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P<P, published online ahead of print November 5, 2009
(Investigative Ophthalmology and Visual Science. )
© 2009 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.09-4186
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Automated Assessment of Keratocyte Density in Stromal Images from the ConfoScan 4 Confocal Microscope

Jay W McLaren,1 William M Bourne,2 and Sanjay V Patel3

1Dept. of Ophthalmology Research, Mayo Clinic, Rochester, United States 2Ophthalmology, Mayo Clinic, Rochester, United States 3Ophthalmology, Mayo Clinic, Rochester, United States

Correspondence: Jay McLaren, Email: mclaren.jay{at}mayo.edu

Abstract

Purpose: We developed a program to determine cell densities in images from the ConfoScan 4 (Nidek, Inc., Freemont, CA) confocal microscope and compared densities to those determined from images from the Tandem Scanning Confocal Microscope (Tandem Scanning Corporation, Reston, VA).

Methods: A program was developed that used image processing routines to identify stromal cell nuclei in images from the ConfoScan 4 confocal microscope. Cell selection parameters were set to match cell densities from the program with those determined manually in 15 normal corneas of 15 volunteers. The program was tested on scans from 16 other normal volunteers and 17 volunteers 3 years after LASIK. Cell densities were compared to densities determined by manual assessment and from scans by a Tandem Scanning confocal microscope in the same corneas.

Results: The difference in cell density between the automatic and manual assessment was -539 ± 3,005 cells/mm3 (mean ± SD, p=0.11) in the 16 test corneas. Densities estimated from the ConfoScan 4 agreed with those from the Tandem Scanning confocal microscope in all regions of the stroma except in the anterior 10%, where the ConfoScan 4 indicated a 30% lower density.

Conclusions: Differences in anterior stromal cell density between the ConfoScan 4 and the Tandem Scanning confocal microscopes can be explained by the different optical designs. The lower spatial resolution of the ConfoScan 4 limits its ability to resolve thin layers. The adaptation of our earlier cell-counting program to the ConfoScan 4 provides a timesaving, objective, and reproducible means of determining stromal cell densities in images from the ConfoScan 4.

Key Words: keratocytes • corneal stroma • confocal microscopy • cell density






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