We report that high-density single-molecule super-resolution microscopy can be achieved with a conventional epifluorescence microscope set-up and a mercury arc lamp. The configuration termed as laser-free super-resolution microscopy (LFSM) is an extension of single-molecule localization microscopy (SMLM) techniques and allows single molecules to be switched on and off (a phenomenon termed as ‘blinking’), detected and localized. The use of a short burst of deep blue excitation (350–380 nm) can be further used to reactivate the blinking, once the blinking process has slowed or stopped. A resolution of 90 nm is achieved on test specimens (mouse and amphibian meiotic chromosomes). Finally, we demonstrate that stimulated emission depletion and LFSM can be performed on the same biological sample using a simple commercial mounting medium. It is hoped that this type of correlative imaging will provide a basis for a further enhanced resolution.

This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)’.

我们报告说，使用传统的落射荧光显微镜装置和汞弧灯可以实现高密度单分子超分辨率显微镜。称为无激光超分辨率显微镜 (LFSM) 的配置是单分子定位显微镜 (SMLM) 技术的扩展，允许打开和关闭单个分子（称为“闪烁”的现象）、检测和定位. 一旦眨眼过程减慢或停止，使用短脉冲深蓝色激发（350-380 nm）可进一步用于重新激活眨眼。在测试样本（小鼠和两栖动物减数分裂染色体）上实现了 90 nm 的分辨率。最后，我们证明可以使用简单的商业安装介质对相同的生物样品进行受激排放消耗和 LFSM。

本文是 Theo Murphy 会议问题“超分辨率结构照明显微镜（第 1 部分）”的一部分。