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Assessment of the metabolism and morphology of the porcine cornea, lens and retina by 2‐photon imaging
Ist Teil von
Journal of biophotonics, 2018-07, Vol.11 (7), p.e201700324-n/a
Ort / Verlag
Weinheim: WILEY‐VCH Verlag GmbH & Co. KGaA
Erscheinungsjahr
2018
Link zum Volltext
Quelle
Wiley Online Library Journals Frontfile Complete
Beschreibungen/Notizen
Two‐photon imaging is a noninvasive imaging technique with increasing importance in the biological and medical fields since it allows intratissue cell imaging with high resolution. We demonstrate the feasibility of using a single 2‐photon instrument to evaluate the cornea, the crystalline lens and the retina based on their autofluorescence (AF). Image acquisition was performed using a custom‐built 2‐photon microscope for 5‐dimensional microscopy with a near infrared broadband sub‐15 femtosecond laser centered at 800 nanometers. Signals were detected using a spectral photomultiplier tube. The spectral ranges for the analysis of each tissue/layer AF were determined based on the spectra of each tissue as well as of pure endogenous fluorophores. The cornea, lens and retina are characterized at multiple depths with subcellular resolution based on their morphology and AF lifetime. Additionally, the AF lifetime of NAD(P)H was used to assess the metabolic activity of the cornea epithelium, endothelium and keratocytes. The feasibility to evaluate the metabolic activity of lens epithelial cells was also demonstrated, which may be used to further investigate the pathogenesis of cataracts. The results illustrate the potential of multimodal multiphoton imaging as a novel ophthalmologic technique as well as its potential as a diagnostic tool.
From left to right: 3‐dimensional representations of the porcine cornea, crystalline lens and the photoreceptors within the retina, based on individual and sequential images depicting volumes of approximately 95 × 95 ×120 μm3, 175 × 175 × 20 μm3, and 155 × 155 × 70 μm3, respectively.