Characterization of cytogels using acousto-microscopy-based oscillating rod rheometry

Juergen Bereiter-Hahn; Oliver Wagner

doi:10.1117/12.434178

24 July 2001 Characterization of cytogels using acousto-microscopy-based oscillating rod rheometry

Juergen Bereiter-Hahn, Oliver Wagner

Proceedings Volume 4335, Advanced Nondestructive Evaluation for Structural and Biological Health Monitoring; (2001) https://doi.org/10.1117/12.434178
Event: 6th Annual International Symposium on NDE for Health Monitoring and Diagnostics, 2001, Newport Beach, CA, United States

Abstract

The physical properties of cytoplasm are primarily determined by the state of cytoskeletal element, i.e. their polymerisation, crosslinking and supramolecular interactions with other molecules. These interactions are involved in signal transduction processes as well as in morphogenesis. Scanning acoustic microscopy proved to be a powerful tool to determine the mechanical properties of living cells. The interpretation of the sound propagation parameters, however, has to be based on investigation of in vitro models. Therefore polymerisation of actin and tubulin have been followed using a novel oscillating rod rheometer which allows for synchronous determination of sound velocity, sound attenuation and viscosity. Sound velocity measures the elastic propterties of cytogels, attenuation the supramolecular associations. All these parameters are evaluated with minimal strain, in the range of 1- 100 nm actin with glycolytic enzymes not only modulated polymerisation in a specific, and substrate dependent manner, but also the stiffness of the fibrils was altered, e.g. by hexokinase in the presence of high ATP, this enzyme exhibited actin severing properties and reduced stiffness. Differences in polymerisation kinetics were observed comparing pyrene-labeled actin fluorimetry and oscillating rod viscosimetry. This comparison led to the detection of pseudocrystalline structures produced by g-actin and aldolase (in the absence of fructose-bisphophate, the substrate of aldolase). Elastic stiffness of actin filaments can be modulated by ATP/ADP and by actin binding proteins (e.g. the glycolytic enzyme hexokinase) as well. The in vitro observations support the interpretation of SAM data calculated for living cells.

Citation Download Citation

Juergen Bereiter-Hahn and Oliver Wagner "Characterization of cytogels using acousto-microscopy-based oscillating rod rheometry", Proc. SPIE 4335, Advanced Nondestructive Evaluation for Structural and Biological Health Monitoring, (24 July 2001); https://doi.org/10.1117/12.434178

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