Three-dimensional mapping of the orientation of collagen corneal lamellae in healthy and keratoconic human corneas using SHG microscopy

Year: 2017

Authors: Mercatelli R., Ratto F., Rossi F., Tatini F., Menabuoni L., Malandrini A., Nicoletti R., Pini R., Pavone FS., Cicchi R.

Autors Affiliation: National Institute of Optics, National Research Council (INO-CNR), Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy;
Institute of Applied Physics “N. Carrara” (IFAC-CNR), Via Madonna del Piano 10, 50019 Sesto Fiorentino, Italy; U.O. Oculistica Nuovo Ospedale S. Stefano, Via Suor Niccolina Infermiera 20, 59100 Prato, Italy; C.S.O S.r.l, Via degli Stagnacci 12/e, 50018 Scandicci, Italy; European Laboratory for Non-Linear Spectroscopy (LENS), Via Nello Carrara 1, 50019, Sesto Fiorentino, Italy; Department of Physics, University of Florence, Via G. Sansone 1, 50019 Sesto Fiorentino, Italy

Abstract: Keratoconus is an eye disorder that causes the cornea to take an abnormal conical shape, thus impairing its refractive functions and causing blindness. The late diagnosis of keratoconus is among the principal reasons for corneal surgical transplantation. This pathology is characterized by a reduced corneal stiffness in the region immediately below Bowman\’s membrane, probably due to a different lamellar organization, as suggested by previous studies. Here, the lamellar organization in this corneal region is characterized in three dimensions by means of second-harmonic generation (SHG) microscopy. In particular, a method based on a three-dimensional correlation analysis allows to probe the orientation of sutural lamellae close to the Bowman\’s membrane, finding statistical differences between healthy and keratoconic samples. This method is demonstrated also in combination with an epi-detection scheme, paving the way for a potential clinical ophthalmic application of SHG microscopy for the early diagnosis of keratoconus.


Volume: 10      Pages from: 75  to: 83

More Information: The research leading to these results has received funding from Tuscany Region and EU FP7 BiophotonicsPlus projects “LITE” (Laser Imaging of The Eye) and “LighTPatcH” (Led Technology in Photo Haemostasis), from the Italian Ministry for Education, University and Research in the framework of the Flagship Project NANOMAX, from the Italian Ministry of Health (GR-2011-02349626), from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement number 284464, from Fondazione Pisa and from Ente Cassa di Risparmio di Firenze.
KeyWords: Collagen; Diagnosis; Nonlinear optics; Transplantation (surgical), cornea; keratoconus; Lamellar organizations; Optical sectioning; Second harmonic generation microscopies (SHG); Statistical differences; Three dimensional mapping; Three-dimensional correlation, Harmonic generation, collagen, cornea; diagnostic imaging; human; keratoconus; microscopy; pathology; procedures; ultrastructure, Collagen; Cornea; Humans; Keratoconus; Microscopy
DOI: 10.1002/jbio.201600122

Citations: 46
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