This proposal describes the Live Cell Microscopy Core for the Midwest Regional Center of Excellence forBiodefense and Emerging Infectious Disease Research (MRCE). This Core is to provide MRCE, otherregional centers and greater research community the ability to perform a variety of advanced fluorescencemicroscopy techniques with live BSL2 and BSL3 organisms. Before the facility was available, there was noregional option for performing advanced microscopy techniques on BSL3 organisms. The technicalinstrumentation and scientific expertise are also readily available to the public health community in the eventof a bioterror, outbreak or pandemic event. Two fluorescence microscopes and a Laser-CaptureMicrodissection (LCM) microscope are available in the facility. The Nikon LiveScan Swept-Field Confocalmicroscope supports multi-dimensional data acquisition (x, y, z, time, wavelengths and multi-point), and theTILL Wide-Field microscope allows studies of ratio imaging (ions and pH) and FRET (FluorescenceResonance Energy Transfer). Their super light sensitivity and video speed minimize photobleaching andphototoxicity, which is crucial for long-term imaging of fluorescent live samples. Cells are kept viable in amicroscope cage-incubator that enables control of temperature, humidity and gas concentrations. Such livemotionimaging can be applied to explore dynamic localization, colocalization, trafficking, interaction andturnover between pathogen and cells, cellular organelles, proteins or ions. The LCM microscope enablesdetailed examination of extracted DMA, RNA and protein from targeted portions of histological sections.Perspective users will apply to the Core, and specific experimental protocols will be designed by the user inconjunction with the core PI. Biohazard safety issues will be resolved in conjunction with Dr. Susan Cook ofEnvironmental Safety at Washington University, and appropriate clearances will be obtained in advance.Continued funding of this Core is required to increase the availability of cutting-edge microscopy forinvestigators in research community and in the event of an emergency to greatly advance our understandingbiological mechanisms of pathogens.
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