Project Summary: The current proposal outlines a strategic plan to implement novel technologies within containment that will advance the NIAID mission to support pandemic preparedness and biomedical research on risk group III (RGIII) pathogens. This proposal focuses on the implementation of cutting edge equipment to advance in vitro (Aim1), in vivo (Aim2), and molecular investigation (Aim3) of high containment pathogens and also seeks to build upon the diagnostic expertise of the Scientific Director to establish a high containment molecular infrastructure capable of rapidly responding to future emerging pandemic threats with rapid diagnostic test development and real-world implementation for patient care and public health intervention. Innovations include: 1) First proposal seeking to acquire cutting edge equipment to increase scientific productivity at University of Alabama at Birmingham (UAB)’s SEBLAB, 2) Novel use of the Zeiss lattice light-sheet microscope within containment, 3) Innovative implementation of ultra-high resolution computed tomography and PET/SPECT capabilities enabling use of novel radionuclides generated by the nation’s largest TR24 cyclotron (UAB) for non-invasive diagnostic imaging research on RGIII pathogens, 4) Create the ability to perform bioenergetic analyses and single cell sequencing at SEBLAB, and 5) Implement molecular infrastructure within containment enabling rapid diagnostic test development and validation with a clear path to implementation in CLIA/CAP-accredited clinical and public health laboratories. Given the current pandemic, risks for future outbreaks, and rapid advancement of research technologies, investment in the UAB SEBLAB as outlined in the current proposal will translate into real-world impacts on pandemic preparedness and accelerate hyper-competitive basic biomedical and translational research output that advances our basic understanding and biodefense capabilities against high containment pathogens.