Share Email Print
cover

Proceedings Paper

High resolution macroscopy (HRMac) of the eye using nonlinear optical imaging
Author(s): Moritz Winkler; Bryan E. Jester; Chyong Nien-Shy; Dongyul Chai; Donald J. Brown; James V. Jester
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper 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.

Paper Details

Date Published: 25 February 2010
PDF: 7 pages
Proc. SPIE 7589, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X, 758906 (25 February 2010); doi: 10.1117/12.847897
Show Author Affiliations
Moritz Winkler, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)
Bryan E. Jester, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)
Chyong Nien-Shy, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)
Dongyul Chai, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)
Donald J. Brown, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)
James V. Jester, The Gavin Herbert Eye Institute, Univ. of California Irvine Medical Ctr. (United States)


Published in SPIE Proceedings Vol. 7589:
Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X
Alexander Heisterkamp; Joseph Neev; Stefan Nolte; Rick P. Trebino, Editor(s)

© SPIE. Terms of Use
Back to Top