Olivier Nicolas, Keller Debora, Rajan Vinoth Sundar, Gönczy Pierre, Manley Suliana
Laboratory for Experimental Biophysics, School of Basic Sciences, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
Biomed Opt Express. 2013 May 14;4(6):885-99. doi: 10.1364/BOE.4.000885. Print 2013 Jun 1.
3D STORM is one of the leading methods for super-resolution imaging, with resolution down to 10 nm in the lateral direction, and 30-50 nm in the axial direction. However, there is one important requirement to perform this type of imaging: making dye molecules blink. This usually relies on the utilization of complex buffers, containing different chemicals and sensitive enzymatic systems, limiting the reproducibility of the method. We report here that the commercial mounting medium Vectashield can be used for STORM of Alexa-647, and yields images comparable or superior to those obtained with more complex buffers, especially for 3D imaging. We expect that this advance will promote the versatile utilization of 3D STORM by removing one of its entry barriers, as well as provide a more reproducible way to compare optical setups and data processing algorithms.
3D STORM是超分辨率成像的领先方法之一，横向分辨率低至10纳米，轴向分辨率为30 - 50纳米。然而，进行此类成像有一个重要要求：使染料分子闪烁。这通常依赖于使用含有不同化学物质和敏感酶系统的复杂缓冲液，这限制了该方法的可重复性。我们在此报告，商业封片剂Vectashield可用于Alexa - 647的STORM成像，并且能产生与使用更复杂缓冲液获得的图像相当或更优的图像，特别是对于三维成像。我们预计这一进展将通过消除其进入障碍之一来促进3D STORM的广泛应用，并提供一种更具可重复性的方法来比较光学设置和数据处理算法。
