| AVS 58th Annual International Symposium and Exhibition | |
| Biomaterial Interfaces Division | Monday Sessions |
| Session BI-MoM |
| Session: | Biomolecules at Interfaces |
| Presenter: | Patrick Koelsch, Karlsruhe Institute of Technology (KIT), University of Heidelberg, Germany |
| Authors: | M.-O. Diesner, Karlsruhe Institute of Technology (KIT), University of Heidelberg, Germany P. Koelsch, Karlsruhe Institute of Technology (KIT), University of Heidelberg, Germany |
| Correspondent: | Click to Email |
The in situ monitoring of the interphase between a substrate and a cellular layer is of great interest as it allows determination of changes in surface properties and extracellular matrix (ECM) organization. The latter is an early indicator of major cellular processes like migration, adhesion, proliferation, metastasis, tissue formation, and gain or loss of differentiation occurring. Typically, ECM studies of adherent cells involve labeling and fixing cell samples, which may result in their disruption and in the loss of valuable information. In addition, the weak signal-to-noise ratio of fluorescent probes limits the probing capabilities at early stages of cell adhesion.
Recent work from our group has shown that sum-frequency-generation (SFG) spectroscopy can be used to interrogate the ordering of the ECM beneath adherent cells on an artificial substrate during these early stages.1,2 It turns out that SFG spectroscopy is suitable to probe the layer in between a solid substrate and living cells and that the information which can be obtained on such systems are twofold: first, changes of the surface coating can be investigated in real-time and in vitro on a molecular scale while cells adhere to it. Secondly, SFG spectroscopy is suitable for the determination of ordering parameters within the ECM without the need for labeling and processing.
In this contribution we will report on ordering phenomena occurring at early stages of rat and mouse embryonic fibroblasts adhesion on Au-coated Si wafers. Several phases observed during the adhesion process will be discussed and the results obtained by nonlinear optical spectroscopy will be correlated to classical tools including Western blot analysis of ECM constituents, fluorescent probes, and genetic screens blocking the formation of fibrils.
References
[1] M.-O. Diesner, C. Howell, V. Kurz, D. Verreault, and P. Koelsch. In vitro characterization of surface properties through living cells. J Phys Chem Lett, 1:2339– 2342, 2010.
[2] C. Howell, M.-O. Diesner, M. Grunze, and P. Koelsch. Probing the extracellular matrix with sum-frequency-generation spectroscopy. Langmuir, 24:13819–13821, 2008.