van de Linde Sebastian, Aufmkolk Sarah, Franke Christian, Holm Thorge, Klein Teresa, Löschberger Anna, Proppert Sven, Wolter Steve, Sauer Markus
Department of Biotechnology and Biophysics, Biozentrum, Julius-Maximilians-University Würzburg, Am Hubland, 97074 Würzburg, Germany.
Chem Biol. 2013 Jan 24;20(1):8-18. doi: 10.1016/j.chembiol.2012.11.004.
Super-resolution fluorescence imaging can provide insights into cellular structure and organization with a spatial resolution approaching virtually electron microscopy. Among all the different super-resolution methods single-molecule-based localization microscopy could play an exceptional role in the future because it can provide quantitative information, for example, the absolute number of biomolecules interacting in space and time. Here, small organic fluorophores are a decisive factor because they exhibit high fluorescence quantum yields and photostabilities, thus enabling their localization with nanometer precision. Besides past progress, problems with high-density and specific labeling, especially in living cells, and the lack of suited standards and long-term continuous imaging methods with minimal photodamage render the exploitation of the full potential of the method currently challenging.
超分辨率荧光成像能够以接近电子显微镜的空间分辨率洞察细胞结构和组织。在所有不同的超分辨率方法中,基于单分子的定位显微镜在未来可能发挥特殊作用,因为它可以提供定量信息,例如在空间和时间上相互作用的生物分子的绝对数量。在这里,小型有机荧光团是一个决定性因素,因为它们具有高荧光量子产率和光稳定性,从而能够以纳米精度对其进行定位。除了过去的进展,高密度和特异性标记方面的问题,尤其是在活细胞中,以及缺乏合适的标准和具有最小光损伤的长期连续成像方法,使得目前充分发挥该方法的全部潜力具有挑战性。