Study of precorneal tear film thickness and structure by interferometry and confocal microscopy

Past measurements of precorneal tear film thickness in rabbits and humans gave values of 4-7 microns. These have been the accepted values. However, measurements presented in this report are much higher. Earlier techniques may have changed film thickness or not have fully included the layer containing mucus. Little is known about the mucus component. Thickness was measured in freshly killed animals of 10 species using two independent optical techniques. Coherent light was reflected from eyes and thickness was determined from spacing of interference fringes. Clear images of tissue structure were obtained with confocal microscopy, the tear film was visually identified, and thickness measured. Measurements by the two methods were closely correlated in each animal. No film was detected in trout or carp. In other species, thickness ranged from 9 microns in frogs to 15 microns in gerbils. Values are significantly larger than earlier estimates. Film thickness in three living rabbits was not significantly different from that measured shortly after death. There was no variation in thickness at five positions across the cornea in three species. Mucus content of rat tear film was examined by measuring thinning after application of 4, 8, and 20% (weight/volume) acetylcysteine, a mucolytic agent. Thickness was considerably reduced compared to controls, and the film almost completely removed by the highest concentration. The authors propose that the film is largely composed of mucus, not free aqueous solution. Our methods include such layers, and measurements are correspondingly larger than earlier ones.