Detecting and locating a carrion resource is critical for the reproduction of necrophagous insects and initiating forensically important timelines. Blow flies (Diptera: Calliphoridae) primarily use olfactory cues in the form of volatile organic compounds to locate a suitable resource. Factors governing detecting and locating a resource have been studied using various behavior assays with modifications to suit the experiment design, such as the dual-choice cube olfactometer, which was examined in the current study. Systems optimization ensures biologically relevant and consistent results across replicates. In this study, two responses were measured: 1) leaving cube for either control or treatment and 2) choice between control and treatment. Phenotype (e.g., male, non-gravid, gravid) and total blow fly, Cochliomyia macellaria (Fabricius) (Diptera: Calliphoridae), number to respond were measured. Four parameters were assessed for their impact on response: 1) adjustment time in cube before trial, 2) trial length, 3) sugar/water presence, and 4) screening type in arms. Approximately, 70% of all phenotypes responded to liver with the 30-min adjustment period where only 50% responded with other adjustment periods.Trial length had a significant impact on response (35% increase in the 8 h trial compared to shorter durations); however, significant response to treatment was lost by increasing trial length.The presence of sugar/water decreased gravid and non-gravid response by 35% but did not impact males. Screening had no influence on overall or treatment response. Data indicate experiment design impacts fly response. Future studies should optimize parameters for their given fly population prior to initiating experiments.