3D expansion microscopy on nanostructure arrays to visualize the nano-bio Interfaces in super resolution

• Main Author: Huang, Lizhen
• Other Authors: Zhao Wenting
• Format: Thesis-Master by Research
• Language: English
• Published: Nanyang Technological University 2024
• Subjects: Engineering; Super-resolution; 3D calibration; Nano structure; Expansion microscopy; Nano-bio interface; Membrane curvature
• Online Access: https://hdl.handle.net/10356/174088
• Institution: Nanyang Technological University

The concept of bio interface encompasses the intricate interaction between biological entities and surfaces, where diverse surfaces are intentionally engineered to modulate cellular responses and scrutinize associated protein activities. Many of these crucial cellular reactions and protein responses unfold at the nanometer scale, presenting a challenge in visualization due to the diffraction limit. This project introduces an innovative approach by leveraging high-resolution Expansion Microscopy (ExM) coupled with 3D calibration to delve into the intricate nano-bio interface. The main hypothesis of my thesis is that nanostructure arrays characterized by controllable geometry and patterns can serve as reliable references in three dimensions (3D) for the calibration of fluorescence signals at the nano-bio interface following hydrogel expansion in Expansion Microscopy (ExM). High-resolution and high-accuracy imaging is essential for comprehending any responses or changes. Using ExM combined with calibration, the study successfully maps the nanoscale distribution of key podosome proteins around arrays of nanostructures, characterizing their colocalization not only in 2D but also in 3D within the z plane. This innovative calibration in the ExM approach emerges as a potent tool to enhance the accuracy of super-resolution images, thereby facilitating a more profound understanding of nanoscale cellular structures.