This Partnerships for Innovation project from the University of Wisconsin-Milwaukee (UWM) focuses on innovation in the area of fast-acquisition-speed spectrally resolved two-photon microscopy (SR-TPM) and identification of biomarkers and labeled antibodies for use in research, diagnostics, and drug screening applications. As part of the PFI project, an Open Forum for Innovation (OFI) will be established to develop and guide proof-of-concept experiments in several areas related to SR-TPM technology development and applications: (1) high-spectral resolution and high-acquisition speed imaging scanners, (2) fiber laser alternatives to current solid-state lasers, and (3) novel biomarkers and labeled antibodies for detecting transient interactions between proteins in vivo, identifying rare cell types (such as circulating cancer cells) in body fluids, and developing living cell assays for cell signaling pathways to be used for drug screening.
The broader impacts of this research are (1) expansion of the SR-TPM customer base, currently represented by cellular biologists, to include both biochemists and drug researchers; (2) introduction of fiber laser products more broadly to the microscopy market; and (3) expansion of applications of fast-acquisition speed spectrally resolved fluorescence microscopy to include development of targeted diagnostics and therapeutics. This team's ability to address such needs would significantly expand the U.S. presence in the scanning fluorescent microscope market, by providing product capabilities that exceed those of existing products. Proof-of-concept experiments to be conducted will directly involve graduate, undergraduate students, and postdoctoral researchers who will work alongside consultants and lab scientists from the knowledge-enhancement partner companies. To add to the university's innovation capacity and create a cadre of UWM-grown scientist-entrepreneurs during the performance period of this project, OFI will evolve into a UWM University-Small Businesses Collaboratory (UWM-USBC), which will offer resources and networking opportunities to its member investigators, establish a gateway for businesses to UWM-developed technology, and promote the PFI project leaders as role models for knowledge transfer within UWM, in the Southeast Wisconsin area, and in the U.S.
Partners at the inception of the project include the Knowledge Enhancement Partnership (KEP) unit, consisting of UWM (College of Letters and Science (through its Department of Physics) and the Graduate School at the University of Wisconsin-Milwaukee), and three small businesses: Aurora Spectral Technologies, LLC (Milwaukee, WI), NeoClone Biotechnology International, LLC (Madison, WI), and PolarOnyx, Inc. (San Jose, CA). In addition, there are other core partners. These include private sector organizations: Brandt Innovative Technologies, Inc. (Pewaukee, WI) and the UWM Research Foundation, Inc. (UWMRF, Milwaukee, WI). In addition, two industrial collaborators and potential end users: MetriTrack, LLC (Hillside, IL) and AndroBioSys, Inc. (Buffalo, NY)], as well as academic collaborators and potential end users from the University of Toronto and University of Wisconsin-Madison, have already expressed strong interest in participating in the OFI.
Intellectual merit A partnership between researchers and product developers from several industrial and academic institutions has been assembled to tackle a long-standing problem of detecting, in living cells, the structure of molecular associations of membrane proteins. Capitalizing on microscopy technology for multi-color analysis of microscopic images of cells, originally dxeveloped at UW-Milwaukee, an Open Forum for Innovation (OFI) was established to develop and guide proof-of-concept experiments in several areas related to SR-TPM technology development and applications: (a) high-spectral resolution and high-acquisition speed imaging scanners, (b) fiber laser alternatives to current solid-state lasers used for two-photon microscopy, and (c) novel biomarkers and labeled antibodies for detecting transient interactions between proteins in vivo, and developing living cell assays for cell signaling pathways to be used for drug screening. To detect interactions between receptor proteins or between them and their ligands, a special exchange of energy is used—Förster resonance energy transfer (FRET)—that occurs when two fluorescent molecules, each attached to a protein of interest, come within a nanometer of each other. As a result of FRET, a laser-excited molecule, called a donor, transfers its energy to a nearby molecule, called the acceptor, which causes the acceptor to emit light. If the donor, which normally fluoresces green, is excited by laser light, but yellow is emitted by the acceptor, one knows that FRET occurred, and that the two proteins have interacted. The necessary color information is detected with exquisitely high-resolution with a laser-scanning microscope in which the spectrum of colors originating from a cellular sample containing the proteins of interest is projected by diffractive optics onto a fast and extremely sensitive camera. The partnership was formalized through establishment of a UWM-Small Businesses Collaboratory (http://www4.uwm.edu/collaboratory/index.cfm), of which both industrial and academic partners are active members. The whole body met twice for the duration of the grant in two-day symposia (http://www4.uwm.edu/letsci/conferences/microspectroscopy2013/) to review the state of the art of the technology and its applications in research, identify new directions of research and product-development opportunities, and establish smaller teams of investigators to pursue specific research projects. The research teams met and communicated as necessary for the duration of the entire projects. Broader impacts The new technology allows biomedical researchers to investigate cellular signaling under normal and pathological conditions, which holds the key to development and application of novel diagnostic assays and drugs to target specific diseases. In addition, the partnership between private industry and university researchers stimulates development of new intellectual property that can be quickly brought to market. This expands U.S. presence in in an area of technology development currently dominated by international companies (Zeiss, Nikon, Leica and Olympus). No other known technology has yet succeeded in detecting from measurements in living cells the spatial arrangement of proteins within a macromolecular complex (i.e., the quaternary structure of proteins). With further developments of this technology, it should also become possible to visualize in real time association and dissociation of proteins at their cellular location. Whenever a new technology allows one to visualize a biological process for the first time, it often leads to interesting and apriori unanticipated insights.
