Fluorometric setup and method for studying the functional activity of corneal endotheliocytes

Abstract

Subject of study. Experimental methods for investigating the state of preservation and functionality of the corneal endothelium, which is determined by the water permeability of the cells and activity of ion pumps, mainly by sodium–potassium adenosine triphosphatase, were considered. Aim of study. The aim of this study was to identify significant measurable parameters such as the fluorescence intensity integrated over the entire cornea preparation and the temporal dynamic of this fluorescence. Additionally, a specialized experimental fluorometric setup was designed based on fiber optics. Method. Modifications were introduced to the composition and design of a universal experimental setup for the measurement of cell functions that enabled implementation of a fluorescence-based method for determination of the intracellular sodium concentration without processing the digital image of the cornea preparation. Main results. The specialized experimental setup and method were developed for investigating functions of the corneal endothelial cells and the intracellular signaling mechanisms whose activation or disfunction under conditions of a traumatic inflammation and after a hypothermic preservation can be related to the development of a corneal edema and a corneal graft disease. Practical significance. The specialized experimental setup and methods for functional testing of the corneal endothelial cells developed in this study can be applied to determine the degree of suitability of the cornea as a transplantation material. They can also be used as a system of tests for determining the effectiveness of the cornea preservation methods.