Calabuig-Barroso, Alejandro (2016) DEVELOPMENT OF ADVANCED INTERFEROMETRIC TECHNICHES FOR THE STUDY OF CELL-MATERIAL INTERACTION. [Tesi di dottorato]

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Abstract

Recently, the development of innovative materials able to control and guide the cellular responses for specific applications has been produced a great interest. The study of cell-material interaction is important because it provides a better understanding of how they affect the physical and chemical properties of materials on the cellular dynamics. In fact, it is known that some types of signals regulate important processes such as adhesion, differentiation, and other cellular events. Currently, it has extended the use of highly advanced analytical devices for the study of these phenomena. For morphological analysis using the fluorescence microscope that allows you to report different types of subcellular structures such as the actin cytoskeleton and the cell nucleus. In addition, the atomic force microscope, allows the analysis of the topography of biomaterials before being used as platforms for cell adhesion. The disadvantage in using these techniques is the one hand, that the fluorescent microscope uses of the dyes to analyze the morphology of the cell and, on the other hand, the atomic force microscope does not allow the real-time analysis of topographical formation of although the material has a high spatial resolution. The thesis is targeted to develop three types of instruments based optical interferometry, to provide new types of methods in cell-material interaction study, for both the manipulation and characterization of biomaterials, both for the quantitative study of cell behaviour under dynamic changes of the physical properties of adhesion substrates. The experimental apparatus for the characterization of the optical lithographic processes provides the capability of real-time monitor the formation in azopolimeri of so-called "Surface Relief Gratings" (SRG). In addition, via a special digital holographic microscope, the dynamics of fibroblasts has been studied on a petri glass platform. Experiments show that this device allow to open a new way of research on the quantitative analysis of cellular damage caused by the effect of phototoxicity induced by electromagnetic waves. Moreover, morphological processes of necrotic death can be monitored with quantitative-temporal phase analysis. Finally, a microscopic device based on the analysis surface in "Total Internal Reflection" has been improved through advanced interferometric techniques to obtain quantitative information about the cell adhesion processes. Validation and application of the technique have been demonstrated.

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