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Abstract

Bioengineering

A Custom Multiphoton Microscopy Platform for Live Imaging of Mouse Cornea and Conjunctiva

Published: May 17th, 2020

DOI:

10.3791/60944

1Department of Biomedical Engineering, National Taiwan University, 2Institute of Pharmacology, College of Medicine, National Taiwan University, 3Molecular Imaging Center, National Taiwan University, 4Department of Dermatology, National Taiwan University Hospital, and College of Medicine, 5Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, 6Department of Ophthalmology, Chang Gung Memorial Hospital, 7College of Medicine, Chang Gung University

* These authors contributed equally

Abstract

Conventional histological analysis and cell culture systems are insufficient to simulate in vivo physiological and pathological dynamics completely. Multiphoton microscopy (MPM) has become one of the most popular imaging modalities for biomedical study at cellular levels in vivo, advantages include high resolution, deep tissue penetration and minimal phototoxicity. We have designed an MPM imaging platform with a customized mouse eye holder and a stereotaxic stage for imaging ocular surface in vivo. Dual fluorescent protein reporter mouse enables visualization of cell nuclei, cell membranes, nerve fibers, and capillaries within the ocular surface. In addition to multiphoton fluorescence signals, acquiring second harmonic generation (SHG) simultaneously allows for the characterization of collagenous stromal architecture. This platform can be employed for intravital imaging with accurate positioning across the entire ocular surface, including cornea and conjunctiva.

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Keywords Multiphoton Microscopy

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