Bruker Atomic Force Microscope NanoWizard® 5

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Abstract In this Shared Instrumentation Grant proposal, we request funds to substantially enrich and expand capabilities of atomic force microscopy (AFM) based analysis of the single molecules, cells, and biomaterials at the University of Texas Health Science Center at San Antonio (UT Health SA) in the Bioanalytics and Single-Cell Core (BASiC). BASiC and its AFM laboratory is located in the Department of Molecular Medicine and it is a part of the Institutional Research Core under the Office of the Vice President for Research. The proposed advancement includes a purchase of the AFM system Nanowizard 5 from Bruker. The system will consist of a JPK NanoWizard® V AFM head, JPK Vortis 2.1 SPM Controller with its advanced version option, and a JPK HybridStage (200x200x200 µm³) mounted on a Nikon Eclipse Ti2-U inverted fluorescent microscope placed in an acoustic enclosure on a benchtop active vibration isolation system. Further enhancements include a microfluid system V2 FluidFM®, JPK Cell Mechanics Package and JPK SideView cantilever holder for indispensable for single cell manipulations. The system will be supported by a number of essential AFM hardware, accessories, tools, and specialized software for instruments control, data capture and analysis. The BASiC AFM facility has been successfully serving the University community and beyond for ten years delivering essential structural and mechanical data on cancer cells including patient isolated cells, organelles, macromolecules and biomaterials supporting numerous publications, grant applications and other associated activities. A broad array of basic, translational, and clinical studies at the UT Health SA depends on accessible, affordable, and customizable AFM-based molecular and cellular imaging technologies including mechanical phenotyping, chemical imaging and force spectrometry. The existing ten years old Nanoscope Catalyst is inefficient with the current overload of patient, animal model, and biomaterial samples leaving no free time for students training or critical methods development. Moreover, it cannot map medically crucial tissue, organoid and spacious samples under the physiologically relevant conditions. It is also severely restricted in studies of cell to cell interactions that are crucial for basic and medical studies. The current setup does not provide capabilities for single cell liquid injection, sampling, and manipulation vital for all sequencing and hyphenated applications interfaced with mechanical and fluorescent imaging. The Catalyst also shows a compromised resolution of molecular objects comparing to the proposed Nanowizard 5. The Catalyst cannot be upgraded and availability of its spare parts is limited. Therefore, the new proposed setup is indispensable not only to accommodate the quantity of current research and preserve its quality, but also to enormously expand its diversity and enhance its value that will benefit a broad spectrum of the biomedical users associated with the greater San Antonio scientific community.