Abstract

Combining the noninterferometric wide-field optical profilometry technique with fluorescence microscopy, we observe the membrane activities of a living cell as well as the structures of its cytoskeletons. The membrane ripples of a lamellipodium are related to similar structures of the underlying actin filaments. However, we find the ripples appear prior to and disappear later than the corresponding actin filament structures, which supports the elastic Brownian ratchet model of cell motility. In addition, we measure the three-dimensional movement of a fibronectin-coated latex bead on the membrane. The bead motion is determined by the movement and branching of the actin molecules on the filament, as well as by the displacement of the filament itself.

© 2006 Optical Society of America

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  1. T. D. Pollard and G. G. Borisy, Cell 112, 453 (2003).
    [CrossRef] [PubMed]
  2. G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
    [CrossRef]
  3. K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
    [CrossRef] [PubMed]
  4. S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
    [CrossRef] [PubMed]
  5. A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
    [CrossRef] [PubMed]
  6. J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
    [CrossRef]
  7. M. A. A. Neil, R. Juskaitis, and T. Wilson, Opt. Lett. 22, 1905 (1997).
    [CrossRef]
  8. C.-H. Lee and J. Wang, Opt. Commun. 135, 233 (1997).
    [CrossRef]
  9. C.-H. Lee, H.-Y. Mong, and W.-C. Lin, Opt. Lett. 27, 1773 (2002).
    [CrossRef]
  10. C.-C. Wang, J.-Y. Lin, and C.-H. Lee, Opt. Express 13, 10665 (2005).
    [CrossRef] [PubMed]
  11. B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).
  12. H. T. McMahon and J. L. Gallop, Nature 438, 590 (2005).
    [CrossRef] [PubMed]
  13. A. Mogilner and G. Oster, Biophys. J. 71, 3030 (1996).
    [CrossRef] [PubMed]
  14. N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
    [CrossRef] [PubMed]
  15. T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
    [CrossRef] [PubMed]
  16. S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
    [CrossRef] [PubMed]

2006 (1)

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

2005 (5)

H. T. McMahon and J. L. Gallop, Nature 438, 590 (2005).
[CrossRef] [PubMed]

S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
[CrossRef] [PubMed]

A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
[CrossRef] [PubMed]

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

C.-C. Wang, J.-Y. Lin, and C.-H. Lee, Opt. Express 13, 10665 (2005).
[CrossRef] [PubMed]

2003 (2)

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

T. D. Pollard and G. G. Borisy, Cell 112, 453 (2003).
[CrossRef] [PubMed]

2002 (1)

2001 (1)

G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
[CrossRef]

2000 (1)

T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
[CrossRef] [PubMed]

1997 (2)

1996 (1)

A. Mogilner and G. Oster, Biophys. J. 71, 3030 (1996).
[CrossRef] [PubMed]

1993 (1)

N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
[CrossRef] [PubMed]

Alberts, B.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Bittman, R.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Blanchoin, L.

T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
[CrossRef] [PubMed]

Borisy, G. G.

T. D. Pollard and G. G. Borisy, Cell 112, 453 (2003).
[CrossRef] [PubMed]

Buranday, T.

A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
[CrossRef] [PubMed]

Burns, A. R.

A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
[CrossRef] [PubMed]

Butler, J. P.

N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
[CrossRef] [PubMed]

Cardenas, R.

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

Discher, D. E.

S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
[CrossRef] [PubMed]

Epand, R. F.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Epand, R. M.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Frankel, D. J.

A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
[CrossRef] [PubMed]

Gallop, J. L.

H. T. McMahon and J. L. Gallop, Nature 438, 590 (2005).
[CrossRef] [PubMed]

Gaus, K.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Gelissen, I.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Gratton, E.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Ingber, D. E.

N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
[CrossRef] [PubMed]

Jessup, W.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Johnson, A.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Jones, A. S.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Juskaitis, R.

Kable, E. P. W.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Kas, J.

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

Koch, D.

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

Kritharides, L.

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Lee, C.-H.

Lewis, J.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Li, Z.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Lin, J.-Y.

Lin, W.-C.

McMahon, H. T.

H. T. McMahon and J. L. Gallop, Nature 438, 590 (2005).
[CrossRef] [PubMed]

Mogilner, A.

A. Mogilner and G. Oster, Biophys. J. 71, 3030 (1996).
[CrossRef] [PubMed]

Mong, H.-Y.

Mullins, R. D.

T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
[CrossRef] [PubMed]

Neil, M. A. A.

Oster, G.

A. Mogilner and G. Oster, Biophys. J. 71, 3030 (1996).
[CrossRef] [PubMed]

Park, S.

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

Pollard, T. D.

T. D. Pollard and G. G. Borisy, Cell 112, 453 (2003).
[CrossRef] [PubMed]

T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
[CrossRef] [PubMed]

Raff, M.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Roberts, K.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Schindler, H.

G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
[CrossRef]

Schutz, G. J.

G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
[CrossRef]

Sen, S.

S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
[CrossRef] [PubMed]

Shaw, J. E.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Shih, C. K.

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

Sonnleitner, M.

G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
[CrossRef]

Subramanian, S.

S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
[CrossRef] [PubMed]

Walter, P.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

Wang, C.-C.

Wang, J.

C.-H. Lee and J. Wang, Opt. Commun. 135, 233 (1997).
[CrossRef]

Wang, N.

N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
[CrossRef] [PubMed]

Wilson, T.

Yip, C. M.

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

Annu. Rev. Biophys. Biomol. Struct. (1)

T. D. Pollard, L. Blanchoin, and R. D. Mullins, Annu. Rev. Biophys. Biomol. Struct. 29, 545 (2000).
[CrossRef] [PubMed]

Biophys. J. (5)

S. Park, D. Koch, R. Cardenas, J. Kas, and C. K. Shih, Biophys. J. 89, 4330 (2005).
[CrossRef] [PubMed]

S. Sen, S. Subramanian, and D. E. Discher, Biophys. J. 89, 3203 (2005).
[CrossRef] [PubMed]

A. R. Burns, D. J. Frankel, and T. Buranday, Biophys. J. 89, 1081 (2005).
[CrossRef] [PubMed]

J. E. Shaw, R. F. Epand, R. M. Epand, Z. Li, R. Bittman, and C. M. Yip, Biophys. J. 90, 2170 (2006).
[CrossRef]

A. Mogilner and G. Oster, Biophys. J. 71, 3030 (1996).
[CrossRef] [PubMed]

Cell (1)

T. D. Pollard and G. G. Borisy, Cell 112, 453 (2003).
[CrossRef] [PubMed]

J. Fluoresc. (1)

G. J. Schutz, M. Sonnleitner, and H. Schindler, J. Fluoresc. 11, 177 (2001).
[CrossRef]

Nature (1)

H. T. McMahon and J. L. Gallop, Nature 438, 590 (2005).
[CrossRef] [PubMed]

Opt. Commun. (1)

C.-H. Lee and J. Wang, Opt. Commun. 135, 233 (1997).
[CrossRef]

Opt. Express (1)

Opt. Lett. (2)

Proc. Natl. Acad. Sci. U.S.A. (1)

K. Gaus, E. Gratton, E. P. W. Kable, A. S. Jones, I. Gelissen, L. Kritharides, and W. Jessup, Proc. Natl. Acad. Sci. U.S.A. 100, 15554 (2003).
[CrossRef] [PubMed]

Science (1)

N. Wang, J. P. Butler, and D. E. Ingber, Science 260, 1124 (1993).
[CrossRef] [PubMed]

Other (1)

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Molecular Biology of the Cell, 4th ed. (Garland Science, 2002).

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Figures (4)

Fig. 1
Fig. 1

Images of a fibroblast (a) bright-field reflection image, (b) NIWOP topography, (c) GFP–actin image. In (b) and (c) the region enclosed by a dashed rectangle is the lamellipodium analyzed in Figs. 2, 3.

Fig. 2
Fig. 2

Time-lapse images of the membrane motion and the cortex actin of a lamellipodium. In each panel the top image is the membrane topography obtained by NIWOP and the bottom image is the actin distribution. Concentric membrane ripples appear in (a) and then propagate from left to right. However, concentric structures of the actin filaments do not appear until 30 min (b). After 210 min , the membrane ripples spread over this lamellipodium (d).

Fig. 3
Fig. 3

Time-lapse images of the membrane motion and the cortex actin after treatment with 20 μ M cytochalasin D. Each panel indicates the length of time after the treatment. The concentric filament structure is dissolved after 30 min (b). However, the membrane ripples remain for 40 min (c). The membrane topography then becomes random, with many spikes on the leading edge.

Fig. 4
Fig. 4

Motions of a 1 μ m fibronectin-coated bead on the membrane and the underlying actin filament. The dashed lines represent the cell edges. The bead, marked by a white triangle, is bound to an actin filament. The brighter fluorescence spot on the filament signifies aggregation of actin molecules on the same position. The arrows in (a) indicate the directions of movements of the bead and the filament.

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