(DURIP) Atomic Force Microscope for Single-Molecule Investigations of Biomolecular Actuation by Ion-Responsive Repeat Proteins
Abstract
Danielle J. Mai, an Assistant Professor of Chemical Engineering at Stanford University, is pursuing support for an atomic force microscope (AFM) that will be coupled to a fluorescence optical-microscope. The instrument will be a major upgrade to an aging BioAFM that is nearing the end of its service life; this instrument is currently housed at the Cell Sciences Imaging Facility at Stanford University. The proposed instrument is a Bruker-JPK Nano-Wizard V BioAFM, which will be integrated with an existing Zeiss Axio Observer Z1 inverted optical microscope to enable simultaneous fluorescence imaging during force measurements. The optical microscope will also be upgraded with new optical components suited for low-light and single-molecule fluorescence imaging, which will uniquely enable simultaneous multi-mode single-molecule imaging and force measurements. The Nano-Wizard V will accelerate active AFOSR-funded research of PI Mai that focuses on engineering ion-responsive repeat proteins for soft actuating materials. This research involves multi-scale, multi-mode characterization of protein-based materials including single protein molecules, hydrogels, and soft actuator devices. Features of the proposed Nano Wizard V include faster measurement speeds to increase the throughput of single-molecule force spectroscopy experiments, an expanded z range to establish force-extension relationships for high molecular-weight polypeptides, simultaneous fluorescence imaging to correlate molecular-scale forces with genetically encoded fluorescence probes, and an improved user interface to reduce experimental setup time and fast-track training for new users. In addition to advancing research in the Mai Lab, the Nano Wizard V will expand AFM imaging capabilities at Stanford University to critically enable research in functional materials, polymer science and engineering, and the biomedical sciences. This instrument s integrated fluorescence optical microscopy capabilities will be unique across campus; this feature will enable state-of-the art correlative optical imaging with maps of surface topography, nano-mechanics, and-or electrical properties. High-resolution imaging and high-precision force measurements enable research ranging from single-molecule biophysics to mechanobiology of single cells, tissues, and materials. No other shared user facility in Northern California offers BioAFM, so we anticipate users from local peer institutions across the San Francisco Bay Area and Northern California.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Feb 05, 2025
- Source ID
- FA95502410063
Entities
People
- Danielle J Mai
Organizations
- Air Force Office of Scientific Research
- Stanford University
- United States Air Force