Leibniz Institute of Photonic Technology, Albert-Einstein-Str.9, 07745 Jena, Germany.
Methods Appl Fluoresc. 2016 Nov 22;4(4):045005. doi: 10.1088/2050-6120/4/4/045005.
Fluorescent microscopy methods linked to the reassignment principle as image scanning microscopy (ISM), re-scan confocal (RSC), optical photon reassignment (OPRA) and instant structured illumination microscopy (iSIM) have the potential to replace confocal microscopy as the standard microscopy technique. Photon reassignment methods are known to link the most important properties in biological imaging as resolution, sensitivity, imaging speed and combinability with fluorophores in an elegant way. On the example of OPRA, we show how this method could be easily extended to the third dimension. If OPRA is used in combination with a structured illumination pattern the sectioning ability can be improved while maintaining the very high signal intensity. We present a detailed analysis about the imaging properties of OPRA in three dimensions and show experimental results on biological samples.
荧光显微镜方法与重分配原理相关联，如图像扫描显微镜（ISM）、再扫描共聚焦（RSC）、光学光子重分配（OPRA）和即时结构照明显微镜（iSIM），具有取代共聚焦显微镜成为标准显微镜技术的潜力。光子重分配方法以一种优雅的方式将生物成像中最重要的特性，如分辨率、灵敏度、成像速度和与荧光团的组合性联系起来。以 OPRA 为例，我们展示了如何轻松地将该方法扩展到第三个维度。如果将 OPRA 与结构照明模式结合使用，可以在保持非常高的信号强度的同时提高切片能力。我们对 OPRA 在三维中的成像特性进行了详细分析，并展示了生物样本的实验结果。
