The American Medical Association's publication Journal of Ophthalmology published a letter to the editor from Dr. Alvis in 1985, in which he stated that Indiana University had received a research grant from the U.S. Air Force School of Aviation Medicine in 1959 to study the "oxygen saturation of retinal blood vessels by photographic methods."
Novotny brainstormed the concept of passing a dye through the system to establish the level of oxygen saturation and decided to use --- among other dyes --- sodium fluorescein. Alvis mixed a drop of his blood with fluorescein and shipped it to the Eli Lilly company for analysis. The results prompted the students to begin performing fluorescein studies on patients with diabetes and hypertension.
The first submission of their work to the American Journal of Ophthalmology in 1960 was rejected, but the two persevered. "They presented their technique for photographing fluorescence at the Association for Research in Ophthalmology on April 23, 1960, and an abstract appeared in the American Journal of Ophthalmology in July, 1960," writes The Ophthalmic Photographers' Society. The full description was published in 1961.