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. 2020 May 26;11(6):3263-3273.
doi: 10.1364/BOE.391815. eCollection 2020 Jun 1.

In vivo corneal and lenticular microscopy with asymmetric fundus retroillumination

Timothy D Weber  1 , Jerome Mertz  1   2
Affiliations

In vivo corneal and lenticular microscopy with asymmetric fundus retroillumination

Timothy D Weber et al. Biomed Opt Express. .

Abstract

We describe a new technique for non-contact in vivo corneal and lenticular microscopy. It is based on fundus retro-reflection and back-illumination of the crystalline lens and cornea. To enhance phase-gradient contrast, we apply asymmetric illumination by illuminating one side of the fundus. The technique produces micron-scale lateral resolution images across a 1 mm diagonal field of view in the central cornea. We show representative images of the epithelium, the subbasal nerve plexus, large stromal nerves, dendritic immune cells, endothelial nuclei, and the anterior crystalline lens, demonstrating the potential of this instrument for clinical applications.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
System optical diagram. PBS: Polarizing beamsplitter.
Fig. 2.
Fig. 2.
Illumination beam, shown in red, at the pupil/cornea (left) and at the fundus (right). Diffusely reflected light from the fundus retroilluminates the central cornea including the microscope field of view (FOV).
Fig. 3.
Fig. 3.
Subbasal nerve plexus of a 28-year-old male in the central cornea. (A) Full FOV (820 x 580 µm) and (B) indicated area in detail. Arrows indicate suspected dendritic cells. See Visualization 1 for a laterally-stabilized video excerpt. (C) A separate area seen later in the image sequence showing complex anastamoses. Scale bars: (A) 100 µm, (B) & (C) 25 µm.
Fig. 4.
Fig. 4.
Other epithelial structures. (A) The tear layer of a 26-year-old female. (B) Superficial and (C) basal epithelial cells in a 28-year-old male. (D) Dendritic immune cells in a 57-year-old male. Scale bars: 25µm.
Fig. 5.
Fig. 5.
Stromal nerves in a 28-year-old male. (A) Stromal nerve segment just anterior to view shown in (B). Arrows indicate the same branch point. (C) Stromal nerve in focus near the center and out of focus at the top and bottom. Scale bars: 50 µm.
Fig. 6.
Fig. 6.
Endothelium of a 26-year-old female. Bright/dark circles are nuclei anterior or posterior to the focal plane (see Visualization 2 for laterally-stabilized through-focus). Scale bar: 100 µm.
Fig. 7.
Fig. 7.
Crystalline lens (A) epithelium and (B) anterior fibers just below the epithelium. Scale bars 50 µm.
See this image and copyright information in PMC

References

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