Oufti is a stand-alone open source software package designed to perform semi-automated sub-pixel precision detection and quantification of microbial cells from phase-contrast microscopy images. It can handle confluent cell samples and is scalable for high-throughput analysis of large datasets. The software also provides tools for quantitative analysis of both diffraction- and non-diffraction-limited fluorescent signals. Its GUI is interactive and includes post-processing functions for primary data analysis. Download Oufti.