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Journal of Microscopy | 1992

Comparison of in vivo and ex vivo cellular structure in rabbit eyes detected by tandem scanning microscopy

J. V. Jester; W. M. Petroll; R. M. R. Garana; Michael A. Lemp; H. D. Cavanagh

Using the tandem scanning microscope, in vivo confocal microscopic images of living eyes were compared to images obtained from ex vivo, freshly enucleated or fixed tissue in the rabbit. In the normal cornea, microscopic details of the superficial epithelium, basal lamina, stromal fibrocyte nuclei, nerves and endothelial cell borders were easily discernible. Removal of the eye from the intact animal resulted in loss of detail with distortion of the normal structural interrelationships within the corneal stroma whilst enhancing details of the corneal epithelium. Formalin fixation further enhanced details of the basal and suprabasal corneal epithelial cell nuclei and the stromal fibrocyte cell borders whilst inducing prominent brightly reflecting folds in the thickened stroma with concomitant enhancement of the edge contrast of the collagen lamellae. These changes appeared to be related, in part, to hydration of the cornea and artefactual pooling of water between structures that may enhance reflectivity by increasing the difference between the refractive index of the cellular and extracellular elements. We conclude that microscopic examination of ex vivo preparations of corneal tissue, although providing increased resolution similar to conventional light microscopic techniques, significantly altered the normal structural relationships and could lead to erroneous measurements of the physiological properties of the tissue as compared to in vivo microscopy of undisturbed, intact tissue.


Cornea | 1995

Quantitative assessment of anteroposterior keratocyte density in the normal rabbit cornea

W. M. Petroll; Boettcher K; P A Barry; H. D. Cavanagh; James V. Jester

The anteroposterior keratocyte density distribution in the rabbit cornea was measured. Unsectioned tissue blocks from the central cornea of five rabbits were stained with propidium iodide and imaged using a Leica laser scanning confocal microscope. A z-series of images was acquired in each sample, from anterior to posterior stroma in either 3− or 8-μm steps. Software was developed to allow interactive marking of the keratocyte nuclei within each section of the z-series and for calculating cell density. For convenience, cell density was expressed as the number of cells per corneal volume element (CVE), where CVE is a newly defined volume unit with x, y, and z dimensions of 250, 250, and 10 μm, respectively. The calculated keratocyte density was 20.2 ± 1.0 cells/CVE (n = 5), which is equivalent to 32,360 ± 1,660 cells/mm3. The greatest density was underneath the epithelium (26.3 ± 2.5 cells/CVE), the density then decreased linearly with depth to 15.2 ± 1.4 cells/CVE; there was a slight increase in density pre-Descemets membrane to 18.5 ± 3.5 cells/CVE. A 30% decrease in cell density over the entire anteroposterior stromal thickness was observed. To facilitate statistical analysis, the cell density was averaged over 5% thickness intervals from anterior to posterior cornea. A significant difference in mean cell density of these intervals was found (ANOVA, n = 20, p < 0.01). To further assess the density distribution, linear regression analysis was performed. A significant correlation was found between keratocyte density and stromal depth (R = −0.94, n = 20, p < 0.05). We conclude that in the rabbit, keratocyte cell density is maximal underlying the epithelium and progressively decreases from anterior to posterior cornea.


Cornea | 1994

In vivo osmotic pertubation of intercellular fluid channels in the rabbit corneal endothelium

Peter M. Andrews; James V. Jester; W. M. Petroll; P A Barry; Hideji Ichijima; H. D. Cavanagh

An in vivo rabbit corneal model was used to evaluate morphological changes in the corneal endothelium associated with osmotically increasing fluid movement from the anterior chamber into the stroma. When the corneal stroma is rendered more hypertonic than normal by immersing the scraped epithelial side of the cornea in a hypertonic sucrose solution, intercellular channels and apical pores at the Y-junctions between endothelial cells become greatly enlarged. The foregoing changes are reversible and do not appear to result in damage to the corneal endothelium. These observations suggest that specific intercellular channels in the corneal endothelium may provide pathways for the movement of fluid from the aqueous humor into the stroma.


The CLAO journal : official publication of the Contact Lens Association of Ophthalmologists, Inc | 1990

Confocal microscopy of the living eye

H. D. Cavanagh; J. V. Jester; Essepian J; W. J. Shields; Michael A. Lemp


Journal of Electron Microscopy Technique | 1991

In vivo, real-time confocal imaging

J. V. Jester; Peter M. Andrews; W. M. Petroll; Lemp Ma; H. D. Cavanagh


Journal of Anatomy | 1995

Temporal, 3-dimensional, cellular anatomy of corneal wound tissue

James V. Jester; W. M. Petroll; P A Barry; H. D. Cavanagh


Scanning | 1994

In vivo confocal imaging: General principles and applications

W. M. Petroll; James V. Jester; H. D. Cavanagh


Experimental pathology | 1996

In vivo confocal imaging.

W. M. Petroll; James V. Jester; H. D. Cavanagh


The CLAO journal : official publication of the Contact Lens Association of Ophthalmologists, Inc | 1991

Continued contact lens wear in patients with giant papillary conjunctivitis

Lustine T; Bouchard Cs; H. D. Cavanagh


Scanning | 1991

In vivo 3-dimensional reconstruction of corneal fibroblasts using confocal microscopy

W. M. Petroll; J. Hill; H. D. Cavanagh; J. V. Jester

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Michael A. Lemp

Georgetown University Medical Center

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P A Barry

University of Texas Southwestern Medical Center

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Boettcher K

University of Texas Southwestern Medical Center

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