title: In situ osmometry: validation and effect of sample collection technique.
abstract: OBJECTIVE
  To compare tear film osmolarity measurements between in situ and vapor pressure osmometers. Repeatability of in situ measurements and the effect of sample collection techniques on tear film osmolarity were also evaluated.
  
  
  METHODS
  Osmolarity was measured in one randomly determined eye of 52 healthy participants using the in situ (TearLab Corporation, San Diego, CA) and the vapor pressure (Vapro 5520; Wescor, Inc., Logan, UT) osmometers. In a subset of 20 participants, tear osmolarity was measured twice on-eye with the in situ osmometer and was additionally determined on a sample of nonstimulated collected tears (3 μL) with both instruments.
  
  
  RESULTS
  Mean (SD) tear film osmolarity with the in situ osmometer was 299.2 (10.3) mOsmol/L compared with 298.4 (10) mmol/kg with the vapor pressure osmometer, which correlated moderately (r = 0.5, P < 0.05). Limits of agreement between the two instruments were -19.7 to +20.5 mOsmol/L. Using collected tears, measurements with the vapor pressure osmometer were marginally higher (mean [SD], 303.0 [11.0] vs 299.3 [8.0] mOsmol/L; P > 0.05) but correlated well with those using the in situ osmometer (r = 0.9, P < 0.05). The mean (SD) osmolarity of on-eye tears was 5.0 (6.6) mOsmol/L higher than that of collected tears, when both measurements were conducted with the in situ osmometer. This was a consistent effect because the measurements correlated well (r = 0.65, P < 0.05).The in situ osmometer showed good repeatability with a coefficient of repeatability of 9.4 mOsmol/L (r = 0.8, P < 0.05).
  
  
  CONCLUSIONS
  Correlation between the two instruments was better when compared on collected tear samples. Tear film osmolarity measurement is influenced by the sample collection technique with the osmolarity of on-eye tears being higher than that of collected tears. This highlights the importance of measuring tear film osmolarity directly on-eye. The in situ osmometer has good repeatability for conducting this measurement.
authors.affiliation: School of Optometry and Vision Science, The University of New South Wales, Sydney, New South Wales, Australia. m.gokhale@unsw.edu.au
authors.forename: Moneisha
authors.initials: M
authors.lastname: Gokhale
authors.forename: Ulrike
authors.initials: U
authors.lastname: Stahl
authors.forename: Isabelle
authors.initials: I
authors.lastname: Jalbert
mesh_terms: D000328:Adult; D015352:Dry Eye Syndromes / Q000378:metabolism / Q000503:physiopathology; D004867:Equipment Design; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009994:Osmolar Concentration; D061286:Osmometry / Q000379:methods*; D012016:Reference Values; D015203:Reproducibility of Results; D013048:Specimen Handling; D013666:Tears / Q000737:chemistry*; D055548:Vapor Pressure; D055815:Young Adult