Kaufmann Rainer, Müller Patrick, Hausmann Michael, Cremer Christoph
Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany.
Peptide-Chips and Nucleotide FISH, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany.
Micron. 2011 Jun;42(4):348-52. doi: 10.1016/j.micron.2010.03.006. Epub 2010 May 28.
Localisation microscopy methods allow to realize a light optical resolution far beyond the Abbe-Rayleigh limit of about 200 nm laterally and 600 nm axially. So far, this progress was achieved using labelling with appropriate fluorochromes and fluorescent proteins. Here, we describe for the first time that optical resolution of cellular structures in the λ/10 range (∼50 nm) can be achieved even in label-free cells. This was obtained using Spectral Precision Distance/Position Determination Microscopy (SPDM), a method based on the general principles of localisation microscopy. Besides a substantial resolution improvement of autofluorescent structures, SPDM revealed cellular objects which are not detectable under conventional fluorescence imaging conditions.
定位显微镜方法能够实现远超阿贝 - 瑞利极限的光学分辨率，横向约为200纳米，轴向约为600纳米。到目前为止，这一进展是通过使用合适的荧光染料和荧光蛋白标记来实现的。在此，我们首次描述，即使在无标记细胞中，也能够实现λ/10范围内（约50纳米）细胞结构的光学分辨率。这是通过光谱精密距离/位置测定显微镜（SPDM）实现的，该方法基于定位显微镜的一般原理。除了大幅提高自发荧光结构的分辨率外，SPDM还揭示了在传统荧光成像条件下无法检测到的细胞物体。
