Super-resolution imaging of flat-mounted whole mouse cornea

Zhen Cai, Yang Zhang, Zheyuan Zhang, Ki Hee Song, Lisa Beckmann, Ali Djalilian, Cheng Sun, Hao F. Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Super-resolution microscopy revolutionized biomedical research with significantly improved imaging resolution down to the molecular scale. To date, only limited studies reported multi-color super-resolution imaging of thin tissue slices mainly because of unavailable staining protocols and incompatible imaging techniques. Here, we show the first super-resolution imaging of flat-mounted whole mouse cornea using single-molecule localization microscopy (SMLM). We optimized immunofluorescence staining protocols for β-Tubulin, Vimentin, Peroxisome marker (PMP70), and Histone-H4 in whole mouse corneas. Using the optimized staining protocols, we imaged these four intracellular protein structures in the epithelium and endothelium layers of flat-mounted mouse corneas. We also achieved simultaneous two-color spectroscopic SMLM (sSMLM) imaging of β-Tubulin and Histone-H4 in corneal endothelial cells. The spatial localization precision of sSMLM in these studies was around 20-nm. This work sets the stage for investigating multiple intracellular alterations in corneal diseases at a nanoscopic resolution using whole corneal flat-mount beyond cell cultures.

Original languageEnglish (US)
Article number108499
JournalExperimental eye research
Volume205
DOIs
StatePublished - Apr 2021

Keywords

  • Corneal endothelium
  • Single-molecule localization microscopy
  • Super-resolution optical microscopy

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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