Little work has been done to quantify the number of eggs oviposited by blow flies (Diptera: Calliphoridae) in studies examining colonization behavior. Egg counting methods currently available are time-consuming and destructive. This study used ImageJ software and analysis of covariance to relate the volume of egg masses to the number of eggs laid by three different blow fly species: Lucilia sericata (Meigen), Phormia regina (Meigen), and Chrysomya rufifacies (Macquart). Egg mass volume, species, and the interaction of species and egg mass volume all affected the number of blow fly eggs deposited in egg masses. Both species identity and egg mass volume are important when predicting egg number, as such a single regression equation cannot be used to estimate egg number for these three species. Therefore, simple linear regression equations were determined for each species. The volume of individual eggs was incorporated into the model, yet differences between species were observed, suggesting that the orientation of the eggs oviposited by multiple conspecific females within egg masses influences egg estimates. Based on our results, we expect that imaging software can be used for other blow fly species, as well as other insect species; however, equations specific to each species must be developed. This study describes an important tool for quantifying egg deposition in a nondestructive manner, which is important in studying the colonization behavior and life history of insects of ecological and forensic importance.