Multidimensional single-molecule imaging in live cells using total-internal-reflection fluorescence microscopy

We have developed a wide-field total-internal-reflection fluorescence microscope capable of imaging single molecules in live cells, resolved in both wavelength and polarization. We show fluorescence resonance energy transfer between single pairs of fluorescent molecules bound to signaling receptors...

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Bibliographic Details
Published inOptics letters Vol. 31; no. 14; p. 2157
Main Authors Webb, S E D, Needham, S R, Roberts, S K, Martin-Fernandez, M L
Format Journal Article
LanguageEnglish
Published United States 15.07.2006
Subjects
Animals
Cells, Cultured
Equipment Design
Equipment Failure Analysis
Fibroblasts - cytology
Fibroblasts - metabolism
Fluorescence Resonance Energy Transfer - instrumentation
Fluorescence Resonance Energy Transfer - methods
Fluorescent Dyes
Membrane Proteins - analysis
Mice
Microscopy, Fluorescence, Multiphoton - instrumentation
Microscopy, Fluorescence, Multiphoton - methods
Reproducibility of Results
Sensitivity and Specificity
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Summary:We have developed a wide-field total-internal-reflection fluorescence microscope capable of imaging single molecules in live cells, resolved in both wavelength and polarization. We show fluorescence resonance energy transfer between single pairs of fluorescent molecules bound to signaling receptors in the plasma membrane of live cells and demonstrate the importance of polarization discrimination in addition to wavelength separation.
ISSN:0146-9592
DOI:10.1364/OL.31.002157