Chapter 4: Super-resolution Fluorescence Imaging

Our current understanding of living systems has advanced to the level of individual cells. However, there is still a pressing need to more precisely visualize the microstructure of cells, as well as the dynamic actions of biomolecules therein, including molecular translocation and chemical modifications (e.g. epigenetics and post-translational modification) at the molecular level in real time. Fluorescence microscopy permits non-invasive imaging of biomolecules in living cells, but its resolution is limited by diffraction. While electron microscopy provides a higher resolution, it requires fixed cells to be used, thus compromising the fidelity of the structural and functional information of biomacromolecules. Super-resolution fluorescence microscopy (SRFM) has emerged as a groundbreaking technology that addresses the diffraction limit issue, and enables high-resolution imaging of subcellular as well as biomolecular structures. The advent of SRFM has significantly advanced our understanding of how cellular structures are exquisitely organized, and how biomolecules interact with each other to modulate cell fate. This chapter will discuss the recent progress in the use of various SRFM-based techniques for the structural and functional imaging of cells. The diverse fluorescent agents developed for SRFMs will also be summarized.