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

Experimental eye research 2021 Apr

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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. Copyright © 2021 Elsevier Ltd. All rights reserved.

Citation

Zhen Cai, Yang Zhang, Zheyuan Zhang, Ki-Hee Song, Lisa Beckmann, Ali Djalilian, Cheng Sun, Hao F Zhang. Super-resolution imaging of flat-mounted whole mouse cornea. Experimental eye research. 2021 Apr;205:108499