81 共聚焦、双光子、荧光显微镜设置中，以提供互补的、无标记的、光学吸收对比来克服这一限制。可以从同一样品生成自动配准的图像。展示了在眼科学、发育生物学和植物科学中的成像应用。这是第一次，在熟悉的显微镜荧光成像环境中，这种三模态显微镜为未来的生物学和医学发现提供了一个平台。
