Development and application of next generation lanthanide-based time-resolved luminescence microscopy
Abstract/Contents
- Abstract
- One of the fundamental experimental strategies of modern biology is "perturb and observe". Investigators perturb a complex biological system and, by observing the consequences of their experimental change, infer the function of the perturbed component(s). In order for researchers to be confident in their conclusions, perturbation and observation should have sufficient levels of specificity and sensitivity. Here, I describe two chemical technologies -- one observational and the other perturbational -- that enable better studies of biomolecules in whole-body organisms by making possible the selective visualization of lanthanide luminescence and the pharmacological control of protein stability. In chapter 1 and 2, I discuss a next-generation system for time-resolved lanthanide imaging. As luminescent lanthanide complexes have emission lifetimes that are > 100,000-fold longer than those of organic fluorophores, they can be selectively detected over fluorescent background by time-gating, enabling autofluorescence-free micrographs. However, the optical microscopy of lanthanide probes has not yet achieved detection limits and acquisition speed suitable for most biological applications. I describe three factors that have been impeding the efficient visualization of lanthanide luminescence, and present simple but powerful solutions to the respective problems. I also show novel imaging applications of lanthanide complexes that exploit unique chemical and photophysical properties of these organometallic probes. In chapter 3, I discuss the development of destabilizing domains (DDs) that allow rapid, reversible, and tunable control of protein levels in model organisms that thrive best at 20-25 °C. By genetically appending our DD to a protein of interest, the expression level of resulting fusion protein was successfully modulated by a small molecule ligand in C. elegans.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2016 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Cho, Ukrae | |
|---|---|---|
| Associated with | Stanford University, Department of Chemical and Systems Biology. | |
| Primary advisor | Chen, James Kenneth | |
| Thesis advisor | Chen, James Kenneth | |
| Thesis advisor | Lin, Michael Z | |
| Thesis advisor | Smolke, Christina D | |
| Thesis advisor | Wandless, Thomas | |
| Advisor | Lin, Michael Z | |
| Advisor | Smolke, Christina D | |
| Advisor | Wandless, Thomas |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Ukrae Cho. |
|---|---|
| Note | Submitted to the Department of Chemical and Systems Biology. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2016. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2016 by Ukrae Cho
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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