Multiphoton autofluorescence and SHG microscopy have been demonstrated to be an effective technique for resolving, respectively, the cellular and collagen structures within the ocular surface of ex vivo porcine eyes. SHG imaging resolved the difference in structural orientations between corneal and sclera collagen fibers. Specifically, the corneal collagen is organized in a depth-dependent fashion, whereas the scleral collagen is randomly packed. Because this technique does not require histologic preparation procedures, it has the potential to be applied for in vivo studies with minimal disturbance to the eye.

Two-photon autofluorescence and second harmonic generation (SHG) microscopy are useful imaging techniques for studying tissue components. In this work, we apply this imaging modality to study porcine eye. In particular, we use SHG microscopy to investigate the structural changes to excised porcine corneas and found that this technique is useful for studying its structural changes under thermal treatment.

This purpose of this study is to demonstrate the feasibility of using multiphoton microscopy in imaging eye surface. Spec$cally, the cornea, limbus. conjunctiva, and sclera were imaged using multiphoton inducedfluorescence and second-harmonic generation (SHC) imaging.

This purpose of this study is to demonstrate the feasibility of using multiphoton microscopy in ophthalmologic imaging. Without the introduction of extrinsic fluorescence molecules, multiphoton induced autofluorescence and second harmonic generation signals can be used to obtain useful structural information of normal and diseased corneas. Our work can potentially lead to the in vivo application of multiphoton microscopy in investigating corneal physiology and pathologies.