X-ray and scanning electron microscopic analysis of the structural composition of tear ferns

Dried tears from keratoconjunctivitis sicca eyes fail to exhibit the fern-like crystallization patterns observed with tears from eyes with normal tear function. To test our hypothesis that the extent of ferning depends on the ratio of salts to protein and mucin in the tear sample, dried tears from six normal subjects were subjected to scanning electron microscopy (SEM) and X-ray analyses. X-ray diffraction identified sodium chloride and potassium chloride as the major components of tear fern crystals. X-ray fluorescence detected the elements potassium, chlorine, calcium, and sulfur in the dried tear samples, with sulfur indicating the presence of protein and/or mucin. As well as confirming the presence of cubic fern nuclei, SEM revealed two kinds of material, having crystalline and globular appearances, that are hypothesized to be composed of salts and protein/mucin, respectively. Globular material appeared to block extension of crystal fern arms or to coat crystalline material, but did not crystallize. These findings suggest that tear fern crystals are composed of sodium and potassium chloride, with proteinaceous material controlling crystallization indirectly by coating crystal faces and blocking fern extension. This structural composition is consistent with the hypothesis that the ratio of salt to macromolecular species is an important determinant of tear ferning.