The development of new biophysical techniques is of paramount importance to the advancement of science. This proposal aims to advance existing spectroscopic imaging techniques to elucidate the understanding of cellular membranes and to further this understanding by developing both new model systems and techniques with unprecedented chemical and spatial resolution for application to biological systems. A Raman near fleld scanning optical microscope (NSOM) was developed during the K99 phase that will enable vibrational spectroscopic imaging with nanoscale spatial resolution. Vibrational spectroscopic imaging offers the opportunity to spatially resolve biological systems solely on the basis ofthe molecules present. This enables the study of the dynamical and organizational characteristics of cells and tissue without the need for external labels, which can disrupt the behavior of the system. This proposal aims to further develop and apply these new techniques to elucidate further the characteristics of biomolecules located within cellular membranes. Specifically, model systems will be utilized to explore the biophysical insights gained from this new methodology. Planar supported bilayers will be used as a model system for domain formation motifs, particularly the chemical composition and interactions that are found in lipid domains. l /Iore sophisticated model systems, such as confined vesicles, will be developed that will facilitate the ultimate goal of observing and understanding membrane heterogeneity and its role in cellular membrane processes such as signaling, and transport across the cell membrane. Ultimately, this proposal will utilize the established instrumentation and advance the methodology to gain insight into intact cell membranes. While domains are commonly ambiguously implicated in cell membrane processes related to pathogen infection, transport, and protein receptor activity, this research will demonstrate and clarify these molecular interactions at levels previously unattainable.

Public Health Relevance

An understanding ofthe behavior of cellular membranes is an integral component to Integrating the complex interactions that regulate cell behavior. This proposal seeks to develop new spectroscopic and biophysical methods, which will enable the direct investigation of cellular membranes at levels that are currently not feasible.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Research Transition Award (R00)
Project #
5R00RR024367-03
Application #
7892504
Study Section
Special Emphasis Panel (NSS)
Program Officer
Friedman, Fred K
Project Start
2009-07-15
Project End
2012-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
3
Fiscal Year
2010
Total Cost
$244,838
Indirect Cost
Name
University of Notre Dame
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Wang, Hao; Schultz, Zachary D (2013) The chemical origin of enhanced signals from tip-enhanced Raman detection of functionalized nanoparticles. Analyst 138:3150-7
Asiala, Steven M; Schultz, Zachary D (2013) Label-free in situ detection of individual macromolecular assemblies by surface enhanced Raman scattering. Chem Commun (Camb) 49:4340-2
Marr, James M; Schultz, Zachary D (2013) Imaging Electric Fields in SERS and TERS Using the Vibrational Stark Effect. J Phys Chem Lett 4:
Alexander, Kristen D; Schultz, Zachary D (2012) Tip-enhanced Raman detection of antibody conjugated nanoparticles on cellular membranes. Anal Chem 84:7408-14
Marr, James M; Li, Frank; Petlick, Alexandra R et al. (2012) The role of lateral tension in calcium-induced DPPS vesicle rupture. Langmuir 28:11874-80
Schultz, Zachary D (2011) Raman Spectroscopic Imaging of Cholesterol and Docosahexaenoic Acid Distribution in the Retinal Rod Outer Segment. Aust J Chem 64:611-616
Carrier, Stacey L; Kownacki, Corey M; Schultz, Zachary D (2011) Protein-ligand binding investigated by a single nanoparticle TERS approach. Chem Commun (Camb) 47:2065-7
Asiala, Steven M; Schultz, Zachary D (2011) Characterization of hotspots in a highly enhancing SERS substrate. Analyst 136:4472-9