The need for new antibiotics that are active against multidrug resistant pathogens has created renewed interest in discovering antibiotics from sources of natural products. We have developed a new approach, bacterial cytological profiling (BCP), that uses quantitative fluorescence microscopy to measure the effects of antibiotic treatment on individual cells. Antibiotics that target different cellular pathways and different steps within a pathway generate unique cytological profiles, allowing identification of the likely MOA of new compounds within a few hours. BCP is especially useful for identification of natural products in crude natural product extracts because it rapidly discriminates between antibiotics with different mechanisms of action (MOA). The BCP screening platform can detect nuisance compounds and multiple activities in partially purified or crude natural product extracts and it can detect active molecules at sub-inhibitory concentrations. The goal of this project is to develop a comprehensive natural product identification platform that combines BCP with genome mining methods to rapidly identify novel molecules and their biosynthetic gene clusters. Linnaeus Bioscience Inc. is a start up company founded to commercialize this technology and make it accessible on a fee for service basis to the pharmaceutical industry and the scientific community. This proposals describes a set of specific aims that will allow us to develop a simple, robust platform for discovering novel natural products active against drug resistant bacteria.