Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
Pathologie Hamburg-West , Lornsenstraße 4, 22767 Hamburg, Germany.
Neurophotonics. 2015 Oct;2(4):041407. doi: 10.1117/1.NPh.2.4.041407. Epub 2015 Nov 9.
We present an overview of the ultramicroscopy technique we developed. Starting from developments 100 years ago, we designed a light sheet microscope and a chemical clearing to image complete mouse brains. Fluorescence of green fluorescent protein (GFP)-labeled neurons in mouse brains could be preserved with our 3DISCO clearing and high-resolution three-dimensional (3-D) recordings were obtained. Ultramicroscopy was also used to image whole mouse embryos and flies. We improved the optical sectioning of our light sheet microscope by generating longer and thinner light sheets with aspheric optics. To obtain high-resolution images, we corrected available air microscope objectives for clearing solutions with high refractive index. We discuss how eventually super resolution could be realized in light sheet microscopy by applying stimulated emission depletion technology. Also the imaging of brain function by recording of mouse brains expressing cfos-GFP is discussed. Finally, we show the first 3-D recordings of human breast cancer with light sheet microscopy as application in medical diagnostics.
我们介绍了我们开发的超微镜技术的概述。从 100 年前的发展开始，我们设计了一种光片显微镜和一种化学透明化方法，以对完整的老鼠大脑进行成像。使用我们的 3DISCO 透明化方法可以保留绿色荧光蛋白（GFP）标记的神经元的荧光，并且可以获得高分辨率的三维（3-D）记录。超微镜还可用于对整个老鼠胚胎和苍蝇进行成像。我们通过使用非球面光学元件生成更长更薄的光片，来改进光片显微镜的光学切片。为了获得高分辨率的图像，我们针对具有高折射率的透明化溶液对现有的空气显微镜物镜进行了校正。我们讨论了如何通过应用受激发射损耗技术最终在光片显微镜中实现超分辨率。还讨论了通过记录表达 cfos-GFP 的老鼠大脑来记录大脑功能的成像。最后，我们展示了光片显微镜在医学诊断中的首例 3-D 记录的人类乳腺癌。
