Xu, FanMa, DonghanMacPherson, Kathryn P.Liu, ShengBu, YeWang, YuTang, YuBi, ChengKwok, TimChubykin, Alexander A.Yin, PengCalve, SarahLandreth, Gary E.Huang, Fang2021-12-132021-12-132020-05Xu, F., Ma, D., MacPherson, K. P., Liu, S., Bu, Y., Wang, Y., Tang, Y., Bi, C., Kwok, T., Chubykin, A. A., Yin, P., Calve, S., Landreth, G. E., & Huang, F. (2020). Three-dimensional nanoscopy of whole cells and tissues with in situ point spread function retrieval. Nature Methods, 17(5), 531–540. https://doi.org/10.1038/s41592-020-0816-x1548-7105https://hdl.handle.net/1805/27165Single-molecule localization microscopy is a powerful tool for visualizing subcellular structures, interactions and protein functions in biological research. However, inhomogeneous refractive indices inside cells and tissues distort the fluorescent signal emitted from single-molecule probes, which rapidly degrades resolution with increasing depth. We propose a method that enables the construction of an in situ 3D response of single emitters directly from single-molecule blinking datasets, and therefore allows their locations to be pinpointed with precision that achieves the Cramér-Rao lower bound and uncompromised fidelity. We demonstrate this method, named in situ PSF retrieval (INSPR), across a range of cellular and tissue architectures, from mitochondrial networks and nuclear pores in mammalian cells to amyloid-β plaques and dendrites in brain tissues and elastic fibers in developing cartilage of mice. This advancement expands the routine applicability of super-resolution microscopy from selected cellular targets near coverslips to intra- and extracellular targets deep inside tissues.enPublisher PolicyAnimalsCartilageCell NucleusArticle