Post conductive keratoplasty visualization of rabbit cornea by multiphoton microscopy

Conductive keratoplasty (CK) is a new refractive surgery for presbyopia and hyperopia patients. By applying radio frequency current at the peripheral regions of cornea, collagen, the most abundant composition of corneal stroma, shrinks due to the heat generated. The shrinkage at the periphery alters the corneal architecture and achieves clearer focus for near vision. In this work we use multiphoton microscopy to observe the post surgery structure variation at both submicron resolution and over a large region within the tissue. Since collagen can be induced to generate strong second harmonic generation (SHG) signal, multiphoton excitation provide direct visualization of collagen orientation within corneal stroma. In addition, since the SHG intensity of collagen tissue deteriorates with increasing thermal damage [1-3], our methodology can be used to characterize the extent of corneal stroma damage from the CK procedure. Finally, the influence of CK on the morphology and distribution of keratocytes can also be investigated by detecting multiphoton excited autofluorescence from the cells.