AVS 52nd International Symposium
    Biomaterial Interfaces Wednesday Sessions
       Session BI2-WeM

Paper BI2-WeM11
Microspectroscopic Probing of Intracellular Structures by Observation of Infrared Linear Dichroism in Single Cells in a Micro-Fluidic Cuvette

Wednesday, November 2, 2005, 11:40 am, Room 312

Session: Biomembranes and Spectroscopy
Presenter: M. Schmidt, University of Maine
Authors: M. Schmidt, University of Maine
M. Rumpler, Max-Planck-Institut für Kolloid- und Grenzflächenforschung, Germany
N. Gierlinger, Max-Planck-Institut für Kolloid- und Grenzflächenforschung, Germany
U. Schade, BESSY GmbH, Germany
T. Rogge, Forschungszentrum Karlsruhe GmbH, Germany
P. Fratzl, Max-Planck-Institut für Kolloid- und Grenzflächenforschung, Germany
M. Grunze, University of Maine and Universität Heidelberg, Germany
Correspondent: Click to Email
Cellular properties and functions are closely related to cell structure. Probing intracellular structures and their dynamic nature is essential for the understanding of the functional characteristics of cells. Infrared (IR) microspectroscopy is an attractive tool for the investigation of biological materials and systems.@footnote 1@ Combining@footnote 2@ this technique with polarization modulation@footnote 3,4@ (PM) and employing synchrotron IR radiation allows us to perform polarization-dependent measurements with high spatial and temporal resolution. Thus, we are able to measure IR linear dichroism (LD) and hence determine preferred molecular orientation of distinct biochemical species in individual cells. Ultimately, observing single living cells in their native environment seems desirable when studying cell structure and function. Therefore, we developed an IR cuvette which facilitates the investigation of individual cells in aqueous solution. This custom-built, demountable and temperature-controllable micro-fluidic cuvette was microfabricated in order to meet the requirements of low pathlength (8 µm) and low volume (1 µL). Our goal is to gain insights into the formation and organization of the cytoskeleton in the context of cell adhesion. Using substrates with well defined surface properties and geometries we seek to control and model cell adhesion. Importantly, IR LD serves as an intrinsic marker for the preferred molecular orientation of the fibrous cytoskeletal proteins. Introduction of external stimuli such as chemicals, mechanical stress and substrate surface variation can be used to study the dynamic response and structural changes inside the cells. @FootnoteText@ @footnote 1@H.Y.N. Holman et al., J. Biomed. Opt. 7, 417 (2002).@footnote 2@Y. Shigematsu et al., Rev. Sci. Instrum. 72, 3927 (2001).@footnote 3@L.A. Nafie and M. Diem, Appl. Spectrosc. 33, 130 (1979).@footnote 4@T. Buffeteau et al., J. Chim. Phys. 90, 1467 (1993).