Goblet cell density estimate differences in impression cytology samples varies with different magnification of images.

PURPOSE: To assess the impact of using different microscope magnifications for the goblet cell density (GCD) estimates from conjunctival impression cytology (CIC) samples from healthy individuals

METHODS

In a prospective study, CIC specimens were collected from the superior bulbar conjunctiva (12 o'clock, 5mm from limbus) of 20 adult subjects (average age 22 years) onto Millicell-CM membranes and Giemsa stained. A region from each CIC filter containing reasonably high numbers of goblet cells was imaged by light microscopy at a final magnification of 400X and then the same region assessed at 200X and then 100X. The images were enlarged, the goblet cells marked and counted and GCD values/sq mm calculated.

RESULTS

The mean GCD estimates at 400X magnification, 200X and 100X were 644±180, 405±72 and 365±81 cells/sq mm respectively, and these values were statistically different (p<0.001).

CONCLUSIONS

As a result of non-uniform distribution, a strategy to select a 400X high power microscope field (HPF) that appears to include a moderate number of goblet cells will have a probability (by at least 20:1) that the GCD estimates will likely be higher compared to those at 200X or 100X, and the probability for higher GCD values is at least 15:1 comparing assessments made at 200X to 100X. Investigators should use only one magnification, with that of a medium power field (200X final magnification) likely being the most useful.