Expansion microscopy and other tools for investigating the cell-material interface
Abstract/Contents
- Abstract
- 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.
Description
| 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 |
Creators/Contributors
| 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 |
Subjects
| 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
- Copyright
- © 2023 by Melissa Lynn Nakamoto
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