 We have developed an efficient workflow for analyzing focal adhesions, FACE, in MDA-MB231 breast cancer cells. This workflow combines several ImageJ 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 Atko Horzum, Baron Osdall, and Devereen Pezzanotfer.