 We have developed an efficient workflow for analyzing focal adhesions, FACE, in NDAMB231 breast cancer cells. This workflow combines several image A plugins to simplify the process and reduce false negatives. It also allows for the detection of FACE with minimal background interference. This approach can be used to quantitatively analyze other fluorescent images containing localized or focal signals in a high background environment. This article was authored by Acahorsum, Baron Osdall, and Devrim Pezzanotfer.