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To evaluate measurement reliability of the meibomian gland (MG) tissue area during infrared video meibography (IR-VM) of the lower lid.This is a retrospective analysis, using ImageJ, of IR-VM of 17 lids of 11 consecutive patients (age 56.6 ± 15.9 years; 9 females and 2 males) focusing on the presence of a statistically significant variation of the MG tissue area and the MG tissue area relative to the total measurement field (TMF) secondary to inadvertent lid distortion and/or an altered vertical gaze direction during a single-image acquisition video sequence.For lids with distortions, 10 of 10 lids with distortion showed a statistically significant difference (P < 0.0001–0.0136) in the mean individual gland area (MIGA, % change: 16.2%–114.1%) and/or the ratio of MIGA to TMF (% change: 19.8%–108.5%) during its video sequence. For the altered vertical gaze direction, 7 of 9 lids individually showed a statistically significantly increased MIGA/TMF with the globe in primary gaze compared with upgaze (P = 0.001–0.0156), whereas collectively as a group, all 9 lids showed a significant increase in MIGA/TMF (P < 0.0001, % change: 1.1%–38.9%). Two lids from the same patient were evaluated for both types of distortion.IR-VM of lower lids revealed statistically significant unreliability of measuring the MG tissue area from inadvertent lid distortion or an altered vertical globe gaze direction during meibography. These data suggest that, lacking context seen when using the video, still-shot nonvideo infrared meibography images of the lower lid, and related data, may be specious and should be used with caution in drawing conclusions of the change in the MG area over time, to avoid misguided clinical decision making.