Measuring conformational dynamics of biological molecules with single-molecule tracking and fluorescence correlation spectroscopy
Abstract/Contents
- Abstract
- The past two decades have seen great advances in single-molecule techniques capable of probing biomolecular systems with spatial resolution down to the nanometer level. However few techniques are specifically designed to measure larger scale organization between a few tens and a few hundreds of nanometers, especially when other figures of merit are considered, such as high time-resolution or the possibility to do measurements in bulk and without immobilization. Such features would be particularly useful in the study of molecular structures like chromatin or other nucleoprotein complexes, that exhibit organizational details and dynamics across a broad range of time and length scales. I present how the combination of fluorescence correlation spectroscopy (FCS) and single molecule tracking can be used to measure the conformational dynamics of molecules that diffuse freely in solution, at the scale of a few tens of nanometers. I discuss some aspects of the instrumentation as well as the parameters that define the spatial and temporal resolution. I describe other advantages of the technique such as the possibility to bypass a common difficulty associated with fluorescence correlation measurements and obtain a signal that is not convolved with the blinking dynamics of the fluorescent probe itself. Finally I present an experimental proof-of-principle where tracking-FCS was applied to measure the end-to-end Brownian dynamics of short DNA fragments in the semilflexible regime.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2014 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Limouse, Charles |
|---|---|
| Associated with | Stanford University, Department of Applied Physics. |
| Primary advisor | Mabuchi, Hideo |
| Thesis advisor | Mabuchi, Hideo |
| Thesis advisor | Spakowitz, Andrew James |
| Thesis advisor | Straight, Aaron, 1966- |
| Advisor | Spakowitz, Andrew James |
| Advisor | Straight, Aaron, 1966- |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Charles Limouse. |
|---|---|
| Note | Submitted to the Department of Applied Physics. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2014. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2014 by Charles Bruno Maurice Limouse
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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