title: The Effect of instilled fluorescein solution volume on the values and repeatability of TBUT measurements.
abstract: OBJECTIVE
  To investigate how the volume of instilled fluorescein solution alters invasive tear breakup time (TBUT) and the standard deviation (SD) of multiple recordings, and to determine the volume of fluid supplied by traditional fluorescein-impregnated strips (FS) and a micropipette calibrated to dispense 1 microL.
  
  
  METHODS
  TBUT was measured 3 times in the right eyes of 46 subjects following the instillation of 1, 2.7, and 7.4 microL of fluorescein solution by micropipette and the use of a wetted FS. The volume of fluid delivered to an eye by FS and a micropipette set for 1 microL was estimated by weighing each before and after use.
  
  
  RESULTS
  The volume of fluorescein solution instilled before measurement influenced TBUT (P < 0.001). Increasing the delivered volume from 1 to 2.7 microL lengthened TBUT (P < 0.001), but an additional increase in volume to 7.4 microL was not associated with a further change in TBUT (P = 0.50). Differences between TBUT values measured after using FS and a micropipette calibrated for 1 microL were not significant (P = 0.95). No significant differences were found in the repeatability of TBUT recordings between the instillation techniques used in this study (P = 0.18), although measurement variability increased with mean TBUT for all techniques. With the FS instillation technique described herein, the average volume of fluorescein solution they supplied was less than that delivered by a micropipette set for 1 microL (P = 0.017), but the range of volumes transferred by FS was greater.
  
  
  CONCLUSIONS
  Values of TBUT are dependent on the volume of fluorescein solution instilled before measurement; thus, attempts should be made to standardize this variable. Accurate estimation of TBUT requires more recordings to be averaged than are normally taken, unless breakup time is very short.
authors.affiliation: School of Optometry and Vision Sciences, Cardiff University, Cardiff, United Kingdom. JohnsonM2@Cardiff.ac.uk
authors.forename: Michael E
authors.initials: ME
authors.lastname: Johnson
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Murphy
mesh_terms: D000293:Adolescent; D000328:Adult; D003287:Contrast Media / Q000008:administration & dosage*; D003315:Cornea / Q000187:drug effects / Q000378:metabolism; D005260:Female; D019793:Fluorescein / Q000008:administration & dosage*; D006801:Humans; D008297:Male; D009883:Ophthalmic Solutions / Q000008:administration & dosage; D015203:Reproducibility of Results; D013666:Tears / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors
chemical_list: D003287:Contrast Media; D009883:Ophthalmic Solutions; D019793:Fluorescein