Medda Rebecca, Jakobs Stefan, Hell Stefan W, Bewersdorf Jörg
Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, 37070 Göttingen, Germany.
J Struct Biol. 2006 Dec;156(3):517-23. doi: 10.1016/j.jsb.2006.08.013. Epub 2006 Sep 7.
The most prominent restrictions of fluorescence microscopy are the limited resolution and the finite signal. Established conventional, confocal, and multiphoton microscopes resolve at best approximately 200nm in the focal plane and only 500nm in depth. Additionally, organic fluorophores and fluorescent proteins are bleached after 10(4)-10(5) excitation cycles. To overcome these restrictions, we synergistically combine the 3- to 7-fold improved axial resolution of 4Pi microscopy with the greatly enhanced photostability of semiconductor quantum dots. Co-localization studies of immunolabeled microtubules and mitochondria demonstrate the feasibility of this approach for routine biological measurements. In particular, we visualize the three-dimensional entanglement of the two networks with unprecedented detail.
荧光显微镜最突出的限制是分辨率有限和信号有限。现有的传统显微镜、共聚焦显微镜和多光子显微镜在焦平面上的最佳分辨率约为200nm，在深度上仅为500nm。此外，有机荧光团和荧光蛋白在经历10⁴ - 10⁵次激发循环后会发生光漂白。为了克服这些限制，我们将4Pi显微镜轴向分辨率提高3至7倍的优势与半导体量子点大大增强的光稳定性进行了协同结合。对免疫标记的微管和线粒体进行的共定位研究证明了这种方法用于常规生物学测量的可行性。特别是，我们以前所未有的细节可视化了这两个网络的三维缠结。
