Expansion microscopy and other tools for investigating the cell-material interface

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Surface topography on the scale of tens of nanometers to several micrometers substantially affects cell adhesion, migration, and differentiation. Recent studies using electron microscopy and super-resolution microscopy provide insight into how cells interact with surface nanotopography; however, the complex sample preparation and expensive imaging equipment required for these methods makes them not easily accessible. Expansion microscopy (ExM) is an affordable approach to image beyond the diffraction limit, but ExM cannot be readily applied to image the cell-material interface as most materials do not expand. Here, we develop a protocol that allows the use of ExM to resolve the cell-material interface with high resolution. We apply the technique to image the interface between U2OS cells and nanostructured substrates as well as the interface between primary osteoblasts with titanium dental implants. The high spatial resolution enabled by ExM reveals that although two proteins, such as AP2 and F-actin, both accumulate at curved membranes induced by vertical nanostructures, they are spatially segregated. We also employ upconversion nanoparticles as a better fluorophore to visualize proteins of interest in ExM. We believe that our protocol will enable the use of ExM as a powerful tool for cell-material interface studies. Additionally, we explore extracellular matrix fiber synthesis and proteomics as further tools to investigate the cell-material interface.


Type of resource text
Form electronic resource; remote; computer; online resource
Extent 1 online resource.
Place California
Place [Stanford, California]
Publisher [Stanford University]
Copyright date 2023; ©2023
Publication date 2023; 2023
Issuance monographic
Language English


Author Nakamoto, Melissa Lynn
Degree supervisor Cui, Bianxiao
Thesis advisor Cui, Bianxiao
Thesis advisor Bertozzi, Carolyn R, 1966-
Thesis advisor Solomon, Edward I
Degree committee member Bertozzi, Carolyn R, 1966-
Degree committee member Solomon, Edward I
Associated with Stanford University, School of Humanities and Sciences
Associated with Stanford University, Department of Chemistry


Genre Theses
Genre Text

Bibliographic information

Statement of responsibility Melissa Lynn Nakamoto.
Note Submitted to the Department of Chemistry.
Thesis Thesis Ph.D. Stanford University 2023.
Location https://purl.stanford.edu/dd914vs5034

Access conditions

© 2023 by Melissa Lynn Nakamoto

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