The development of super-resolution microscopy made it possible to surpass the diffraction limit of optical microscopy, enabling researchers to gain a nanometer scale understanding of cellular structures. While many applications have benefited from standard super-resolution microscopy, gaps remained making high-speed dynamic imaging in live cells impossible. To address this problem, single-point edge-excitation sub-diffraction (SPEED) microscopy was developed. This methodology enables the nanometer imaging of dynamic cell processes within live cells, the evaluation of subcellular structural information, the capacity to derive three-dimensional information from two-dimensional images within rotationally symmetric structures, and the interrogation of novel questions regarding the transport dynamics of macromolecules in a variety of cellular structures. Here, I have described the theory and method behind the current iteration of SPEED microscopy that we have developed and validated via Monte Carlo simulation. Further, a detailed description of how we have further developed SPEED microscopy to derive structural information within the nuclear pore complex as well as how SPEED has been applied to evaluate the export kinetics of mRNA. / Biology
| Identifer | oai:union.ndltd.org:TEMPLE/oai:scholarshare.temple.edu:20.500.12613/8497 |
| Date | January 2023 |
| Creators | Tingey, Mark, 0000-0002-0365-5585 |
| Contributors | Yang, Weidong, Dr., Liberles, David A., Habas, Raymond, Tuzel, Erkan |
| Publisher | Temple University. Libraries |
| Source Sets | Temple University |
| Language | English |
| Detected Language | English |
| Type | Thesis/Dissertation, Text |
| Rights | IN COPYRIGHT- This Rights Statement can be used for an Item that is in copyright. Using this statement implies that the organization making this Item available has determined that the Item is in copyright and either is the rights-holder, has obtained permission from the rights-holder(s) to make their Work(s) available, or makes the Item available under an exception or limitation to copyright (including Fair Use) that entitles it to make the Item available., http://rightsstatements.org/vocab/InC/1.0/ |
| Relation | http://dx.doi.org/10.34944/dspace/8461, Theses and Dissertations |
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