Three-dimensional nanoscopy of whole cells and tissues with in situ point spread function retrieval

Fan Xu, Donghan Ma, Kathryn P. MacPherson, Sheng Liu, Ye Bu, Yu Wang, Yu Tang, Cheng Bi, Tim Kwok, Alexander A. Chubykin, Peng Yin, Sarah Calve*, Gary E. Landreth, Fang Huang

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

64 Citations (Scopus)

Abstract

Single-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.

Original languageEnglish
Pages (from-to)531-540
Number of pages10
JournalNature Methods
Volume17
Issue number5
DOIs
Publication statusPublished - 1 May 2020
Externally publishedYes

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