Effects of Fluorescein Staining on Laser In Vivo Confocal Microscopy Images of the Cornea

Christine W Sindt,Jami R Kern,D Brice Critser,Trudy K Grout

doi:10.1155/2012/541974

Christine W Sindt, Jami R Kern + Show 2 more

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https://doi.org/10.1155/2012/541974

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Abstract

This study was designed to identify whether topical fluorescein, a common ophthalmic tool, affects laser in vivo confocal microscopy of the cornea, a tool with growing applications. Twenty-five eye care specialists were asked to identify presence or absence of fluorescein in 99 confocal micrographs of healthy corneas. Responses were statistically similar to guessing for the epithelium (48% ± 14% of respondents correct per image) and the subbasal nerve plexus (49% ± 11% correct), but results were less clear for the stroma. Dendritic immune cells were quantified in bilateral images from subjects who had been unilaterally stained with fluorescein. Density of dendritic immune cells was statistically similar between the unstained and contralateral stained eyes of 24 contact lens wearers (P = .72) and of 10 nonwearers (P = .53). Overall, the results indicated that fluorescein staining did not interfere with laser confocal microscopy of corneal epithelium, subbasal nerves, or dendritic immune cells.

Highlights

In vivo confocal microscopy of the cornea is making the transition from “bench to bedside” [1]
If confocal microscopy will be used in clinical studies or clinical practice, researchers need to know how confocal microscopy might interact with other ophthalmic measurements
Topical sodium fluorescein solution is often used in the evaluation of corneal integrity after contact lens wear [2], in the evaluation of corneal lesions associated with microbial keratitis [3], and in the evaluation of corneal integrity or tear film break-up time in patients with dry eye disease [4]

Summary

Introduction

In vivo confocal microscopy of the cornea is making the transition from “bench to bedside” [1]. If confocal microscopy will be used in clinical studies or clinical practice, researchers need to know how confocal microscopy might interact with other ophthalmic measurements. For ophthalmic sodium fluorescein in a solution with neutral pH, the peak excitation is at 490 nm and the peak emission is at 530 nm [5]. Laser in vivo confocal microscopy uses a wavelength of 670 nm [1]. While these 3 wavelengths appear to be well separated, absorption and emission spectra can have wide bands and secondary peaks. Some researches have investigated the effect of fluorescein on white-light, slitscanning in vivo confocal microscopy [6, 7], but no studies have yet reported the effect of fluorescein on laser-scanning in vivo confocal microscopy

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