Abstract

Spatial and temporal information about intracellular objects and their dynamics within a living cell are essential for dynamic analysis of such objects in cell biology. A specific intracellular object can be discriminated by photoactivatable fluorescent proteins that exhibit pronounced light-induced spectral changes. Here, we report on selective labeling and tracking of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. We performed selective labeling of a single mitochondrion in a living tobacco BY-2 cell using two-photon photoconversion of Kaede. Using this technique, we demonstrated that, in plants, the directed movement of individual mitochondria along the cytoskeletons was mediated by actin filaments, whereas microtubules were not required for the movement of mitochondria. This single-organelle labeling technique enabled us to track the dynamics of a single organelle, revealing the mechanisms involved in organelle dynamics. The technique has potential application in direct tracking of selective cellular and intracellular structures.

© 2007 Optical Society of America

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  1. J. Lippincott-Schwartz and G. H. Patterson, "Development and use of fluorescent protein markers in living cells," Science 300, 87-91 (2003).
    [CrossRef] [PubMed]
  2. G. H. Patterson and J. Lippincott-Schwartz, "A photoactivatable GFP for selective photolabeling of proteins and cells," Science 297, 1873-1877 (2002).
    [CrossRef] [PubMed]
  3. J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
    [CrossRef] [PubMed]
  4. M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
    [CrossRef] [PubMed]
  5. M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
    [CrossRef] [PubMed]
  6. D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
    [CrossRef]
  7. D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
    [CrossRef]
  8. Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
    [CrossRef]
  9. J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
    [CrossRef] [PubMed]
  10. N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
    [CrossRef]
  11. R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
    [CrossRef] [PubMed]
  12. H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
    [CrossRef]
  13. H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
    [CrossRef] [PubMed]
  14. S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
    [CrossRef] [PubMed]
  15. S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
    [CrossRef]
  16. T. Nagata, Y. Nemoto, and S. Hasezawa, "Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants," Int. Rev. Cyt. 132, 1-30 (1992).
    [CrossRef]
  17. M. Wada and N. Suetsugu, "Plant organelle positioning," Curr. Opin. Plant Biol. 7, 626-631 (2004).
    [CrossRef] [PubMed]
  18. A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
    [CrossRef] [PubMed]
  19. A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
    [CrossRef] [PubMed]
  20. K. Van Gastel, R. H. Köhler, and J.-P. Vervelen, "Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules" J. Exp. Bot. 53, 659-667 (2002).
    [CrossRef]
  21. U. K Tirlapur and K. König, "Targeted transfection by femtosecond laser," Nature 418, 290-291 (2002).
    [CrossRef] [PubMed]
  22. W. Watanabe, N. Arakawa, S. Matsunaga, T. Higashi, K. Fukui, K. Isobe, and K. Itoh, "Femtosecond laser disruption of subcellular organelles in a living cell," Opt. Express 12, 4203-4213 (2004),
    [CrossRef] [PubMed]
  23. A. Heisterkamp, I. Z. Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar, and D. E. Ingber,   "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005)
    [CrossRef] [PubMed]
  24. M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
    [CrossRef] [PubMed]
  25. T. Shimada, W. Watanabe, S. Matsunaga, T. Higashi, H. Ishii, K. Fukui, K. Isobe, and K. Itoh, "Intracellular disruption of mitochondria in living HeLa cells with a 76-MHz femtosecond laser oscillator," Opt. Express 13, 9869-9880 (2005)
    [CrossRef] [PubMed]
  26. A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
    [CrossRef]
  27. K. König, "Laser tweezers and multiphoton microscopes in life sciences," Histochem. Cell Biol. 114, 79-92 (2000).
    [PubMed]

2006

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

2005

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
[CrossRef] [PubMed]

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

A. Heisterkamp, I. Z. Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar, and D. E. Ingber,   "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005)
[CrossRef] [PubMed]

T. Shimada, W. Watanabe, S. Matsunaga, T. Higashi, H. Ishii, K. Fukui, K. Isobe, and K. Itoh, "Intracellular disruption of mitochondria in living HeLa cells with a 76-MHz femtosecond laser oscillator," Opt. Express 13, 9869-9880 (2005)
[CrossRef] [PubMed]

2004

W. Watanabe, N. Arakawa, S. Matsunaga, T. Higashi, K. Fukui, K. Isobe, and K. Itoh, "Femtosecond laser disruption of subcellular organelles in a living cell," Opt. Express 12, 4203-4213 (2004),
[CrossRef] [PubMed]

M. Wada and N. Suetsugu, "Plant organelle positioning," Curr. Opin. Plant Biol. 7, 626-631 (2004).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

2003

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

J. Lippincott-Schwartz and G. H. Patterson, "Development and use of fluorescent protein markers in living cells," Science 300, 87-91 (2003).
[CrossRef] [PubMed]

2002

G. H. Patterson and J. Lippincott-Schwartz, "A photoactivatable GFP for selective photolabeling of proteins and cells," Science 297, 1873-1877 (2002).
[CrossRef] [PubMed]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

K. Van Gastel, R. H. Köhler, and J.-P. Vervelen, "Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules" J. Exp. Bot. 53, 659-667 (2002).
[CrossRef]

U. K Tirlapur and K. König, "Targeted transfection by femtosecond laser," Nature 418, 290-291 (2002).
[CrossRef] [PubMed]

2000

K. König, "Laser tweezers and multiphoton microscopes in life sciences," Histochem. Cell Biol. 114, 79-92 (2000).
[PubMed]

1999

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

1992

T. Nagata, Y. Nemoto, and S. Hasezawa, "Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants," Int. Rev. Cyt. 132, 1-30 (1992).
[CrossRef]

Aida, G. P.

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

Akatsuka, M.

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

Ando, R.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

Arakawa, N.

Arimura, S.

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

Arndt-Jovin, D. J.

J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
[CrossRef] [PubMed]

Arnoult, D.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Barozzi, S.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Belousov, V. V.

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

Ben-Yakar, A.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Chan, D. C.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Chen, H.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Chen, Y.

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

Chepurnykh, T. V.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

Chisholm, A. D.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Chudakov, D. M.

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

Cinar, H.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Cinar, H. N.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Diaspro, A.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Dunahay, T. G.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Faretta, M.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Fradkov, A. F.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

Frohlick, J. A.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Fukui, K.

Furuta, T.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

Gallagher, L. A.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Gurskaya, N. G.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

Hama, H.

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

Hasezawa, S.

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

T. Nagata, Y. Nemoto, and S. Hasezawa, "Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants," Int. Rev. Cyt. 132, 1-30 (1992).
[CrossRef]

Heisterkamp, A.

Higashi, T.

Hoshino, H.

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

Hüttman, G.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

Ikura, M.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

Ingber, D. E.

Ishii, H.

Isobe, K.

Itoh, K.

Ivanchenko, S.

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Jin, Y.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Karasawa, S.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

Karbowski, M.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Köhler, R. H.

K. Van Gastel, R. H. Köhler, and J.-P. Vervelen, "Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules" J. Exp. Bot. 53, 659-667 (2002).
[CrossRef]

König, K.

U. K Tirlapur and K. König, "Targeted transfection by femtosecond laser," Nature 418, 290-291 (2002).
[CrossRef] [PubMed]

K. König, "Laser tweezers and multiphoton microscopes in life sciences," Histochem. Cell Biol. 114, 79-92 (2000).
[PubMed]

Kumagai, F.

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

Kumar, S.

Kumar Mal, T.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

Lidke, K. A.

J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
[CrossRef] [PubMed]

Lippincott-Schwartz, J.

J. Lippincott-Schwartz and G. H. Patterson, "Development and use of fluorescent protein markers in living cells," Science 300, 87-91 (2003).
[CrossRef] [PubMed]

G. H. Patterson and J. Lippincott-Schwartz, "A photoactivatable GFP for selective photolabeling of proteins and cells," Science 297, 1873-1877 (2002).
[CrossRef] [PubMed]

Lukyanov, K. A

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

Lukyanov, K. A.

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

Lukyanov, S.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

Macdonald, P. J.

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

Matsunaga, S.

Maxwell, I. Z.

Mazur, E.

Mazurkiewicz, A. M.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Meehl, J. B.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Miyawaki, A.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

Mizuno, H.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

Müller, J. D.

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

Nagata, T.

T. Nagata, Y. Nemoto, and S. Hasezawa, "Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants," Int. Rev. Cyt. 132, 1-30 (1992).
[CrossRef]

Nakazono, M.

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

Nebenführ, A.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Nemoto, Y.

T. Nagata, Y. Nemoto, and S. Hasezawa, "Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants," Int. Rev. Cyt. 132, 1-30 (1992).
[CrossRef]

Nickerson, J. A.

Nienhaus, G. U.

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Nienhaus, U.

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

Noack, J.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

Novoselov, V. V.

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

Nukina, N.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

Oswald, F.

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Paltauf, G.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

Patterson, G. H.

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

J. Lippincott-Schwartz and G. H. Patterson, "Development and use of fluorescent protein markers in living cells," Science 300, 87-91 (2003).
[CrossRef] [PubMed]

G. H. Patterson and J. Lippincott-Schwartz, "A photoactivatable GFP for selective photolabeling of proteins and cells," Science 297, 1873-1877 (2002).
[CrossRef] [PubMed]

Post, J. N.

J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
[CrossRef] [PubMed]

Rieger, B.

J. N. Post, K. A. Lidke, B. Rieger, and D. J. Arndt-Jovin, "One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos," FEBS Lett. 579,325-330 (2005).
[CrossRef] [PubMed]

Rocker, C.

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Röcker, C.

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

Salih, A.

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Schmitt, F.

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Schneider, M.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Shcheglov, A. S.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

Shimada, T.

Shimizu, H.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

Skinner, J. P.

Y. Chen, P. J. Macdonald, J. P. Skinner, G. H. Patterson, and J. D. Müller, "Probing nucleocytoplasmic transport by two-photon activation of PA-GFP," Microscopic Res. Tech. 69, 220-226 (2006).
[CrossRef]

Smith, C. L.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Souslova, E. A.

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

Spindler, K. D.

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Staehelin, L. A.

A. Nebenführ, L. A. Gallagher, T. G. Dunahay, J. A. Frohlick, A. M. Mazurkiewicz, J. B. Meehl, and L. A. Staehelin, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system," Plant Physiol. 121, 1127-1141 (1999).
[CrossRef] [PubMed]

Staroverov, D. B.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

Suetsugu, N.

M. Wada and N. Suetsugu, "Plant organelle positioning," Curr. Opin. Plant Biol. 7, 626-631 (2004).
[CrossRef] [PubMed]

Testa, I.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Tirlapur, U. K

U. K Tirlapur and K. König, "Targeted transfection by femtosecond laser," Nature 418, 290-291 (2002).
[CrossRef] [PubMed]

Tong, K. I.

H. Mizuno, T. Kumar Mal, K. I. Tong, R. Ando, T. Furuta, M. Ikura, and A. Miyawaki, "Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein," Mol. Cell 12, 795-1058 (2003).
[CrossRef]

Tsutsui, H.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
[CrossRef] [PubMed]

Tsutsumi, N.

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

Underwood, J. M.

Van Gastel, K.

K. Van Gastel, R. H. Köhler, and J.-P. Vervelen, "Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules" J. Exp. Bot. 53, 659-667 (2002).
[CrossRef]

Verkhusha, V. V.

N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov,T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov, and K. A Lukyanov, "Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light," Nature Biotechnol. 24, 461-465 (2006).
[CrossRef]

D. M. Chudakov, V. V. Verkhusha, D. B. Staroverov, E. A. Souslova, S. Lukyanov, and K. A. Lukyanov, "Photoswitchable cyan fluorescent protein for protein tracking," Nature Biotechnol. 22, 1435-1439 (2004).
[CrossRef]

Vervelen, J.-P.

K. Van Gastel, R. H. Köhler, and J.-P. Vervelen, "Plant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules" J. Exp. Bot. 53, 659-667 (2002).
[CrossRef]

Vogel, A.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

Wada, M.

M. Wada and N. Suetsugu, "Plant organelle positioning," Curr. Opin. Plant Biol. 7, 626-631 (2004).
[CrossRef] [PubMed]

Watanabe, W.

Wiedenmann, J.

S. Ivanchenko, C. Röcker, F. Oswald, J. Wiedenmann, and U. Nienhaus, "Targeted green-red photoconversion of EosFP, a fluorescent marker protein," J. Biol. Phys. 31, 249-259 (2005).
[CrossRef]

J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Rocker, A. Salih, K. D. Spindler, G. U. Nienhaus, "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion," Proc Natl. Acad. Sci. USA 101, 15905-15910 (2004).
[CrossRef] [PubMed]

Yamamoto, J.

S. Arimura, J. Yamamoto, G. P. Aida, M. Nakazono, and N. Tsutsumi, "Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution," Proc. Natl. Acad. Sci. USA,  101, 7805-7808 (2004).
[CrossRef] [PubMed]

Yamamoto-Hino, M.

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein," Proc. Natl. Acad. Sci. USA,  99, 12651-12656 (2002).
[CrossRef] [PubMed]

Yanik, M. F.

M. F. Yanik, H. Cinar, H. N. Cinar, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 822-822 (2004).
[CrossRef] [PubMed]

Yoneda, A.

A. Yoneda, M. Akatsuka, H. Hoshino, F. Kumagai, and S. Hasezawa, "Decision of spindle poles and division plane by double preprophase bands in a BY-2 cell line expressing GFP-tubulin," Plant Cell Physiol. 46, 531-538 (2005).
[CrossRef] [PubMed]

Youle, R. J.

M. Karbowski, D. Arnoult, H. Chen, D. C. Chan, C. L. Smith, and R. J. Youle, "Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis," J. Cell Biol. 164, 493-499 (2004).
[CrossRef] [PubMed]

Zaraisky, A. G.

D. M. Chudakov, V. V. Belousov, A. G. Zaraisky, V. V. Novoselov, D. B. Staroverov, D. B., Zorov, S. Lukyanov, and K. A. Lukyanov, "Kindling fluorescent proteins for precise in vivo photolabeling," Nature Biotechnol. 21, 191-194 (2003).
[CrossRef]

Appl. Phys. B

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," Appl. Phys. B 81, 1015-1047 (2005).
[CrossRef]

Biophys J.

M. Schneider, S. Barozzi, I. Testa, M. Faretta, and A. Diaspro, "Two-photon activation and excitation properties of PA-GFP in the 720-920 nm region," Biophys J. 89, 1346-1352 (2005).
[CrossRef] [PubMed]

Curr. Opin. Plant Biol.

M. Wada and N. Suetsugu, "Plant organelle positioning," Curr. Opin. Plant Biol. 7, 626-631 (2004).
[CrossRef] [PubMed]

EMBO Rep.

H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina, and A. Miyawaki, "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter," EMBO Rep. 6, 233-238 (2005).
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Supplementary Material (1)

» Media 1: AVI (2142 KB)     

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Figures (6)

Fig. 1.
Fig. 1.

Schematic diagram of the experimental setup for single-organelle labeling and tracking. FI, faraday isolator; M, mirror; P, SF10 prism; L, lens; ND, neutral density filter; DM, dichroic mirror; GM, pair of galvanometer mirrors; OB, objective lens; BP, bandpass filter; PMT, photomultiplier tube.

Fig. 2.
Fig. 2.

Selective photoconversion of mitochondria with Kaede in a fixed BY-2 cell. Left column, stacked 3D confocal images of green fluorescence obtained through bandpass filter (BP; 510 to 540 nm); middle column, stacked 3D confocal images of red fluorescence obtained through bandpass filter (560 to 600 nm); right column, white light transmission images (A) before femtosecond laser pulse irradiation and (B) after femtosecond laser pulse irradiation. A single mitochondrion was photoconverted from green to red by 750-nm femtosecond laser pulses with an energy of 0.053 nJ/pulse (exposure time: 1 s). A target mitochondrion is indicated by the yellow arrow. Scale bar: 10 μm.

Fig. 3.
Fig. 3.

Tracking of a mitochondrion in a living BY-2 cell. Scale bar: 10 μm. Time-lapse stacked confocal images along z axis (0.5-μm steps, total 13 slices) after marking a mitochondrion by two-photon excitation. The movement of the mitochondrion labeled by two-photon conversion could be tracked for 5 hours. The trajectory of the labeled mitochondrion is shown by the yellow line. See supplementary movie (2.1 MB). [Media 1]

Fig. 4.
Fig. 4.

3D Mitochondrial tracking in a living BY-2 cell. (a) 3D mitochondrial tracking. (b) Projections of 3D tracks on XY, XZ, YZ planes are presented. 2D and 3D (grid squares are 5μm×5 μm) depiction of representative mitochondria track. The trajectory of the labeled mitochondrion is shown in successive image stacks (yellow line).

Fig. 5.
Fig. 5.

The movement and instantaneous velocity of mitochondria whose cells were treated with the actin-polymerization inhibitor bistheonellide A or the tubulin-polymerization inhibitor propyzamide. (a) The position of a mitochondrion. Projections of 3D tracks are presented (grids are 50 μm). (b) Variation of instantaneous velocity of individual mitochondria in control cells over 100 s. The plots were taken at intervals of 1.13 s.

Fig. 6.
Fig. 6.

Effect of actin and microtubule polymerization inhibitor on average mitochondrial velocity. Average velocity of a mitochondrion (*; vs actin polymerization inhibitor, p<0.001). Cells were treated with the actin-polymerization inhibitor bistheonellide A or the tubulin-polymerization inhibitor propyzamide. The average velocity without any treatment (control) is also indicated. Error bars indicate the standard error of the mean.

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