Hao Xiang, Allgeyer Edward S, Booth Martin J, Bewersdorf Joerg
Opt Lett. 2015 Aug 1;40(15):3627-30. doi: 10.1364/OL.40.003627.
IsoSTED nanoscopy, a variant of stimulated emission depletion (STED) microscopy, utilizes two opposing objective lenses and features the highest three-dimensional resolution of STED nanoscopes currently available. However, this technique is limited by axially repetitive side minima in the interference pattern of the depletion point-spread function (PSF), which can lead to ghost images. Here, we describe novel strategies to further improve the performance of isoSTED nanoscopy by reshaping the PSF. In particular, we propose employing moderate defocus on the depletion beam to reduce the side minima. Furthermore, we demonstrate a simplified alternative based on objective misalignment and quantitatively compare the expected performance between the two approaches.
等焦受激发射损耗（IsoSTED）纳米显微镜是受激发射损耗（STED）显微镜的一种变体，它使用两个相对的物镜，具有目前可用的STED纳米显微镜中最高的三维分辨率。然而，该技术受到耗尽点扩散函数（PSF）干涉图样中轴向重复的旁瓣最小值的限制，这可能导致重影。在这里，我们描述了通过重塑PSF来进一步提高IsoSTED纳米显微镜性能的新策略。特别是，我们建议对耗尽光束采用适度离焦以减少旁瓣最小值。此外，我们展示了一种基于物镜失调的简化替代方法，并定量比较了两种方法的预期性能。
