Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada.
Biophys J. 2009 Dec 16;97(12):3224-8. doi: 10.1016/j.bpj.2009.09.038.
We report applications of two-photon excitation fluorescence (2PEF) microscopy with subdiffraction-limit resolution for green-fluorescent-protein-tagged cell imaging. The microscope integrates 2PEF microscopy and stimulated emission depletion microscopy in one microscope that has the benefits of both techniques: intrinsic three-dimensional resolution, confined photobleaching, and subdiffraction-limit resolution. The subdiffraction-limit resolution was demonstrated by resolving green-fluorescent-protein-tagged caveolar vesicles located within a distance shorter than the diffraction limit of a regular 2PEF microscope, which is approximately 250 nm even with the best optics. The full width at half-maximum of the effective point-spread function for the 2PEF microscope was estimated to be approximately 54 nm.
我们报告了双光子激发荧光（2PEF）显微镜在具有亚衍射极限分辨率的绿色荧光蛋白标记细胞成像中的应用。该显微镜将 2PEF 显微镜和受激发射损耗显微镜集成在一台显微镜中，这两种技术都具有各自的优点：固有三维分辨率、受限光漂白和亚衍射极限分辨率。通过分辨位于距离小于常规 2PEF 显微镜的衍射极限的距离内的绿色荧光蛋白标记的小窝囊泡，证明了亚衍射极限分辨率，即使使用最佳光学器件，该距离也约为 250nm。2PEF 显微镜的有效点扩散函数的半最大值全宽估计约为 54nm。
