Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo.

Abstract:

PURPOSE:To demonstrate the application of multiphoton fluorescence (MF) and second harmonic generation (SHG) microscopy for ex vivo characterization of the structural alterations of human corneas due to keratoconus. METHODS:Buttons of keratoconic corneas derived from penetrating keratoplasty were sent for structural analysis with a custom-built multiphoton microscope. Fluorescence detected within the cytoplasm and the SHG signal obtained from collagen were used to demonstrate the morphologic changes in the corneal specimens. RESULTS:The fluorescent epithelial cells around the apical area were elongated and were aligned parallel to the adjacent collagen fibers. Parallel and centripetal distribution patterns of stromal collagen bundles were demonstrated at different depths within the keratoconic corneas. CONCLUSIONS:MF and SHG microscopy provides three-dimensional structural analysis of keratoconus ex vivo. It may provide important morphologic information for the investigation of the pathogenesis of keratoconus and may have potential in a clinical setting as an in vivo diagnostic and monitoring system for advancing keratoconus.

authors

Tan HY,Sun Y,Lo W,Lin SJ,Hsiao CH,Chen YF,Huang SC,Lin WC,Jee SH,Yu HS,Dong CY

doi

10.1167/iovs.06-0386

subject

Has Abstract

pub_date

2006-12-01 00:00:00

pages

5251-9

issue

12

eissn

0146-0404

issn

1552-5783

pii

47/12/5251

journal_volume

47

pub_type

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