Introduction: Different materials and technique exist to record the functional and physiologic border of the denture. There is a need to determine which among the various materials available would accurately record the borders width and height as compared to tissue conditioner. Aim: To compare and evaluate the morphology of the denture borders produced by three different commercially available border molding materials. Materials and Methods: This in-vivo study was conducted at KAHER’s KLE VK Institute of Dental Science, Belagavi, Karnataka, India, from November 2012 to June 2013. The study included 20 patients with previous dentures. The labial flange of 20 patients with the existing maxillary denture was trimmed 2 mm short of the sulcus and border molding was done with tissue conditioner. The patients were instructed to perform routine activity with the denture and asked to report after 1 day. Three custom trays were fabricated without spacer, 2 mm short of the labial border. Border molding was carried out with low fusing compound, putty addition silicone and pattern resin. The trays were beaded at 2 mm from the depth of the vestibule and were poured with die stone. Casts were obtained and with the standardised method, the cast were sectioned into six sections. Border morphology of the sulcus was viewed under stereomicroscope and under image analyser. The statistical analysis carried out was using Statistical Package for the Social Sciences (SPSS) software version 19.0 Inc. Chicago, IL, USA. The level of significance was 95% confidence with p-value <0.05 was considered. Analysis of Variance (ANOVA) test and Bonferroni multiple posts-hoc test was used to analyse the data. Results: The descriptive analysis showed that the mean area of tissue conditioner (111590.95 µm2 ) was least, next being pattern resin (131253.30 µm2 ) followed by Low fusing impression compound (154854.20 µm2 ) and addition silicone (190968.55 µm2 ) (p-value<0.001). Conclusion: The result of the study showed that tissue conditioner was the best material that could be used for border molding followed by pattern resin and low fusing compound.