Electro-optic techniques for nanosecond imaging and applications to fluorescence lifetime microscopy
Abstract/Contents
- Abstract
- Fluorescence microscopy allows targeted visualization of biological matter across spatial and temporal scales. Most methods rely on measurements of image intensity that are taken with a standard camera sensor at low frame rates. The nanosecond excited state lifetime of a fluorescent probe also carries valuable information about the local environment which can be used to improve optical measurements, but it cannot be captured with common cameras. Current lifetime detectors are either too slow or too noisy for many applications. This thesis presents the development of nanosecond imaging optics and the electro-optic fluorescence lifetime imaging microscopy (EO-FLIM) method. Fast optical gating of a wide-field image is achieved using electro-optic crystals, allowing lifetimes to be measured on scientific camera sensors with high sensitivity. EO-FLIM improves photon throughput by several orders of magnitude over standard time-resolved detectors. Lifetime is estimated from a ratio of optical intensities, which increases the information content of the captured image while also rejecting intensity noise and motion artifacts. Several optical systems and technological improvements are presented which have enabled wide-field lifetime imaging of single fluorescent molecules, combination of lifetime imaging with super-resolution localization microscopy, and lifetime recording of neuron action potentials and sub-threshold voltage activity in vivo at kilohertz frame rates. Applications to light-sheet microscopy and time-of-flight imaging are also shown.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2023; ©2023 |
| Publication date | 2023; 2023 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Bowman, Adam Joseph |
|---|---|
| Degree supervisor | Kasevich, Mark |
| Thesis advisor | Kasevich, Mark |
| Thesis advisor | Moerner, William |
| Thesis advisor | Schnitzer, Mark |
| Degree committee member | Moerner, William |
| Degree committee member | Schnitzer, Mark |
| Associated with | Stanford University, School of Humanities and Sciences |
| Associated with | Stanford University, Department of Applied Physics |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Adam J. Bowman. |
|---|---|
| Note | Submitted to the Department of Applied Physics. |
| Thesis | Thesis Ph.D. Stanford University 2023. |
| Location | https://purl.stanford.edu/vm973kq1178 |
Access conditions
- Copyright
- © 2023 by Adam Joseph Bowman
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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