Fluorescence nanoscopy uniquely combines minimally invasive optical access to the internal nanoscale structure and dynamics of cells and tissues with molecular detection specificity. While the basic physical principles of 'super-resolution' imaging were discovered in the 1990s, with initial experimental demonstrations following in 2000, the broad application of super-resolution imaging to address cell-biological questions has only more recently emerged. Nanoscopy approaches have begun to facilitate discoveries in cell biology and to add new knowledge. One current direction for method improvement is the ambition to quantitatively account for each molecule under investigation and assess true molecular colocalization patterns via multi-colour analyses. In pursuing this goal, the labelling of individual molecules to enable their visualization has emerged as a central challenge. Extending nanoscale imaging into (sliced) tissue and whole-animal contexts is a further goal. In this Review we describe the successes to date and discuss current obstacles and possibilities for further development.

荧光纳米技术独特地将对内部纳米尺度结构的微创光学访问以及细胞和组织的动力学与分子检测特异性完美地结合在一起。尽管“超分辨率”成像的基本物理原理是在1990年代发现的，随后在2000年进行了初步的实验演示，但直到最近才出现了将超分辨率成像用于解决细胞生物学问题的广泛应用。纳米技术已经开始促进细胞生物学的发现并增加新的知识。当前方法改进的方向是雄心勃勃地量化所研究的每个分子，并通过多色分析评估真正的分子共定位模式。为了实现这个目标，标记单个分子以使其可视化已成为一个主要挑战。将纳米级成像扩展到（切片的）组织和整个动物环境中是另一个目标。在这篇综述中，我们描述了迄今为止的成功，并讨论了当前的障碍和进一步发展的可能性。