Simultaneous widefield single molecule orientation and FRET microscopy in cells
Optics Express, Vol. 16, Issue 25, pp. 20258-20265 (2008)
http://dx.doi.org/10.1364/OE.16.020258
Enhanced HTML
Acrobat PDF (267 KB)
Abstract
We combine single molecule fluorescence orientation imaging with single-pair fluorescence resonance energy transfer microscopy, using a total internal reflection microscope. We show how angles and FRET efficiencies can be determined for membrane proteins at the single molecule level and provide data from the epidermal growth factor receptor system in cells.
© 2008 Optical Society of America
OCIS Codes
(000.1430) General : Biology and medicine
(180.2520) Microscopy : Fluorescence microscopy
ToC Category:
Medical Optics and Biotechnology
History
Original Manuscript: September 10, 2008
Revised Manuscript: November 12, 2008
Manuscript Accepted: November 12, 2008
Published: November 24, 2008
Virtual Issues
Vol. 4, Iss. 2 Virtual Journal for Biomedical Optics
Citation
S. E. D. Webb, D. J. Rolfe, S. R. Needham, S. K. Roberts, D. T. Clarke, C. I. McLachlan, M. P. Hobson, and M. L. Martin-Fernandez, "Simultaneous widefield single molecule orientation and FRET microscopy in cells," Opt. Express 16, 20258-20265 (2008)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-16-25-20258
Sort: Year | Journal | Reset
References
- J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, "Protein structural dynamics by single-molecule fluorescence polarization," Prog. Biophys. Mol. Biol. 74, 1-35 (2000). [CrossRef] [PubMed]
- J. N. Forkey, M. E. Quinlan, M. A. Shaw, J. E. T. Corrie, and Y. E. Goldman, "Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization," Nature 422, 399-404 (2003). [CrossRef] [PubMed]
- E. A. Jares-Erijman and T. M. Jovin, "FRET imaging," Nat. Biotechnol. 21, 1387-1395 (2003). [CrossRef] [PubMed]
- T. Ha, "Single-molecule fluorescence resonance energy transfer," Methods 25, 78-86 (2001). [CrossRef] [PubMed]
- M. Tokunaga, K. Kitamura, K. Saito, A. H. Iwane, and T. Yanagida, "Single molecule imaging of fluorophores and enzymatic reactions achieved by objective-type total internal reflection fluorescence microscopy," Biochem. Biophys. Res. Comm. 235, 47-53 (1997). [CrossRef] [PubMed]
- S. E. D. Webb, S. R. Needham, S. K. Roberts, and M. L. Martin-Fernandez, "Multidimensional single-molecule imaging in live cells using total-internal-reflection fluorescence microscopy," Opt. Lett. 31, 2157-2159 (2006). [CrossRef] [PubMed]
- S. Hohng, C. Joo, and T. Ha, "Single-molecule three-color FRET," Biophys. J. 87, 1328-1337 (2004). [CrossRef] [PubMed]
- J. Schlessinger, "Cell signaling by receptor tyrosine kinases," Cell 103, 211-225 (2000). [CrossRef] [PubMed]
- S. E. D. Webb, S. K. Roberts, S. R. Needham, C. J. Tynan, D. J. Rolfe, M. D. Winn, D. T. Clarke, R. Barraclough, and M. L. Martin-Fernandez, "Single-molecule imaging and fluorescence lifetime imaging microscopy show different structures for high- and low-affinity epidermal growth factor receptors in A431 cells," Biophys. J. 94, 808-819 (2008). [CrossRef]
- R. S. Chandran, S. E. John, and M. Amit, "Using fluorescence resonance energy transfer to measure distances along individual DNA molecules: Corrections due to nonideal transfer," J. Chem. Phys. 122, 061103 (2005). [CrossRef]
Cited By |
OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.
Multimedia
| Multimedia Files | Recommended Software |
| » Media 1: MPG (4834 KB) | QuickTime |
| » Media 2: MPG (3354 KB) | QuickTime |





OSA is a member of 