Abstract

We demonstrate a novel multifocal, multiphoton microscope that is capable of simultaneous dynamic imaging of multiple focal planes. We show for the first time that multimodal, multiphoton images excited with orthogonal polarizations can be acquired simultaneously in both the transmission and epi directions.

© 2008 Optical Society of America

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  1. P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
    [CrossRef]
  2. B. Rózsa, G. Katona, E. S. Vizi, Z. Várallyay, A. Sághy, L. Valenta, P. Maák, J. Fekete, A. Bányász, and R. Szipöcs, "Random access three-dimensional two-photon microscopy," Appl. Opt. 46, 1860-1865 (2007).
    [CrossRef]
  3. W. Amir, R. Carriles, E. E. Hoover, T. A. Planchon, C. G. Durfee, and J. A. Squier, "Simultaneous imaging of multiple focal planes using a two-photon scanning microscope," Opt. Lett. 32, 1731-1733 (2007).
    [CrossRef]
  4. J. O. Hamblen, T. S. Hall, and M. D. Furman, Rapid Prototyping of Digital Systems: Quartus II Edition (Springer, 2006).
  5. R. Carriles, E. E. Hoover, W. Amir, and J. A. Squier, "Simultaneous imaging of multiple focal planes in scanning two-photon absorption microscope by photon counting," Proc. SPIE 6771, 67710C-2 - 67710C-8 (2007).
  6. J. Bewersdorf, R. Pick, and S. W. Hell, "Multifocal multiphoton microscopy," Opt. Lett. 23, 655-657 (1998).
    [CrossRef]
  7. A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
    [CrossRef]
  8. M. Fricke, and T. Nielsen, "Two-dimensional imaging without scanning by multifocal multiphoton microscopy," Appl. Opt. 44, 2984-2988 (2005).
    [CrossRef]
  9. D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
    [CrossRef]
  10. G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).
  11. S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr:forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
    [CrossRef]
  12. K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
    [CrossRef]
  13. R. Cisek, N. Prent, C. Greenhalgh, D. Sandkuijl, A. Tuer, A. Major, and V. Barzda, "Multicontrast nonlinear imaging microscopy," in Biochemical Applications of Nonlinear Optical Spectroscopy, V. V. Yakovlev, ed. (CRC Press, in press).

2008 (1)

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

2007 (2)

2005 (1)

2004 (1)

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

2001 (1)

2000 (1)

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

1998 (2)

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

J. Bewersdorf, R. Pick, and S. W. Hell, "Multifocal multiphoton microscopy," Opt. Lett. 23, 655-657 (1998).
[CrossRef]

1997 (1)

D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
[CrossRef]

Amat-Roldán, I.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Amir, W.

Artigas, D.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Baldwin, P. M.

D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
[CrossRef]

Bányász, A.

Bewersdorf, J.

Bouchet, B.

D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
[CrossRef]

Brakenhoff, G. J.

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

Buist, A. H.

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

Carriles, R.

Chen, I. H.

Chen, P. C.

Chu, S. W.

Cormack, I. G.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Durfee, C. G.

Fekete, J.

Fricke, M.

Gallant, D. J.

D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
[CrossRef]

Gualda, E. J.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Hell, S. W.

Hoover, E. E.

Katona, G.

Lin, B. L.

Liu, T. M.

Loza-Alvarez, P.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Maák, P.

Mathew, M.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Mizutani, G.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Müller, M.

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

Nielsen, T.

Ober, R. J.

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

Pick, R.

Planchon, T. A.

Prabhat, P.

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

Psilodimitrakopoulos, S.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Ram, S.

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

Rózsa, B.

Sághy, A.

Sakamoto, M.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Sano, H.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Sonoda, Y.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Squier, J.

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

Squier, J. A.

Sun, C. K.

Szipöcs, R.

Takahashi, T.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Thayil, K. N. A.

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

Ushioda, S.

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

Valenta, L.

Várallyay, Z.

Vizi, E. S.

Ward, E. S.

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

Appl. Opt. (2)

Carbohydr. Polym. (1)

D. J. Gallant, B. Bouchet, and P. M. Baldwin, "Microscopy of starch: evidence of a new level of granule organization," Carbohydr. Polym. 32, 177-191 (1997).
[CrossRef]

IEEE Trans. Nanobiosci. (1)

P. Prabhat, S. Ram, E. S. Ward, and R. J. Ober, "Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions," IEEE Trans. Nanobiosci. 3, 237-242 (2004).
[CrossRef]

J. Lumin. (1)

G. Mizutani, Y. Sonoda, H. Sano, M. Sakamoto, T. Takahashi, and S. Ushioda, "Detection of starch granules in a living plant by optical second harmonic microscopy," J. Lumin. 87-89, 824-826 (2000).

J. Microsc. (2)

K. N. A. Thayil, E. J. Gualda, S. Psilodimitrakopoulos, I. G. Cormack, I. Amat-Roldán, M. Mathew, D. Artigas, and P. Loza-Alvarez, "Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy," J. Microsc. 230, 70-75 (2008).
[CrossRef]

A. H. Buist, M. Müller, J. Squier, and G. J. Brakenhoff, "Real time two-photon absorption microscopy using multi point excitation," J. Microsc. 192, 217-226 (1998).
[CrossRef]

Opt. Lett. (3)

Other (3)

J. O. Hamblen, T. S. Hall, and M. D. Furman, Rapid Prototyping of Digital Systems: Quartus II Edition (Springer, 2006).

R. Carriles, E. E. Hoover, W. Amir, and J. A. Squier, "Simultaneous imaging of multiple focal planes in scanning two-photon absorption microscope by photon counting," Proc. SPIE 6771, 67710C-2 - 67710C-8 (2007).

R. Cisek, N. Prent, C. Greenhalgh, D. Sandkuijl, A. Tuer, A. Major, and V. Barzda, "Multicontrast nonlinear imaging microscopy," in Biochemical Applications of Nonlinear Optical Spectroscopy, V. V. Yakovlev, ed. (CRC Press, in press).

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