High resolution macroscopy (HRMac) of the eye using nonlinear optical imaging

Moritz Winkler; Bryan E. Jester; Chyong Nien-Shy; Dongyul Chai; Donald J. Brown; James V. Jester

doi:10.1117/12.847897

25 February 2010 High resolution macroscopy (HRMac) of the eye using nonlinear optical imaging

Moritz Winkler, Bryan E. Jester, Chyong Nien-Shy, Dongyul Chai, Donald J. Brown, James V. Jester

Proceedings Volume 7589, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X; 758906 (2010) https://doi.org/10.1117/12.847897
Event: SPIE LASE, 2010, San Francisco, California, United States

Abstract

Non-linear optical (NLO) imaging using femtosecond lasers provides a non-invasive means of imaging the structural organization of the eye through the generation of second harmonic signals (SHG). While NLO imaging is able to detect collagen, the small field of view (FoV) limits the ability to study how collagen is structurally organized throughout the larger tissue. To address this issue we have used computed tomography on optical and mechanical sectioned tissue to greatly expand the FoV and provide high resolution macroscopic (HRMac) images that cover the entire tissue (cornea and optic nerve head). Whole, fixed cornea (13 mm diameter) or optic nerve (3 mm diameter) were excised and either 1) embedded in agar and sectioned using a vibratome (200-300 um), or 2) embedded in LR White plastic resin and serially sectioned (2 um). Vibratome and plastic sections were then imaged using a Zeiss LSM 510 Meta and Chameleon femtosecond laser to generate NLO signals and assemble large macroscopic 3-dimensional tomographs with high resolution that varied in size from 9 to 90 Meg pixels per plane having a resolution of 0.88 um lateral and 2.0 um axial. 3-D reconstructions allowed for regional measurements within the cornea and optic nerve to quantify collagen content, orientation and organization over the entire tissue. We conclude that NLO based tomography to generate HRMac images provides a powerful new tool to assess collagen structural organization. Biomechanical testing combined with NLO tomography may provide new insights into the relationship between the extracellular matrix and tissue mechanics.

Citation Download Citation

Moritz Winkler, Bryan E. Jester, Chyong Nien-Shy, Dongyul Chai, Donald J. Brown, and James V. Jester "High resolution macroscopy (HRMac) of the eye using nonlinear optical imaging", Proc. SPIE 7589, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X, 758906 (25 February 2010); https://doi.org/10.1117/12.847897

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