Gulka, Sebastian

Project title: Confocal laser scanning microscope imaging of the ex vivo, intact, unfixed mouse cornea

Department: Biology

Faculty supervisor: Dr. Bob Chow

"Previous work in the lab involved the development of a novel method for analyzing the cornea utilizing a lipophilic fluorescent membrane dye, SGC5, paired with confocal laser microscopy. This method uses unfixed enucleated eyes of mice which were recently sacrificed, in order to preserve (as best as possible) the anatomical structure of the cornea. Approaches which include fixation of the eye may result in by-products-such as corneal bubbling or distortion-which can lead to inaccurate corneal quantification. Current highlights of this technique include visualization of corneal thickness and curvature, and robust corneal nerve imaging. Imaging corneal nerves presents the possibility of corneal nerve quantification as a function of this method. These metrics are all useful in contrasting the differences between normal and disease states which affect the cornea. My current research is aimed at refining this technique using mouse models by trialing approaches which alter SGC5 concentration, corneal application strategy for SGC5, and other variables. All corneal nerves are seemingly not labelled with this approach; therefore, I also seek to determine what fraction of corneal nerves that this technique labels."