Abstract

We describe the promising development of third-harmonic generation (THG) in laser scanning microscopy for study of the functional imaging of live biological cells. The dynamics of Ca2+ in biological cells is shown. The Ca2+ signal consists of a transient increase in the intracellular concentration. THG microscopy allows one to temporally visualize the release of Ca2+ from internal stores and (or) calcium influx.

© 2001 Optical Society of America

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  1. T. Y. F. Tsang, Phys. Rev. A 52, 4116 (1995).
    [CrossRef] [PubMed]
  2. G. C. Bjorklund, IEEE J. Quantum Electron. 11, 287 (1975).
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    [CrossRef] [PubMed]
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    [CrossRef]
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    [CrossRef] [PubMed]
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    [PubMed]
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    [CrossRef]

1999 (4)

1998 (1)

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, J. Microsc. 191, 266 (1998).
[CrossRef]

1997 (1)

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

1995 (2)

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

T. Y. F. Tsang, Phys. Rev. A 52, 4116 (1995).
[CrossRef] [PubMed]

1994 (1)

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

1989 (1)

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

1975 (1)

G. C. Bjorklund, IEEE J. Quantum Electron. 11, 287 (1975).
[CrossRef]

Audin, J.

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

Audy, M. C.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Barad, Y.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Bjorklund, G. C.

G. C. Bjorklund, IEEE J. Quantum Electron. 11, 287 (1975).
[CrossRef]

Brakenhoff, G. J.

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, J. Microsc. 191, 266 (1998).
[CrossRef]

Cantereau, A.

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

Chouï, M. T. V.

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

Djiane, J.

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

Dufy, B.

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

Dufy, B. J.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Eisenberg, H.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Fittinghoff, D. N.

Georgescauld, D.

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

Horowitz, M.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Hugues, A. R.

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

Millard, A. C.

Müller, A.

Müller, M.

Odessa, M. F.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Paly, J.

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

Perret, S.

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

Prevarskaya, N.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Putney, J. W.

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

Silberberg, Y.

D. Yelin and Y. Silberberg, Opt. Express 5, 169 (1999), http://epubs.osa.org/opticsexpress .
[CrossRef] [PubMed]

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Skryma, R.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Squier, J.

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, J. Microsc. 191, 266 (1998).
[CrossRef]

Squier, J. A.

Takemura, H.

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

Thastrup, O.

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

Tsang, T. Y. F.

T. Y. F. Tsang, Phys. Rev. A 52, 4116 (1995).
[CrossRef] [PubMed]

Vacher, A. M.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Vacher, P.

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

Wilson, K. R.

Wiseman, P. W.

Yelin, D.

Appl. Opt. (2)

Appl. Phys. Lett. (1)

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Biomed. Sci. (1)

P. Vacher, N. Prevarskaya, R. Skryma, M. C. Audy, A. M. Vacher, M. F. Odessa, and B. J. Dufy, Biomed. Sci. 2, 357 (1995).
[CrossRef]

Cell Calcium (1)

S. Perret, A. Cantereau, J. Audin, B. Dufy, and D. Georgescauld, Cell Calcium 25, 297 (1999).
[CrossRef] [PubMed]

Endocrinology (1)

P. Vacher, M. T. V. Chouï, J. Paly, J. Djiane, and B. Dufy, Endocrinology 134, 1213 (1994).
[PubMed]

IEEE J. Quantum Electron. (1)

G. C. Bjorklund, IEEE J. Quantum Electron. 11, 287 (1975).
[CrossRef]

J. Biol. Chem. (1)

H. Takemura, A. R. Hugues, O. Thastrup, and J. W. Putney, J. Biol. Chem. 264, 12,266 (1989).

J. Microsc. (1)

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, J. Microsc. 191, 266 (1998).
[CrossRef]

Opt. Express (1)

Phys. Rev. A (1)

T. Y. F. Tsang, Phys. Rev. A 52, 4116 (1995).
[CrossRef] [PubMed]

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Figures (3)

Fig. 1
Fig. 1

Schematic of the laser scanning THG microscope. The product used to change the intracellular Ca2+ is injected with a micropipette into a Petri box filled with a saline solution.

Fig. 2
Fig. 2

THG microscopy images of one part of a fusiform glial cell. (a) Reference image. Note that the image contrast is due to THG intensity, principally at the interfaces. (b) Image just after injection of thapsigargin. (c), (d) Images of the same part of the glial cell at t0+6 mn and t0+12 mn, respectively.

Fig. 3
Fig. 3

Mean THG intensity value measured at the two square surfaces of the cell shown in Fig.  2(a) as a function of time. Circles and triangles represent the right and left square surfaces, respectively.

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