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Chin. Opt. Lett.
 Home  List of Issues    Issue 10 , Vol. 08 , 2010    10.3788/COL20100810.0947


Improving FRET efficiency measurement in confocal microscopy imaging
Huaina Yu1, Tongsheng Chen1, Junle Qu2
1 MOE Key Laboratory of Laser Life Science &
Institute of Laser Life Science, [South China Normal University], Guangzhou 510631, China
2 Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education &
Guangdong Province, Institute of Optoelectronics, [Shenzhen University], Shenzhen 518060, China

Chin. Opt. Lett., 2010, 08(10): pp.947-949-3

DOI:10.3788/COL20100810.0947
Topic:Medical optics and biotechnology
Keywords(OCIS Code): 170.1790  100.3008  110.2960  

Abstract
Spectral bleedthrough (SBT) ratio is dependent on the level of fluorescence intensity in confocal imaging. Precision Forster resonance energy transfer (FRET) algorithm corrects SBT ratio according to fluorescence intensity and avoids over- or under-estimation of SBT ratio. In this letter, we propose a new method to accurately measure the FRET efficiency of FRET plasmid in single living cells by combining the calculation of SBT in precision FRET algorithm with E-FRET formulae. We also use this method to measure the FRET efficiency of FRET-Bid, and find that in healthy A549 cells it is about 15%, which is verified by FRET acceptor photobleaching method.

Copyright: © 2003-2012 . This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Received:2010/6/22
Accepted:
Posted online:

Get Citation: Huaina Yu, Tongsheng Chen, Junle Qu, "Improving FRET efficiency measurement in confocal microscopy imaging," Chin. Opt. Lett. 08(10), 947-949-3(2010)

Note: This work was supported by the National Natural Science Foundation of China (Nos. 31071218 and 60627003) and the Natural Science Foundation of Guangdong Province (Nos. 8151063101000031 and 9251063101000009).



References

1. Y. Zhang, D. Xing, and L. Liu, Mol. Biol. Cell 20, 3007(2009).

2. G. Shan and W. Huang, Acta Opt. Sin. (in Chinese) 29, 1049 (2009).

3. X. Gao, D. Xing, and X. Gao, J. Cell Physiol. 219, 535(2009).

4. G. W. Gordon, G. Berry, X. H. Liang, B. Levine, and B. Herman, Biophys. J. 74, 2702 (1998).

5. L. Tron, J. Szollosi, S. Damjanovich, S. H. Helliwell, D. J. Arndt-Jovin, and T. M. Jovin, Biophys. J. 45, 939(1984).

6. M. G. Erickson, B. A. Alseikhan, B. Z. Peterson, and D. T. Yue, Neuron. 31, 973 (2001).

7. T. Zal and N. R. Gascoigne, Biophys. J. 86, 3923 (2004).

8. Y. Chen and A. Periasamy, J. Fluoresc. 16, 95 (2006).

9. M. Elangovan, H. Wallrabe, Y. Chen, R. N. Day, M. Barroso, and A. Periasamy, Methods 29, 58 (2003).

10. Y. Chen, M. Elangovan, and A. Periasamy, Molecular Imaging: FRET Microscopy and Spectroscopy (Oxford University Press, New York, 2005) p.126.

11. L. Wang, T. Chen, J. Qu, and X. Wei, J. Fluoresc. 20, 27 (2010).

12. Y. Zhang, D. Xing, and L. Liu, Chinese J. Lasers (in Chinese) 36, 2609 (2009).

13. J. Zhang, D. Xing, and X. Gao, J. Cell Physiol. 217, 518(2008).

14. F. Zhou, D. Xing, and W. R. Chen, Int. J. Cancer 125, 1380 (2009).

15. D. Sun, Q. Wu, C. Liu, H. Zhang, and L. Yao, Chinese J. Lasers (in Chinese) 35, 1409 (2008).


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