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Cellular in vivo 3D imaging of the cornea by confocal laser scanning microscopy. 2018
Sebastian Bohn, and
Karsten Sperlich, and
Stephan Allgeier, and
Andreas Bartschat, and
Ruby Prakasam, and
Klaus-Martin Reichert, and
Heinrich Stolz, and
Rudolf Guthoff, and
Ralf Mikut, and
Bernd Köhler, and
Oliver Stachs
Department of Ophthalmology, University Medical Center Rostock, 18057 Rostock, Germany.
We present an in vivo confocal laser scanning microscopy based method for large 3D reconstruction of the cornea on a cellular level with cropped volume sizes up to 266 x 286 x 396 µm3. The microscope objective used is equipped with a piezo actuator for automated, fast and precise closed-loop focal plane control. Furthermore, we present a novel concave surface contact cap, which significantly reduces eye movements by up to 87%, hence increasing the overlapping image area of the whole stack. This increases the cuboid volume of the generated 3D reconstruction significantly. The possibility to generate oblique sections using isotropic volume stacks opens the window to slit lamp microscopy on a cellular level.
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