Summary (Core 1) High-throughput screening (HTS) has become the cornerstone of modern drug discovery both in Industry and Academia, allowing for the implementation of large-scale screening campaigns where thousands of compounds can be tested in search for novel drugs against a given disease. However, the overwhelming majority of existing HTS facilities and specialized equipment are set up in laboratories under low biosafety level (BSL) conditions. This makes it impossible to implement this approach for highly pathogenic microorganisms, such as Coccidioides spp., since work with these pathogens needs to be conducted inside high level biocontainment facilities. Thus, the overarching goal of this Drug Screening Core is to establish unique capabilities within BSL-3 conditions, which will allow for the implementation of fast, efficient and economical HTS techniques for large-scale screening of chemical libraries for the identification of novel anti-coccidioidal agents. More specifically, we propose to: i) develop low- and high-density microtiter plate-based models for Coccidioides spp. susceptibility testing amenable to HTS inside our BSL-3 facility, and ii) to fully develop a novel fungal cytological profiling (FCP) system based on imaging flow cytometry (IFC), and adapt it for drug screening and to determine the mode of action of novel drugs. Within the CCRC, this Core will support the screening and fungal cytological profiling activities in Project 1, provide services for down-selection of top candidates for in vivo evaluation in Project 2, and support Project 3 activities examining immunotherapeutic efficacy of antifungal drugs and vaccine candidates. Furthermore, once fully established, the technical capabilities of this Core will be provided to the Coccidioides scientific community at large, as well as to others in the field of Medical Mycology, and potentially other investigators working with BSL-3 organisms.