Abstract

We present a combination of nonlinear microscopy and optical trapping applied to three-dimensional imaging and manipulation of intracellular structures in living cells. We use Titanium-sapphire laser pulses for nonlinear microscopy of the nuclear envelope and the microtubules marked with green fluorescent protein in fission yeast. The same laser source is also used to trap small lipid granules naturally present in the cell. The trapped granule is used as a handle to exert a pushing force on the cell nucleus. The granule is moved in a raster-scanning fashion to cover the area of the nucleus and hence displace the nucleus away from its normal position in the center of the cell. Such indirect manipulations of an organelle (e.g., nucleus) can be useful when direct trapping of the chosen organelle is disadvantageous or inefficient. We show that nonlinear microscopy and optical manipulation can be performed without substantial damage or heating of the cell. We present this method as an important tool in cell biology for manipulation of specific structures, as an alternative to genetic and biochemical methods. This technique can be applied to several fundamental problems in cell biology, including the mechanism of nuclear positioning and the spatial coordination of nuclear and cell division.

© 2005 Optical Society of America

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  1. W. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
    [CrossRef] [PubMed]
  2. W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Bio-technol. 21, 1369–1377 (2003).
    [CrossRef]
  3. W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).
  4. M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
    [CrossRef] [PubMed]
  5. S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
    [CrossRef] [PubMed]
  6. D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
    [CrossRef] [PubMed]
  7. A. Ashkin, J. M. Dziedzic, J. E. Bjorkholm, S. Chu, “Observation of a single-beam gradient force optical trap for dielectric particles,” Opt. Lett. 11, 288–290 (1986).
    [CrossRef] [PubMed]
  8. A. Ashkin, “History of optical trapping and manipulation of small-neutral particle, atoms, and molecules,” IEEE J. Sel. Top. Quantum Electron. 6, 841–856 (2000).
    [CrossRef]
  9. A. Ashkin, J. M. Dziedzic, T. Yamane, “Optical trapping and manipulation of single cells using infrared laser beams,” Nature 330, 769–771 (1987).
    [CrossRef] [PubMed]
  10. A. Ashkin, J. M. Dziedzic, “Optical trapping and manipulation of viruses and bacteria,” Science 235, 1517–1520 (1987).
    [CrossRef] [PubMed]
  11. M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
    [CrossRef] [PubMed]
  12. A. Ashkin, J. M. Dziedzic, “Internal cell manipulation using infrared laser traps,” Proc. Natl. Acad. Sci. U.S.A. 86, 7914–7918 (1989).
  13. M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).
  14. T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).
  15. M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).
  16. K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
    [CrossRef] [PubMed]
  17. D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
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  18. D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
    [PubMed]
  19. L. Sacconi, E. Froner, R. Antolini, M. R. Taghizadeh, A. Choudhury, F. S. Pavone, “Multiphoton multifocal microscopy exploiting a diffractive optical element,” Opt. Lett. 28, 1918–1920 (2003).
    [CrossRef] [PubMed]
  20. L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
    [CrossRef]
  21. K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).
  22. M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
    [CrossRef]
  23. S. P. Alexander, C. L. Rieder, “Chromosome motion during attachment to the vertebrate spindle: initial saltatory-like behavior of chromosomes and quantitative analysis of force production by nascent kinetochore fibers,” J. Cell Biol. 113, 805–815 (1991).
  24. S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
    [CrossRef] [PubMed]
  25. E. K. McCully, C. F. Robinow, “Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy,” J. Cell Sci. 9, 475–507 (1971).
    [PubMed]
  26. I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
    [CrossRef]
  27. E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).
  28. K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).
  29. D. R. Drummond, R. A. Cross, “Dynamics of interphase microtubules in Schizosaccharomyces pombe,” Curr. Biol. 10, 766–775 (2000).
    [CrossRef] [PubMed]
  30. F. Chang, P. Nurse, “How fission yeast fission in the middle,” Cell 84, 191–194 (1996).
    [CrossRef] [PubMed]
  31. Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
    [CrossRef] [PubMed]
  32. S. Reinsch, P. Gonczy, “Mechanisms of nuclear positioning,” J. Cell Sci. 111, 2283–2295 (1998).
    [PubMed]
  33. I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”
  34. A. Sievers, L. Heyder-Caspers, “The effect of centrifugal accelerations on the polarity of statocytes and on the graviperception of cress roots,” Planta 157, 64–70 (1983).
    [CrossRef] [PubMed]
  35. I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
    [CrossRef] [PubMed]

2005 (1)

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

2004 (3)

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

2003 (5)

D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
[CrossRef] [PubMed]

L. Sacconi, E. Froner, R. Antolini, M. R. Taghizadeh, A. Choudhury, F. S. Pavone, “Multiphoton multifocal microscopy exploiting a diffractive optical element,” Opt. Lett. 28, 1918–1920 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Bio-technol. 21, 1369–1377 (2003).
[CrossRef]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).

2002 (3)

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
[CrossRef] [PubMed]

2000 (4)

D. R. Drummond, R. A. Cross, “Dynamics of interphase microtubules in Schizosaccharomyces pombe,” Curr. Biol. 10, 766–775 (2000).
[CrossRef] [PubMed]

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

A. Ashkin, “History of optical trapping and manipulation of small-neutral particle, atoms, and molecules,” IEEE J. Sel. Top. Quantum Electron. 6, 841–856 (2000).
[CrossRef]

1999 (1)

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

1998 (3)

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

S. Reinsch, P. Gonczy, “Mechanisms of nuclear positioning,” J. Cell Sci. 111, 2283–2295 (1998).
[PubMed]

1997 (2)

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

1996 (1)

F. Chang, P. Nurse, “How fission yeast fission in the middle,” Cell 84, 191–194 (1996).
[CrossRef] [PubMed]

1993 (1)

K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
[CrossRef] [PubMed]

1992 (1)

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

1991 (1)

S. P. Alexander, C. L. Rieder, “Chromosome motion during attachment to the vertebrate spindle: initial saltatory-like behavior of chromosomes and quantitative analysis of force production by nascent kinetochore fibers,” J. Cell Biol. 113, 805–815 (1991).

1990 (1)

W. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

1989 (2)

A. Ashkin, J. M. Dziedzic, “Internal cell manipulation using infrared laser traps,” Proc. Natl. Acad. Sci. U.S.A. 86, 7914–7918 (1989).

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

1987 (2)

A. Ashkin, J. M. Dziedzic, T. Yamane, “Optical trapping and manipulation of single cells using infrared laser beams,” Nature 330, 769–771 (1987).
[CrossRef] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical trapping and manipulation of viruses and bacteria,” Science 235, 1517–1520 (1987).
[CrossRef] [PubMed]

1986 (1)

1983 (1)

A. Sievers, L. Heyder-Caspers, “The effect of centrifugal accelerations on the polarity of statocytes and on the graviperception of cress roots,” Planta 157, 64–70 (1983).
[CrossRef] [PubMed]

1971 (1)

E. K. McCully, C. F. Robinow, “Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy,” J. Cell Sci. 9, 475–507 (1971).
[PubMed]

Aist, J. R.

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

Alexander, S. P.

S. P. Alexander, C. L. Rieder, “Chromosome motion during attachment to the vertebrate spindle: initial saltatory-like behavior of chromosomes and quantitative analysis of force production by nascent kinetochore fibers,” J. Cell Biol. 113, 805–815 (1991).

Andrews, J. J.

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Antolini, R.

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

L. Sacconi, E. Froner, R. Antolini, M. R. Taghizadeh, A. Choudhury, F. S. Pavone, “Multiphoton multifocal microscopy exploiting a diffractive optical element,” Opt. Lett. 28, 1918–1920 (2003).
[CrossRef] [PubMed]

Ashkin, A.

A. Ashkin, “History of optical trapping and manipulation of small-neutral particle, atoms, and molecules,” IEEE J. Sel. Top. Quantum Electron. 6, 841–856 (2000).
[CrossRef]

A. Ashkin, J. M. Dziedzic, “Internal cell manipulation using infrared laser traps,” Proc. Natl. Acad. Sci. U.S.A. 86, 7914–7918 (1989).

A. Ashkin, J. M. Dziedzic, T. Yamane, “Optical trapping and manipulation of single cells using infrared laser beams,” Nature 330, 769–771 (1987).
[CrossRef] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical trapping and manipulation of viruses and bacteria,” Science 235, 1517–1520 (1987).
[CrossRef] [PubMed]

A. Ashkin, J. M. Dziedzic, J. E. Bjorkholm, S. Chu, “Observation of a single-beam gradient force optical trap for dielectric particles,” Opt. Lett. 11, 288–290 (1986).
[CrossRef] [PubMed]

Bacri, J. C.

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

Ballerini, R.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Bergman, K.

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

Berg-Sørensen, K.

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

Berns, M. W.

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Bjorkholm, J. E.

Block, S. M.

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
[CrossRef] [PubMed]

Cahalan, M. D.

M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
[CrossRef] [PubMed]

Capitanio, M.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Chadd, E. H.

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

Chang, F.

F. Chang, P. Nurse, “How fission yeast fission in the middle,” Cell 84, 191–194 (1996).
[CrossRef] [PubMed]

Chikashige, Y.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

Choudhury, A.

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

Chu, S.

Cohen, M.

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

Cross, R. A.

D. R. Drummond, R. A. Cross, “Dynamics of interphase microtubules in Schizosaccharomyces pombe,” Curr. Biol. 10, 766–775 (2000).
[CrossRef] [PubMed]

de Ruijter, N. C.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Denk, W.

W. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Ding, D. Q.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

Dogterom, M.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Drummond, D. R.

D. R. Drummond, R. A. Cross, “Dynamics of interphase microtubules in Schizosaccharomyces pombe,” Curr. Biol. 10, 766–775 (2000).
[CrossRef] [PubMed]

Dunlap, D.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Dziedzic, J. M.

A. Ashkin, J. M. Dziedzic, “Internal cell manipulation using infrared laser traps,” Proc. Natl. Acad. Sci. U.S.A. 86, 7914–7918 (1989).

A. Ashkin, J. M. Dziedzic, T. Yamane, “Optical trapping and manipulation of single cells using infrared laser beams,” Nature 330, 769–771 (1987).
[CrossRef] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical trapping and manipulation of viruses and bacteria,” Science 235, 1517–1520 (1987).
[CrossRef] [PubMed]

A. Ashkin, J. M. Dziedzic, J. E. Bjorkholm, S. Chu, “Observation of a single-beam gradient force optical trap for dielectric particles,” Opt. Lett. 11, 288–290 (1986).
[CrossRef] [PubMed]

Emons, A. M.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Esseling, J. J.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Faivre-Moskalenko, C.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Finzi, L.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Froner, E.

Gelles, J.

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

Gittes, F.

E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).

Giuntini, M.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Goldberg, M.

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

Gonczy, P.

S. Reinsch, P. Gonczy, “Mechanisms of nuclear positioning,” J. Cell Sci. 111, 2283–2295 (1998).
[PubMed]

Grier, D. G.

D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
[CrossRef] [PubMed]

Grierson, C. S.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Gruenbaum, Y.

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

Guillen, N.

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

Haraguchi, T.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

Harel, A.

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

Heyder-Caspers, L.

A. Sievers, L. Heyder-Caspers, “The effect of centrifugal accelerations on the polarity of statocytes and on the graviperception of cress roots,” Planta 157, 64–70 (1983).
[CrossRef] [PubMed]

Hiraoka, Y.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

Ketelaar, T.

T. Ketelaar, C. Faivre-Moskalenko, J. J. Esseling, N. C. de Ruijter, C. S. Grierson, M. Dogterom, A. M. Emons, “Positioning of nuclei in Arabidopsis root hairs: an actin-regulated process of tip growth,” Plant Cell 14, 2941–2955 (2002).

Landick, R.

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

Liang, H.

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

Liou, G. F.

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

Maiti, S.

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

Marion, S.

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

McCully, E. K.

E. K. McCully, C. F. Robinow, “Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy,” J. Cell Sci. 9, 475–507 (1971).
[PubMed]

Miller, M. J.

M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
[CrossRef] [PubMed]

Munteanu, E.-L.

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

Murray, A.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Nabeshima, K.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Nakagawa, T.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Neuman, K. C.

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

Nikitin, A. Y.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

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F. Chang, P. Nurse, “How fission yeast fission in the middle,” Cell 84, 191–194 (1996).
[CrossRef] [PubMed]

Oddershede, L.

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

Parker, I.

M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
[CrossRef] [PubMed]

Pavone, F. S.

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

L. Sacconi, E. Froner, R. Antolini, M. R. Taghizadeh, A. Choudhury, F. S. Pavone, “Multiphoton multifocal microscopy exploiting a diffractive optical element,” Opt. Lett. 28, 1918–1920 (2003).
[CrossRef] [PubMed]

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”

Peterman, E. J.

E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).

Profeta, G. A.

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Reinsch, S.

S. Reinsch, P. Gonczy, “Mechanisms of nuclear positioning,” J. Cell Sci. 111, 2283–2295 (1998).
[PubMed]

Rieder, C. L.

S. P. Alexander, C. L. Rieder, “Chromosome motion during attachment to the vertebrate spindle: initial saltatory-like behavior of chromosomes and quantitative analysis of force production by nascent kinetochore fibers,” J. Cell Biol. 113, 805–815 (1991).

Robinow, C. F.

E. K. McCully, C. F. Robinow, “Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy,” J. Cell Sci. 9, 475–507 (1971).
[PubMed]

Romano, G.

M. Capitanio, G. Romano, R. Ballerini, D. Dunlap, L. Finzi, M. Giuntini, F. S. Pavone, “Calibration of an optical tweezers with DIC signal,” Rev. Sci. Instrum. 73, 1687–1696 (2002).
[CrossRef]

Sacconi, L.

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

L. Sacconi, E. Froner, R. Antolini, M. R. Taghizadeh, A. Choudhury, F. S. Pavone, “Multiphoton multifocal microscopy exploiting a diffractive optical element,” Opt. Lett. 28, 1918–1920 (2003).
[CrossRef] [PubMed]

I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”

Schmidt, C. F.

E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).

K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
[CrossRef] [PubMed]

Schnapp, B. J.

K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
[CrossRef] [PubMed]

Shear, J. B.

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

Sievers, A.

A. Sievers, L. Heyder-Caspers, “The effect of centrifugal accelerations on the polarity of statocytes and on the graviperception of cress roots,” Planta 157, 64–70 (1983).
[CrossRef] [PubMed]

Straight, A. F.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Strickler, J. H.

W. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Stringari, C.

I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”

Svoboda, K.

K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, “Direct observation of kinesin stepping by optical trapping interferometry,” Nature 365, 721–727 (1993).
[CrossRef] [PubMed]

Taghizadeh, M. R.

Thon, G.

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

Tolic-Norrelykke, I. M.

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”

Tolic-Nørrelykke, I. M.

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

I. M. Tolic-Nørrelykke, E.-L. Munteanu, G. Thon, L. Oddershede, K. Berg-Sørensen, “Anomalous diffusion in living yeast cells,” Phys. Rev. Lett. 90, 078102 (2004).
[CrossRef]

Tomita, Y.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

Tromberg, B. J.

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Voigt, H.

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

Walter, R. J.

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Wang, M. D.

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

Webb, W. W.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Bio-technol. 21, 1369–1377 (2003).
[CrossRef]

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Wei, S. H.

M. J. Miller, S. H. Wei, I. Parker, M. D. Cahalan, “Two-photon imaging of lymphocyte motility and antigen response in intact lymph node,” Science 296, 1869–1873 (2002).
[CrossRef] [PubMed]

Wilhelm, C.

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

Williams, R. M.

W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Bio-technol. 21, 1369–1377 (2003).
[CrossRef]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

Wilson, K. L.

Y. Gruenbaum, K. L. Wilson, A. Harel, M. Goldberg, M. Cohen, “Review: nuclear lamins—structural proteins with fundamental functions,” J. Struct. Biol. 129, 313–323 (2000).
[CrossRef] [PubMed]

Wright, W. H.

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

M. W. Berns, W. H. Wright, B. J. Tromberg, G. A. Profeta, J. J. Andrews, R. J. Walter, “Use of a laser-induced optical force trap to study chromosome movement on the mitotic spindle,” Proc. Natl. Acad. Sci. U.S.A. 86, 4539–4543 (1989).

Yamamoto, A.

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

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A. Ashkin, J. M. Dziedzic, T. Yamane, “Optical trapping and manipulation of single cells using infrared laser beams,” Nature 330, 769–771 (1987).
[CrossRef] [PubMed]

Yamashita, Y. M.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Yanagida, M.

K. Nabeshima, T. Nakagawa, A. F. Straight, A. Murray, Y. Chikashige, Y. M. Yamashita, Y. Hiraoka, M. Yanagida, “Dynamics of centromeres during metaphase-anaphase transition in fission yeast: Dis 1 is implicated in force balance in metaphase bipolar spindle,” Mol. Biol. Cell 9, 3211–3225 (1998).

Yin, H.

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100, 7075–7080 (2003).

W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Bio-technol. 21, 1369–1377 (2003).
[CrossRef]

S. Maiti, J. B. Shear, R. M. Williams, W. R. Zipfel, W. W. Webb, “Measuring serotonin distribution in live cells with three-photon excitation,” Science 275, 530–532 (1997).
[CrossRef] [PubMed]

Biophys. J. (3)

K. C. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, S. M. Block, “Characterization of photodamage to escherichia coli in optical traps,” Biophys. J. 77, 2856–2863 (1999).

E. J. Peterman, F. Gittes, C. F. Schmidt, “Laser-induced heating in optical traps,” Biophys. J. 84, 1308–1316 (2003).

M. D. Wang, H. Yin, R. Landick, J. Gelles, S. M. Block, “Stretching DNA with optical tweezers,” Biophys. J. 72, 1335–1346 (1997).

Cell (1)

F. Chang, P. Nurse, “How fission yeast fission in the middle,” Cell 84, 191–194 (1996).
[CrossRef] [PubMed]

Curr Biol (1)

I. M. Tolic-Norrelykke, L. Sacconi, G. Thon, F. S. Pavone, “Positioning and elongation of the fission yeast spindle by microtubule-based pushing,” Curr Biol 14, 1181–1186 (2004).
[CrossRef] [PubMed]

Curr. Biol. (1)

D. R. Drummond, R. A. Cross, “Dynamics of interphase microtubules in Schizosaccharomyces pombe,” Curr. Biol. 10, 766–775 (2000).
[CrossRef] [PubMed]

Exp. Cell Res. (1)

M. W. Berns, J. R. Aist, W. H. Wright, H. Liang, “Optical trapping in animal and fungal cells using a tunable, near-infrared titanium-sapphire laser,” Exp. Cell Res. 198, 375–378 (1992).
[CrossRef] [PubMed]

Genes Cells (1)

D. Q. Ding, Y. Tomita, A. Yamamoto, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library,” Genes Cells 5, 169–190 (2000).
[CrossRef] [PubMed]

IEEE J. Sel. Top. Quantum Electron. (1)

A. Ashkin, “History of optical trapping and manipulation of small-neutral particle, atoms, and molecules,” IEEE J. Sel. Top. Quantum Electron. 6, 841–856 (2000).
[CrossRef]

J. Biomed. Opt. (1)

L. Sacconi, I. M. Tolic-Nørrelykke, R. Antolini, F. S. Pavone, “Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope,” J. Biomed. Opt. 10, 014002 (2005).
[CrossRef]

J. Cell Biol. (1)

S. P. Alexander, C. L. Rieder, “Chromosome motion during attachment to the vertebrate spindle: initial saltatory-like behavior of chromosomes and quantitative analysis of force production by nascent kinetochore fibers,” J. Cell Biol. 113, 805–815 (1991).

J. Cell Sci. (4)

S. Marion, C. Wilhelm, H. Voigt, J. C. Bacri, N. Guillen, “Overexpression of myosin IB in living Entamoeba histolytica enhances cytoplasm viscosity and reduces phagocytosis,” J. Cell Sci. 117, 3271–3279 (2004).
[CrossRef] [PubMed]

E. K. McCully, C. F. Robinow, “Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy,” J. Cell Sci. 9, 475–507 (1971).
[PubMed]

D. Q. Ding, Y. Chikashige, T. Haraguchi, Y. Hiraoka, “Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells,” J. Cell Sci. 111, 701–712 (1998).
[PubMed]

S. Reinsch, P. Gonczy, “Mechanisms of nuclear positioning,” J. Cell Sci. 111, 2283–2295 (1998).
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Other (1)

I. M. Tolic-Norrelykke, L. Sacconi, C. Stringari, F. S. Pavone are preparing a manuscript to be called “Nuclear and division plane positioning studied with optical tweezers.”

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Figures (6)

Fig. 1
Fig. 1

Experimental setup.

Fig. 2
Fig. 2

Trapping of a lipid granule. A sequence of bright-field images of a fission yeast cell, in which a lipid granule (arrow) is optically trapped and moved through the cytoplasm.

Fig. 3
Fig. 3

(A) Cells are alive, growing, and dividing after laser trapping. Two sister cells were chosen (inset); a granule was laser trapped for 5 min in the cell marked L+, whereas the cell marked L– was intact. Cell length is shown as a function of time. Both cells grew and divided (marked with an asterisk), but the cell with trapped granules showed a delay of ∼7 h with respect to the intact cell. This delay indicates a moderate laser-induced stress, which the cell can manage. (B) Microtubules are not significantly affected by trapping. A three-dimensional rendering of the interphase microtubule array before (1) and (2) after laser trapping. (C) Trapping does not perturb progression through mitosis. Selected images of a cell in anaphase B without laser trapping (L–) and with trapping (L+). The images marked 1 were taken in midanaphase B, whereas the images marked 2 were taken 7 min later, at the end of anaphase B. In the L+ cell, a granule was trapped close to the cell center between the acquisition of the two images for 5 min. Note that the behavior of the two spindles is very similar. (D) and (E) Trapping does not induce a detectable heating of the cell. (D) Spindle elongation rate (mean ± s.t.d.) in cells in which a granule was trapped for 5 min was not significantly different from that in cells without laser trapping. (E) Spindle elongation rate as a function of temperature. Measurements (circles) are taken from Refs. 28 (data at 20 °C and 36 °C) and 29 (data at 24.5 °C). The shaded area shows our measurements of the spindle elongation rate in cells with laser trapping at room temperature (horizontal lines, mean ± s.t.d.) and the corresponding temperature range (vertical lines). These measurements were performed at room temperature (21 °C–22 °C) and correspond to a range of 17.5 °C–21.5 °C on the graph, suggesting that the cell was not substantially heated by the laser trap. Scale bar in (B) and (C), 2 μm.

Fig. 4
Fig. 4

Scheme of the scanning-trap procedure for displacing the cell nucleus. A lipid granule (small sphere) is optically trapped close to the nucleus (large sphere). The granule is moved along a raster-scanning trajectory in the yz plane (small arrow), and simultaneously in the x direction to push the nucleus (large arrow).

Fig. 5
Fig. 5

Optically induced displacement of the nucleus. (A) Left, a singe optical section of the cell before the optical manipulation. The nucleus is in the cell center. Middle, the cell after the manipulation. Right, superposition of the two images, showing the displacement of the nucleus. Scale bar, 2 μm. (B) Set of optical z sections of the cell before (1) and after (2) the manipulation, showing that the whole volume of the nucleus was displaced.

Fig. 6
Fig. 6

Nucleus moves spontaneously back to the cell center after the optical manipulation. (A) Sequence of images of the cell before manipulation (the first image) and after the manipulation (the rest of the sequence). Numbers refer to time in minutes. The red lines serve as guides to the eye. Scale bar, 2 μm. Bottom, nuclear position as a function of time. The nucleus was displaced by 1.2 μm, after which it returned toward the cell center. The error on each data point is ∼50 nm.

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