Miniature and mass-producible fluorescence microscopes for biomedical imaging
Abstract/Contents
- Abstract
- Fluorescence microscopy has emerged as a workhorse of modern biology and medicine. Fluorescence microscopes are indispensable as tools for basic research in the biological sciences, enable platforms for drug discovery in the biotechnology and pharmaceutical industries, and are increasingly aiding in several clinical diagnostic applications. However, state-of-the-art fluorescence microscopes remain bulky and expensive benchtop instruments with an architecture that impedes usage in certain applications and a cost that precludes adoption in large numbers. For example existing benchtop fluorescence microscopes are not amenable for in vivo imaging in animals, with the mouse being a common animal subject, especially during awake, active behavior. For the field of neuroscience in particular, such an experimental capability permits correlating causal cellular processes with animal behavior -- a longstanding goal. Inspired by this need, we have designed a miniature fluorescence microscope that can be borne by a mouse during active behavior. We have fabricated several such microscopes, each less than 2 g in mass, perhaps heralding a transformation in fluorescence microscopy from today's bulky and expensive benchtop paradigm towards miniature and mass-producible devices. Our fabricated microscopes achieve 2.5 [mu]m spatial resolution imaging fields-of-view up to 800 [mu]m x 600 [mu]m at 36 Hz, suitable for cellular-level imaging at high temporal resolution. To facilitate design of our microscopes, we have adopted a modeling-based microscope design methodology, akin to that used in the design of integrated circuits, and have developed a set of tools to model the microscope as an integrated device from the specimen to final digital image. We have experimentally validated fabricated microscopes for in vivo brain imaging in freely behaving mice, specifically using them for: 1) Imaging cerebellar vasculature and hemodynamics during activity; and 2) Imaging cerebellar Purkinje cell Calcium dynamics, analyzing spiking activity of populations of neurons with single neuron specificity during different motor behaviors. We also show how several of these microscopes imaging in parallel can enable new high-throughput imaging solutions with the potential to achieve broad impact, specifically demonstrating high-throughput image-based assays for: 1) Mutant phenotype screening in genetic model species such as the zebrafish; and 2) Multiple well plate cell analyses. Finally, we use a Bayesian iterative image restoration algorithm to enhance acquired microscope images, and show how spatial resolution can be further scaled into the sub-micron regime by leveraging advances in digital image sensing technology in conjunction with our post-acquisition image restoration algorithm.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2010 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Ghosh, Kunal Kumar |
|---|---|
| Associated with | Stanford University, Department of Electrical Engineering |
| Primary advisor | El Gamal, Abbas A |
| Thesis advisor | El Gamal, Abbas A |
| Thesis advisor | Schnitzer, Mark Jacob, 1970- |
| Thesis advisor | Gambhir, Sanjiv Sam |
| Advisor | Schnitzer, Mark Jacob, 1970- |
| Advisor | Gambhir, Sanjiv Sam |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Kunal Kumar Ghosh. |
|---|---|
| Note | Submitted to the Department of Electrical Engineering. |
| Thesis | Thesis (Ph. D.)--Stanford University, 2010. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2010 by Kunal Kumar Ghosh
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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