PROJECT SUMMARY We are requesting a Leica STELLARIS 8 FALCON laser scanning confocal microscope for the microscopy facility in the Department of Molecular Microbiology and Immunology at the Johns Hopkins Bloomberg School of Public Health, which is part of the Johns Hopkins Medical Campus. A unique feature of our facility is the ability to perform live cell imaging experiments with biological agents that require Biosafety level 2 (BSL-2) containment, including human pathogens, such as Influenza A virus, Cryptococcus neoformans, Borrelia burgdorferi, Listeria monocytogenes, Plasmodium falciparum and Toxoplasma gondii. The proposed instrument will represent the first confocal microscope for this facility and within the current space restrictions will be singularly capable of supporting the various advanced confocal microscopy needs of our users. Notable features that convinced us during testing are its Wight Light Laser, translating into 350 laser lines, its imaging speed and the capacity to detect up to five different fluorophores simultaneously, while offering the gentle imaging conditions needed for live microscopy at confocal resolution. TauSense and FALCON enable the harnessing of fluorescence lifetime- based information and with this provide additional image quality, the ability to separate overlapping fluorophores and state-of-the-art measurements of protein interactions via FLIM-FRET. Altogether, the addition of this instrument to our facility will significantly enhance NIH-funded projects of our users and greatly contribute to the infectious disease focus and other key research areas of our institution.