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A. C. Ramsey, K. R. Seger, H. M. Proskin; Defining the Conjunctival Staining Method: Instillation Volume and Time Course to Assess Staining with Lissamine Green. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4681.
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A sufficient volume of lissamine green 1% is needed for the maximum conjunctival staining potential to be reached; however, excessive volumes of stain result in pooling in the conjunctival folds and overflow onto the cheek. The study objective was to determine the minimum volume of non-preserved lissamine green 1% and minimum time after instillation needed to achieve maximum staining potential of the conjunctiva.
A 30-subject, four-visit, crossover study to investigate the optimal volume of non-preserved lissamine green 1% and time needed to achieve maximum conjunctival staining was conducted. All subjects had 2.5 µL, 5 µL, and 10 µL of non-preserved lissamine green 1% instilled in both eyes during the course of the study and conjunctival staining was observed 1, 2, 3, 4, and 5 minutes after each instillation.
Significantly higher mean staining scores were observed for 5.0 µL when compared to 2.5 µL at 2 minutes (p=0.0195), 3 minutes (p=0.0004), 4 minutes (p=0.0074), and 5 minutes (p=0.0214). Significantly higher mean staining scores were observed for 10.0 µL when compared to 5.0 µL at 4 minutes (p=0.0268). A trend for a higher mean staining scores was also observed for 10.0 µL when compared to 5.0 µL at 5 minutes (p=0.0701). A comparison of adjacent times with respect to mean stain scores was made and a statistically significant difference was observed for all time points evaluated when comparing adjacent time points (p<0.0090). For each of the three volumes, the mean staining score peaked 2 minutes following instillation of lissamine green, followed by a reduction in mean staining scores at each minute thereafter.
These data suggest that 5.0 µL is the optimal volume of lissamine green 1% to assess conjunctival staining, and that conjunctival staining is best observed 2 minutes after instillation.
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