Development of two-photon polymerization-based protocols for the investigation of neuronal mechanobiology

Abstract

Fundamental neuronal studies are concerned with the investigation and understanding of the behaviour of cells in response to certain chemical or physical cues. The field of study concerned with physical cues and their effect on neurons is referred to as neuronal mechanobiology or neuromechanobiology. These cues include properties such as the geometry (i.e. 2D vs. 3D substrates), topography (i.e. roughness of the substrate), and stiffness of the substrate, which can affect migration, phenotypic expression, and neurite outgrowth of neurons. In recent years, the study of mechanobiology has witnessed substanrial developments due to our increasing ability to fabricate (physiologically) relevant environments that simulate specific features of the native extracellular matrix (ECM). Specifically, within micro and nano-fabrication techniques, two-photon polymerization (2PP) gained a lot of traction and proved to be a versarile method for the fabrication of microstructures to be used in mechanobiological studies. The technology of 2PP allows indeed the fabrication of specifically designed complex 3D microstructures with a resolution that can reaches down to 50 nm in feature size in a reproducible manner with relative ease.
In this dissertation, I present multiple microfabrication and processing protocols developed specifically for mechanobiological studies of neurons and microglia (part of the glial cells category), two of the most abundant cell types in the brain.