Purpose Study the effect on the tear film in blepharospasm (BEB), facial hemispasm (FH), or aberrant regeneration (AR) treated with Botulinum Toxin (BTX-A). Methods A prospective study was used to evaluate the tear film in patients with BEB, FH, or AR treated with BTX-A. Schirmer tests, break-up time (BUT), optical coherence tomography (OCT) meniscus measurement, the Ocular Surface Disease Index (OSDI) questionnaire, and Oxford scale were documented before; 1 month after; and 3 months after BTX-A treatment. Comparisons were made with the Friedman test and Wilcoxon matched-pairs signed rank test was used. A p-value <0.05 was considered statistically significant. Results A total of 35 eyes from 27 patients were included. The mean patient age was 66.81 ± 12.94 years and 18 (66.7%) were female. Ten (37%) patients had BEB, six (22.2%) had FH, and 11 (40.74%) had AR. BTX-A improved the lid spasms. One month after BTX-A, Schirmer tests showed slight increments (Schirmer 1 p = 0.009; Schirmer 2 p = 0.05) and at 3 months they became similar to pre-treatment ( p = 0.5). The BUT test was not significantly different at 1 month ( p = 0.450) or at 3 months. On OCT 1 month after BTX-A, there was an increase in tear meniscus area ( p = 0.004), height ( p = 0.007), and depth ( p = 0.004), and at 3 months the measurements also became similar to the pre-BTX-A values. No significant changes in the OSDI ( p = 0.717) and Oxford scale ( p = 0.255). Conclusion OCT is a good tool to detect the increase in tear meniscus after periocular BTX-A in BEB, FH, and AR.