Expansion Microscopy of Cells in Suspension
Expansion microscopy is a laboratory technique that enables nanoscale imaging of biological samples with conventional light microscopes. While expansion microscopy has traditionally been applied to specimens consisting of tissue and adherent cell culture, it has not been optimized for specimens cons...
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| Format: | Thesis |
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Massachusetts Institute of Technology
2023
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| Online Access: | https://hdl.handle.net/1721.1/151421 |
| _version_ | 1826217022983241728 |
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| author | Han, Nathan |
| author2 | Boyden, Edward Stuart |
| author_facet | Boyden, Edward Stuart Han, Nathan |
| author_sort | Han, Nathan |
| collection | MIT |
| description | Expansion microscopy is a laboratory technique that enables nanoscale imaging of biological samples with conventional light microscopes. While expansion microscopy has traditionally been applied to specimens consisting of tissue and adherent cell culture, it has not been optimized for specimens consisting of cells in suspension. In this work, a straightforward expansion microscopy protocol was developed for suspension cells. This protocol was validated across multiple cell types including in vitro and in vivo disease models, and multiple expansion microscopy versions encompassing different methods of sample fixation, anchoring, and gelation. Suspension cells imaged after conducting the protocol exhibited increased resolution compared to images of the initial raw sample, as well as a high rate of sample retention at a variety of initial concentrations. These findings suggest the potential for the wide use of expansion microscopy to study suspension cells, which provide a versatile and scalable system for investigating cellular processes and developing therapeutic treatments. The protocol created in this work can be directly used in the future to interrogate suspension cells at nanoscale resolution to identify underlying molecular and morphological mechanisms. |
| first_indexed | 2024-09-23T16:56:50Z |
| format | Thesis |
| id | mit-1721.1/151421 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T16:56:50Z |
| publishDate | 2023 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/1514212023-08-01T03:02:01Z Expansion Microscopy of Cells in Suspension Han, Nathan Boyden, Edward Stuart Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Expansion microscopy is a laboratory technique that enables nanoscale imaging of biological samples with conventional light microscopes. While expansion microscopy has traditionally been applied to specimens consisting of tissue and adherent cell culture, it has not been optimized for specimens consisting of cells in suspension. In this work, a straightforward expansion microscopy protocol was developed for suspension cells. This protocol was validated across multiple cell types including in vitro and in vivo disease models, and multiple expansion microscopy versions encompassing different methods of sample fixation, anchoring, and gelation. Suspension cells imaged after conducting the protocol exhibited increased resolution compared to images of the initial raw sample, as well as a high rate of sample retention at a variety of initial concentrations. These findings suggest the potential for the wide use of expansion microscopy to study suspension cells, which provide a versatile and scalable system for investigating cellular processes and developing therapeutic treatments. The protocol created in this work can be directly used in the future to interrogate suspension cells at nanoscale resolution to identify underlying molecular and morphological mechanisms. M.Eng. 2023-07-31T19:38:30Z 2023-07-31T19:38:30Z 2023-06 2023-06-06T16:35:15.449Z Thesis https://hdl.handle.net/1721.1/151421 In Copyright - Educational Use Permitted Copyright retained by author(s) https://rightsstatements.org/page/InC-EDU/1.0/ application/pdf Massachusetts Institute of Technology |
| spellingShingle | Han, Nathan Expansion Microscopy of Cells in Suspension |
| title | Expansion Microscopy of Cells in Suspension |
| title_full | Expansion Microscopy of Cells in Suspension |
| title_fullStr | Expansion Microscopy of Cells in Suspension |
| title_full_unstemmed | Expansion Microscopy of Cells in Suspension |
| title_short | Expansion Microscopy of Cells in Suspension |
| title_sort | expansion microscopy of cells in suspension |
| url | https://hdl.handle.net/1721.1/151421 |
| work_keys_str_mv | AT hannathan expansionmicroscopyofcellsinsuspension |