Extending the spatiotemporal resolution of super-resolution microscopies using photomodulatable °uorescent proteins

In the past two decades,various super-resolution(SR)microscopy techniques have been developed to break the di®raction limit using subdi®raction excitation to spatially modulate the°uorescence emission.Photomodulatable°uorescent proteins(FPs)can be activated by light of speci¯c wavelengths to produce either stochastic or patterned subdi®raction excitation,resulting in improved optical resolution.In this review,we focus on the recently developed photomodulatable FPs or commonly used SR microscopies and discuss the concepts and strategies for optimizing and selecting the biochemical and photophysical properties of PMFPs to improve the spatiotemporal resolution of SR techniques,especially time-lapse live-cell SR techniques.

荧光蛋白与超分辨显微成像

超分辨显微成像技术使细胞生物学进入到了一个全新的时代,但如何进一步提高超分辨显微成像技术的时空分辨率仍是光学领域需要解决的重要问题。目前为止几乎所有的超分辨显微成像技术都依赖于荧光探针,光调控荧光蛋白作为一类特殊的荧光探针,可以被不同波长的激发光所激活,产生随机或者特殊结构样式的信号。利用这些信息,透镜系统的空间分辨率得到了提高。通过总结光调控荧光蛋白的各类参数,从荧光探针入手,寻找进一步提高成像系统空间分辨率的方法与策略,为选取适当的荧光探针提供建议,并且阐述了荧光蛋白与超分辨显微成像技术之间的关系。