• cells cultures
• microcontact printing
• storm microscopy
• tessellation

Microcontact printing (μCP) is a lithographic technique used to create molecular substrates for various biological applications, including biosensors and scaffolds for cell growth. It transfers patterns onto substrates by depositing monolayers of targeted substances, making precise control and characterization of these patterns essential. This study aims to develop self-assembled protein patterns and innovative imaging tools for their nanoscale characterization under physiological conditions using super-resolution fluorescence microscopy, specifically STORM (Stochastic Optical Reconstruction Microscopy). STORM, part of the single-molecule localization microscopy (SMLM) family, surpasses traditional microscopy's diffraction limit, achieving nanometer-scale resolution, ideal for characterizing protein structures. The μCP and immunostaining protocols were optimized, and various parameters affecting the final image were evaluated. An innovative quantitative analysis tool based on spatial point patterns (SPP) was developed, using techniques like Voronoi tessellation and Delaunay triangulation for large protein network analysis. The study also explores the potential of using laminin scaffolds obtained via μCP as templates for studying cell-substrate interactions, including the observation of Human Intestinal Smooth Muscle Cells (HISMC) on laminin scaffolds and the development of an immunostaining protocol to analyze DNA distribution within the nucleus using STORM microscopy.