The accumulation of bacteria in surface attached biofilms, or biofouling, can be detrimental to human health, dental hygiene, and many industrial processes. A critical need in identifying and preventing the deleterious effects of biofilms is the ability to observe and quantify their development. Analytical methods capable of assessing early stage fouling are cumbersome or lab-confined, subjective, and qualitative. The software described in this invention disclosure applies a novel PNNL-developed image analysis algorithm to digital photos of fouled surfaces and provides the user with an objectively quantified measurement of fouling. The software and the accompanying sample preparation method will be described in a forthcoming peer-reviewed publication. Our aim in this invention disclosure is to make the source code available to researchers wishing to apply this method in their own research. Releasing the software is a critical step towards establishing Laboratory leadership in the biofouling research community by establishing new quantitative measurement standards.