Abstract
Physical interfaces mediate interactions between multiple types of cells. Despite the importance of such interfaces to the cells’ function, their high-resolution optical imaging has been typically limited due to poor alignment of the interfaces relative to the optical plane of imaging. Here, we present a simple and robust method to align cell-cell interfaces in parallel to the coverslip by adhering the interacting cells to two opposing coverslips and bringing them into contact in a controlled and stable fashion. We demonstrate aberration-free high-resolution imaging of interfaces between live T cells and antigen-presenting cells, known as immune synapses, as an outstanding example. Imaging methods may include multiple diffraction-limited and super-resolution microscopy techniques (e.g., bright-field, confocal, STED, and dSTORM). Thus, our simple and widely compatible approach allows imaging with high- and super-resolution the intricate structure and molecular organization within a variety of cell-cell interfaces.
Original language | American English |
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Title of host publication | The Immune Synapse |
Editors | Cosima T. Baldari, Michael L. Dustin |
Publisher | Humana Press Inc. |
Pages | 149-158 |
Number of pages | 10 |
ISBN (Electronic) | 978-1-0716-3135-5 |
ISBN (Print) | 978-1-0716-3134-8, 978-1-0716-3137-9 |
DOIs | |
State | Published - 2023 |
Publication series
Name | Methods in Molecular Biology |
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Volume | 2654 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Bibliographical note
Publisher Copyright:© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Keywords
- Cell-to-cell interaction
- Diffraction limit
- Imaging
- Immune synapse
- Microcopy
- Super-resolution
- T cells