PUBLICATION
Optical lock-in detection imaging microscopy for contrast-enhanced imaging in living cells
- Authors
- Marriott, G., Mao, S., Sakata, T., Ran, J., Jackson, D.K., Petchprayoon, C., Gomez, T.J., Warp, E., Tulyathan, O., Aaron, H.L., Isacoff, E.Y., and Yan, Y.
- ID
- ZDB-PUB-081114-20
- Date
- 2008
- Source
- Proceedings of the National Academy of Sciences of the United States of America 105(46): 17789-17794 (Journal)
- Registered Authors
- Warp, Erica
- Keywords
- high-contrast, optical switches, “ac”-imaging, fluorescence microscopy
- MeSH Terms
-
- Actins
- Animals
- Cell Survival
- Cells, Cultured
- Fluorescent Dyes/chemistry
- Imaging, Three-Dimensional/methods*
- Mice
- Microscopy, Fluorescence
- Microscopy, Phase-Contrast/methods*
- Muscles/cytology
- NIH 3T3 Cells
- Neurons/cytology
- Rats
- Xenopus
- Zebrafish
- PubMed
- 19004775 Full text @ Proc. Natl. Acad. Sci. USA
Citation
Marriott, G., Mao, S., Sakata, T., Ran, J., Jackson, D.K., Petchprayoon, C., Gomez, T.J., Warp, E., Tulyathan, O., Aaron, H.L., Isacoff, E.Y., and Yan, Y. (2008) Optical lock-in detection imaging microscopy for contrast-enhanced imaging in living cells. Proceedings of the National Academy of Sciences of the United States of America. 105(46):17789-17794.
Abstract
One of the limitations on imaging fluorescent proteins within living cells is that they are usually present in small numbers and need to be detected over a large background. We have developed the means to isolate specific fluorescence signals from background by using lock-in detection of the modulated fluorescence of a class of optical probe termed "optical switches." This optical lock-in detection (OLID) approach involves modulating the fluorescence emission of the probe through deterministic, optical control of its fluorescent and nonfluorescent states, and subsequently applying a lock-in detection method to isolate the modulated signal of interest from nonmodulated background signals. Cross-correlation analysis provides a measure of correlation between the total fluorescence emission within single pixels of an image detected over several cycles of optical switching and a reference waveform detected within the same image over the same switching cycles. This approach to imaging provides a means to selectively detect the emission from optical switch probes among a larger population of conventional fluorescent probes and is compatible with conventional microscopes. OLID using nitrospirobenzopyran-based probes and the genetically encoded Dronpa fluorescent protein are shown to generate high-contrast images of specific structures and proteins in labeled cells in cultured and explanted neurons and in live Xenopus embryos and zebrafish larvae.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping