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.

© 2010 Chinese Optics Letters

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

2010

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

2009

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

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

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

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

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

2008

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

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

2006

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

2004

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

2003

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

2001

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

1998

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

1984

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

Alseikhan, B. A.

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

Arndt-Jovin, D. J.

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

Barroso, M.

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

Berry, G.

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

Chen, T.

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

Chen, W. R.

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

Chen, Y.

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

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

Damjanovich, S.

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

Day, R. N.

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

Elangovan, M.

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

Erickson, M. G.

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

Gao, X.

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

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

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

Gascoigne, N. R.

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

Gordon, G. W.

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

Helliwell, S. H.

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

Herman, B.

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

Huang, W.

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

Jovin, T. M.

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

Levine, B.

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

Liang, X. H.

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

Liu, C.

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

Liu, L.

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

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

Periasamy, A.

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

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

Peterson, B. Z.

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

Qu, J.

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

Shan, G.

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

Sun, D.

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

Szollosi, J.

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

Tron, L.

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

Wallrabe, H.

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

Wang, L.

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

Wei, X.

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

Wu, Q.

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

Xing, D.

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

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

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

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

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

Yao, L.

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

Yue, D. T.

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

Zal, T.

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

Zhang, H.

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

Zhang, J.

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

Zhang, Y.

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

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

Zhou, F.

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

Acta Opt. Sin. (in Chinese)

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

Biophys. J.

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

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

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

Chinese J. Lasers (in Chinese)

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

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

Int. J. Cancer

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

J. Cell Physiol.

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

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

J. Fluoresc.

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

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

Methods

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

Mol. Biol. Cell

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

Neuron.

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

Other

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

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