Abstract

The structural origin of scattering contrast from single cells is examined by using a combined optical coherence and multiphoton microscope based on a 12fs Ti:sapphire source and a 0.95 NA objective. High-resolution coherence-gated scattering images from single cells are coregistered and compared with two-photon-excited fluorescence images. Scattering contrast is observed from mitochondria, plasma membrane, actin filaments, and the boundary between cytoplasm and nucleus. There is little contribution to scattering from regions inside the nuclear core. These results confirm that light scattering signals from specific subcellular structures can be visualized by using coherent reflectance geometry.

© 2007 Optical Society of America

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C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

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R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

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J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

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[CrossRef] [PubMed]

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[CrossRef] [PubMed]

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[CrossRef] [PubMed]

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[CrossRef] [PubMed]

Boccara, C.

Boiko, L.

R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

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[CrossRef]

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B. Beauvoit, T. Kitai, and B. Chance, Biophys. J. 67, 2501 (1994).
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S. Tang, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, J. Biomed. Opt. 11, 020502 (2006).
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R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
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R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
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J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
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S. A. Boppart, B. E. Bouma, C. Pitris, J. F. Southern, M. E. Brezinski, and J. G. Fujimoto, Nat. Med. 4, 861 (1998).
[CrossRef] [PubMed]

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Guerra, A.

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[CrossRef] [PubMed]

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R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

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[CrossRef] [PubMed]

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B. Beauvoit, T. Kitai, and B. Chance, Biophys. J. 67, 2501 (1994).
[CrossRef] [PubMed]

Krasieva, T. B.

S. Tang, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Lecaque, R.

Luo, W.

C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

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R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

Malpica, A.

R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

Marks, D. L.

C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

Mertz, J.

Moneron, G.

Moreaux, L.

Mourant, J. R.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Pitris, C.

S. A. Boppart, B. E. Bouma, C. Pitris, J. F. Southern, M. E. Brezinski, and J. G. Fujimoto, Nat. Med. 4, 861 (1998).
[CrossRef] [PubMed]

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C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

Richards-Kortum, R.

R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

Southern, J. F.

S. A. Boppart, B. E. Bouma, C. Pitris, J. F. Southern, M. E. Brezinski, and J. G. Fujimoto, Nat. Med. 4, 861 (1998).
[CrossRef] [PubMed]

Tan, W.

C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

Tang, S.

S. Tang, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Tromberg, B. J.

S. Tang, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Vabre, L.

Vinegoni, C.

C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

Appl. Opt.

Appl. Phys. Lett.

C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
[CrossRef]

Biophys. J.

B. Beauvoit, T. Kitai, and B. Chance, Biophys. J. 67, 2501 (1994).
[CrossRef] [PubMed]

J. Biomed. Opt.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

R. Drezek, M. Guillaud, T. Collier, L. Boiko, A. Malpica, C. Macaulay, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 8, 7 (2003).
[CrossRef] [PubMed]

S. Tang, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Nat. Med.

S. A. Boppart, B. E. Bouma, C. Pitris, J. F. Southern, M. E. Brezinski, and J. G. Fujimoto, Nat. Med. 4, 861 (1998).
[CrossRef] [PubMed]

Opt. Lett.

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Figures (2)

Fig. 1
Fig. 1

Comparison of TPEF (left), OCT (middle), and merged (right) images of single cells. (a)–(c) Single cell without labeling; (d)–(f) with DAPI labeling for nuclei; (g)–(i) with Rhodamine 123 labeling for mitochondria; (j)–(l) with PKH67 labeling for plasma membrane; (m)–(o) with Alexa Fluor 488 conjugated to phalloidin for actin filaments. In the merged images, TPEF and OCT signals are in green and red, respectively. The scale bar is 10 μ m .

Fig. 2
Fig. 2

Multiple frames of OCT images of a single cell optically sliced from the top to the bottom. From (a) to (k), the images are taken at progressive 1 μ m Z steps. (l) is the X Y projection of the whole Z stack of cell slices. The scale bar is 10 μ m .

Tables (1)

Tables Icon

Table 1 Normalized Mean Scattering Intensity and Scattering Area by Region a

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