Ultramicroscopy: development and outlook
Ultramicroscopy: development and outlook
复制标题
DOI：
10.1117/1.nph.2.4.041407
复制
发表时间：
2015-10-01
期刊：
影响因子：
5.3
通讯作者：
Niendorf, Axel
中科院分区：
文献类型：
--
作者：
Dodt, Hans-Ulrich;Saghafi, Saiedeh;Niendorf, Axel
我们提出了一个概述，我们开发的超显微镜技术。从100年前的发展开始，我们设计了一个光片显微镜和一个化学清洗来成像完整的小鼠大脑。利用我们的3DISCO透明化技术可以保存小鼠脑中绿色荧光蛋白（GFP）标记神经元的荧光，并获得高分辨率的三维（3-D）记录。超显微镜也被用来成像整个小鼠胚胎和苍蝇。我们改进了我们的光片显微镜的光学切片，产生更长和更薄的光片与非球面光学。为了获得高分辨率的图像，我们校正了可用的空气显微镜物镜，用于高折射率的清洁溶液。讨论了如何应用受激发射耗尽技术最终实现片层显微镜的超分辨。还讨论了通过记录表达cfos-GFP的小鼠脑来成像脑功能。最后，我们展示了人类乳腺癌的光片显微镜的第一个3-D记录，作为医学诊断的应用。(C)作者。
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. (C) The Authors.