title: Three-dimensional confocal microscopy and visualization of the in situ cornea.
abstract: The in situ cornea is an ideal test specimen to evaluate techniques for 3D reconstruction and visualization of unstained, unfixed, transparent living tissues from a stack of optical sections. The 0.4 mm thick transparent specimen has been optically sectioned into 365 sections using a confocal laser scanning microscope (CLSM) with a water immersion objective. Depth-dependent light attenuation due to absorption and scatter within the specimen was manually compensated at each sampled section. A water immersion microscope minimized the spherical aberrations that would have occurred with the use of an oil immersion objective. Isometric sampling resulted in near-cubic voxels, which compensated for the reduced microscopic resolution in the z axis as compared to x and y resolution.
authors.affiliation: Department of Anatomy and Cell Biology, Uniformed Services University of the Health Sciences, Bethesda, MD 20813.
authors.forename: B R
authors.initials: BR
authors.lastname: Masters
authors.forename: M A
authors.initials: MA
authors.lastname: Farmer
mesh_terms: D000818:Animals; D001082:Aqueous Humor / Q000166:cytology; D001485:Basement Membrane / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure; D003315:Cornea / Q000166:cytology / Q000648:ultrastructure*; D003319:Corneal Stroma / Q000648:ultrastructure; D004728:Endothelium, Corneal / Q000166:cytology; D004847:Epithelial Cells; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007834:Lasers; D008297:Male; D008854:Microscopy, Electron / Q000379:methods*; D011817:Rabbits; D012031:Refractometry; D014463:Ultrasonography