Imaging alveolar membranes mimicking healthy and unfit lungs: the acquisition of a fluorescence microscope
Abstract
PUBLICLY RELEASABLE ABSTRACT. In this DURIP proposal, we describe critical research and educational aspects benefiting from the ins"tallation of the first Fluorescence Microscopy (FM) Imaging unit of the biomaterials facilities laboratory of the Materials Science and Engineering Building (MSEB) at the University of Illinois at Urbana-Champaign (UIUC). Numerous fluorescence imaging facilities are available at the UIUC campus within MSEB proximity including those at the Institute of Genome Biology (IGB) and the Beckman Ins"titute (BI). However, a vast number (ca 45) of Materials Science and Engineering (MatSE) students and postdocs, including those of t""he PI~s (Cecilia Leal) laboratory, require the daily use of FM techniques to investigate samples that should not be subjected to tra""nsporting between buildings. The acquisition of a FM unit will greatly benefit the research, training, and educational activities of"" about 5-10 MatSE faculty members conducting research in: 1) soft-materials self-assembly; 2) cellular delivery and nanomedicine, 3)"" stem-cell engineering, and 4) self-healing materials. All these topics are important research thrusts aligning with a number of dir"ectorates of the Department of Defense (DOD). This instrument will not only be beneficial for training and education of future scientists and engineers as it will foster research collaborations between MatSE faculty as well as other departments in close proximity to MSEB (e.g. Mechanical Engineering and Science). In this proposal the PI describes the particular need of FM techniques to be installed at MSEB to overcome a central hurdle in the research advance of a project sponsored by the Office of Naval Research (ONR) intended to underpin critical phase behavior of lung-mimetic membranes in mammals (marine and terrestrial) in unhealthy and diseased st"ates. This effort serves the ONR mission to ensure the health, fitness, and lifetime of marine mammals for duty. As opposed to terre""strial, diving mammals have lungs that comply with sudden lung collapse during diving. The mechanisms of lung compression/decompress"ion are strongly coupled with the properties of lipid membranes that stabilize a gas interface in alveoli. Our recent research efforts in this domain have led to the discovery of critical phase behavior of lungmimetic membranes under conditions of over-expression of certain lipids (cardiolipin) which happens to mammalian lungs afflicted with pneumonia. We elucidated that lung membranes displa"y a peculiar structural and dynamical behavior upon cardiolipin overexpression. Specifically, we have X-ray diffraction (SAXS) and s"olid-state Nuclear Magnetic Resonance (ssNMR) evidence that membranes become laterally heterogeneous and that those domains stack in three dimensions (3D) generating a 3D microphase phase separated multilamellar body. While SAXS and ssNMR convey important informat"ion at the nano-and-molecular scale, quantitative spatial resolution is still lacking. We propose the acquisition of a low maintenan""ce, long lasting, and highly modular multiphoton excitation fluorescence microscopy confocal set-up that will be coupled with an ele"ctroformation unit to produce Giant Unilamellar Vesicles (GUVs). The combination of the proposed FM unit with GUVs will enable 3D im"aging of membrane domains at the microscale as well as dynamics information of membrane heterogeneity formation, coarsening, and dis"solution.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 23, 2018
- Source ID
- N000141812087
Entities
People
- Cecilia Leal
Organizations
- Office of Naval Research
- United States Navy
- University of Illinois Urbana–Champaign