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Surface topography and 3-dimensional confocal profiling of femtosecond and mechanical microkeratome dissected posterior human corneal buttons

Session Details

Session Title: Corneal Femtosecond

Session Date/Time: Monday 07/10/2013 | 14:30-16:30

Paper Time: 15:06

Venue: Elicium 1 (First Floor)

First Author: : M.Dickman THE NETHERLANDS

Co Author(s): :    M. van Maris   F. van Marion   F. van den Biggelaar   T. Berendschot   R. Nuijts  

Abstract Details


To determine the feasibility of confocal profiling in measuring surface roughness and obtaining three-dimensional surface reconstructions of posterior lamellar grafts for endothelial keratoplasty and to measure and compare the surface roughness of posterior corneal buttons harvested by a femtosecond (FS) laser platform and two mechanical microkeratomes (MK).


University Eye Clinic Maastricht, Maastricht, The Netherlands, Eindhoven University of Technology, Eindhoven, The Netherlands, Euro Cornea Bank, Beverwijk, The Netherlands.


Endothelial buttons were harvested from 18 human corneas unsuitable for transplantation with the Intralase FS60 laser (Abbott Medical Optics, Santa Anna, Ca.; n=4) and the Moria ALTK (Moria, Antony, France; n=6) or the Gebauer SLc MKs (Gebauer Medizintechnik, Neuhausen, Germany; n=8). Following dissection the endothelial buttons were fixed in 3.0% glutaraldehyde and examined using a confocal profiler (Sensofar PLu 2300, Terrassa, Spain) and environmental scanning electron microscopy (ESEM) (FEI-Quanta 600; Hillsboro, OR). Linear mixed model analysis was used to quantify the difference in surface roughness between the different dissection techniques


Confocal profiling was able to obtain quantitative surface roughness measurements and high resolution 3-dimensional reconstructions of the central 2x1.5 mm2 area of the dissected buttons. ESEM evaluation allowed wide field analysis at lower magnification which corresponded well with the 3-dimensional confocal reconstructions. Surface roughness of FS dissected buttons (RMS =1.89 µm) was significantly higher compared with the Moria (RMS=1.05 µm) and Gebauer MKs (RMS=0.92 µm) (p<0.01). However, there was no significant difference between the surface roughness of the MK dissected buttons (p=0.29) despite ultrastructural differences between the buttons as demonstrated by the 3-dimensional reconstruction and ESEM analysis.


FS laser dissection of corneal buttons results in significantly rougher stromal surfaces compared to mechanical MKs. MK dissected corneal buttons did not differ in surface roughness. Confocal profiling, a novel non-contact optical profiling method, enables submicron surface roughness measurements and 3-dimensional reconstructions of large areas of human corneal buttons, corresponding well with qualitative 2-dimensional ESEM imaging. This innovative and objective technology may be useful for quality judgement of lamellar dissection techniques.

Financial Interest:


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