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.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.