Moneron Gael, Hell Stefan W
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
Opt Express. 2009 Aug 17;17(17):14567-73. doi: 10.1364/oe.17.014567.
We report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with stimulated-emission depletion (STED). We demonstrate an easy-to-implement and promising laser combination based on a short-pulse laser source for two-photon excitation and a continuous-wave (CW) laser source for resolution enhancement. Images of fluorescent nanoparticles and the immunostained transcription regulator NF kappaB in mammalian cell nuclei exhibit resolutions of <50 nm and approximately 70 nm in the focal plane, respectively, corresponding to a 4-5.4-fold improvement over the diffraction barrier.
我们报告了通过将双光子激发（TPE）荧光显微镜技术与受激发射损耗（STED）技术相结合实现的亚衍射分辨率。我们展示了一种基于用于双光子激发的短脉冲激光源和用于分辨率增强的连续波（CW）激光源的易于实现且前景广阔的激光组合。荧光纳米颗粒的图像以及哺乳动物细胞核中免疫染色的转录调节因子NF-κB的图像在焦平面上的分辨率分别小于50 nm和约70 nm，比衍射极限提高了4至5.4倍。
