Multiphoton (MP) 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) from collagen and two photon excited fluorescence (TPEF) from elastin. To map the collagen and elastin organization within the eye we have used computed tomography on optical and mechanical sectioned tissue to generate high-resolution macroscopic (HRMac) images that cover the entire tissue. 3-Dimensional tomographs varied in size from 9 to 90 Meg pixels per plane, had a pixel resolution of 0.44 µm lateral and 2.0 µm axial, and covered regions extending from 9 mm2 to 100 mm2. These reconstructions allowed for regional measurements within the cornea and optic nerve to quantify collagen and elastin content, orientation and organization over the entire tissue that could be directly correlated to tissue stiffness using biomechanical testing.

© 2011 Optical Society of America

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