Quantitative three-dimensional confocal imaging of the cornea in situ and in vivo: System design and calibration

Research output: Contribution to journalArticle

70 Scopus citations

Abstract

A new depth encoding system (DES) is presented, which makes it possible to calculate, display, and record the z-axis position continuously during in vivo imaging using tandem scanning confocal microscopy (TSCM). In order to verify the accuracy of the DES for calculating the position of the focal plane in the cornea both in vitro and in vivo, we compared TSCM measurements of corneal thickness to measurements made using an ultrasonic pachymeter (UP, a standard clinical instrument) in both enucleated rabbit, cat, and human eyes (n = 15), and in human patients (n = 7). Very close agreement was found between the UP and TSCM measurements in enucleated eyes; the mean percent difference was 0.50 ± 2.58% (mean ± SD, not significant). A significant correlation (R=0.995, n=15, p<0.01) was found between UP and TSCM measurements. These results verify that the theoretical equation for calculating focal depth provided by the TSCM manufacturer is accurate for corneal imaging. Similarly, close agreement was found between the in vivo UP and TSCM measurements; the mean percent difference was 1.67 ± 1.38% (not significant), confirming that z-axis drift can be minimized with proper applanation of the objective. These results confirm the accuracy of the DES for imaging of the cornea both ex vivo and in vivo. This system should be of great utility for applications where quantitation of the three-dimensional location of cellular structures is needed.

Original languageEnglish (US)
Pages (from-to)45-49
Number of pages5
JournalScanning
Volume18
Issue number1
DOIs
StatePublished - Jan 1 1996

Keywords

  • Confocal microscopy
  • Cornea
  • In vivo imaging
  • Three-dimensional imaging

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Instrumentation

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