Use of Internal Reflectance Infrared Spectroscopy for the in Situ Study of Supported Lipid Monolayers

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An instrument has been developed to acquire polarized attenuated total internal reflection (ATIR) mid-IR spectra of supported monomolecular films in situ at the lipid/water (L/W) interface. The monolayer film is applied to the treated surface of a trapezoidal germanium crystal as it is immersed into the subphase of a specially designed Langmuir-Blodgett trough. This approach offers several advantages over the study of monolayer films at the air/water (A/W) interface by external reflection techniques, including (1) an improved signal throughput and multiple reflections, both contributing to an improved signal-to-noise (S/N) ratio; (2) versatility with respect to the chemical nature of the surface; and (3) the ability to use standard spectral subtraction methods to compensate for absorption of the aqueous subphase. The enhanced sensitivity of this approach facilitates the study of in situ interactions of peptides with fully hydrated lipid membrane surfaces.

Keywords: FT-IR spectroscopy; Internal reflection spectroscopy; Langmuir-Blodgett technique; Lipid monolayers; Melittin

Document Type: Research Article


Affiliations: 1: Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6084 2: Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905 3: Hormel Institute, University of Minnesota, Austin, Minnesota 55912 4: Department of Chemistry, University of Georgia, Athens, Georgia 30602-2556

Publication date: April 1, 1995

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