Cotton fiber wax extraction yields of the contemporary accelerated solvent extraction (ASE) method were compared to the reference Soxhlet extraction method. Two Upland cotton varieties, referred to as the STV5458 B2F and the FM9180 B2F varieties grown in Lubbock, TX, USA, were examined. Effects of ASE extraction time and temperature were investigated. Characteristic environmental scanning electron microscopic (ESEM) images revealed fiber damage following both extraction methods, with the ASE method showing the least damage. A one-way ANOVA showed the two methods to be statistically different according to a 95% confidence level (p < 0.05) when combining analyses of the two cotton varieties.