The investigation of human tear film proteins and lipids is of value for the elucidation of contact lens incompatibilities, tear film instabilities, dry eye syndrome and various other eye diseases. Improved efficient methods for the investigation of human tear film proteins and lipids are presented in this paper. Tear proteins were examined by ultrathin sodium dodecyl sulfate-polyacrylamide gel electrophoresis, celluloacetate gel, isoelectric focusing, and high-performance liquid chromatography. The methods differ in sensitivity, resolution, convenience and reproducibility. Tear lipids were examined by high-performance thin-layer chromatography, and good separation into the major lipid classes was achieved. With this method it is possible to examine the lipids present in tears of an individual subject and not just in pools made up from the tears of several persons.