Sequential color display for highly multiplexed in situ single-molecular detection
Abstract/Contents
- Abstract
- While much progress in high-throughput measurements of DNA, RNA and proteins has been made in vitro, our understanding of their dynamics within cells and tissues remains limited. By labeling each type of biomolecule with a unique color, in situ single-molecule visualization has allowed researchers to observe spatial context and copy numbers of those molecules within cells. However, utilizing one color per molecule is not enough for achieving a true multiplex analysis, as the number of colors is finite. To overcome this limitation, we developed a low-cost, highly accurate and rapid technology where each target DNA displays a specified color sequence via sequential hybridization of fluorescent DNA probes at several time points. Our sequential-color-display technology can be easily adapted to other DNA-based fluorescent detection methods to achieve highly multiplexed in situ detection of single DNA, RNA, and/or protein molecules in single cells. We were also able to combine our technology with a padlock-probe-based RNA detection method to detect multiple RNA species in situ with a 0.32% misidentification rate. The ability to measure genomic, transcriptional and proteomic variations in this manner may open an avenue to study the medical relevance of intercellular heterogeneity and intracellular molecular interactions. For example, this technique can be adapted to measure progression of cancers where malignant cells interact with nearby non-malignant cells, or diseases where infected or pathogenic cells interact with nearby healthy cells.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2017 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Peng, Yun-Chieh |
|---|---|
| Associated with | Stanford University, Department of Applied Physics. |
| Primary advisor | Byer, R. L. (Robert L.), 1942- |
| Primary advisor | Davis, Ronald W |
| Thesis advisor | Byer, R. L. (Robert L.), 1942- |
| Thesis advisor | Davis, Ronald W |
| Thesis advisor | Steinmetz, Lars |
| Advisor | Steinmetz, Lars |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Yun-Chieh Peng. |
|---|---|
| Note | Submitted to the Department of Applied Physics. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2017. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2017 by Yun-Chieh Peng
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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