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. 2020 Sep 18;11(10):5689-5700.
doi: 10.1364/BOE.397615. eCollection 2020 Oct 1.

Multiwavelength confocal laser scanning microscopy of the cornea

Sebastian Bohn  1   2   3   4   5 , Karsten Sperlich  1   3   4   6 , Thomas Stahnke  1   3 , Melanie Schünemann  1   3 , Heinrich Stolz  2 , Rudolf F Guthoff  1   3 , Oliver Stachs  1   3
Affiliations

Multiwavelength confocal laser scanning microscopy of the cornea

Sebastian Bohn et al. Biomed Opt Express. .

Abstract

Confocal reflectance microscopy has demonstrated the ability to produce in vivo images of corneal tissue with sufficient cellular resolution to diagnose a broad range of corneal conditions. To investigate the spectral behavior of corneal reflectance imaging, a modified laser ophthalmoscope was used. Imaging was performed in vivo on a human cornea as well as ex vivo on porcine and lamb corneae. Various corneal layers were imaged at the wavelengths 488 nm, 518 nm, and 815 nm and compared regarding image quality and differences in the depicted structures. Besides the wavelength- and depth-dependent scattering background, which impairs the image quality, a varying spectral reflectance of certain structures could be observed. Based on the obtained results, this paper emphasizes the importance of choosing the appropriate light source for corneal imaging. For the examination of the epithelial layers and the endothelium, shorter wavelengths should be preferred. In the remaining layers, longer wavelength light has the advantage of less scattering loss and a potentially higher subject compliance.

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Conflict of interest statement

The authors declare that there are no conflicts of interest related to this article.

Figures

Fig. 1.
Fig. 1.
A: The RCM 2.0 in conjunction with the SPECTRALIS platform (dashed line separates both components). B: Simplified schematic of a confocal SLO with three laser diodes operating at different wavelengths (LD – laser diode, D – detector).
Fig. 2.
Fig. 2.
Comparison of the HRT + RCM 2.0 and SPECTRALIS + RCM 2.0 combinations showing superficial cells of a human cornea.
Fig. 3.
Fig. 3.
Comparison of in vivo human cornea images of superficial epithelial cells acquired at three distinct wavelengths as indicated. The histograms of the subimages in the yellow dashed squares were adjusted to the full intensity range for a consistent comparison of the image quality.
Fig. 4.
Fig. 4.
Comparison of in vivo human cornea images of epithelium acquired at three distinct wavelengths as indicated. The histograms of the subimages in the yellow dashed squares were adjusted to the full intensity range for a consistent comparison of the image quality.
Fig. 5.
Fig. 5.
Comparison of in vivo human cornea images of the subbasal nerve plexus acquired with three distinct wavelengths as indicated. The histograms of the subimages in the yellow dashed squares were adjusted to the full intensity range for a consistent comparison of the image quality.
Fig. 6.
Fig. 6.
Comparison of in vivo human cornea images of anterior stroma acquired at three distinct wavelengths as indicated. The histograms of the subimages in the yellow dashed squares were adjusted to the full intensity range for a consistent comparison of the image quality.
Fig. 7.
Fig. 7.
Comparison of in vivo human cornea images of endothelium acquired at three distinct wavelengths as indicated. The histograms of the subimages in the yellow dashed squares were adjusted to the full intensity range for a consistent comparison of the image quality.
Fig. 8.
Fig. 8.
Comparison of in vivo human cornea images of endothelium captured with specular and confocal microscopy at three different wavelengths. The histogram of each image is stretched for better comparison.
Fig. 9.
Fig. 9.
In vivo (human) and ex vivo (porcine, lamb) images of superficial epithelial cells acquired at blue and near-infrared wavelengths.
Fig. 10.
Fig. 10.
In vivo (human) and ex vivo (porcine, lamb) images of t stroma acquired at blue and near-infrared wavelengths.
Fig. 11.
Fig. 11.
In vivo (human) and ex vivo (porcine, lamb) images of endothelium acquired at blue and near-infrared wavelengths.
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References

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