Abstract

Because it avoids the in-focus photodamage and phototoxicity problem of two-photon-fluorescence excitation, multiharmonic-generation biopsy based on a 1200–1300-nm light source could provide a truly noninvasive and highly penetrative optical sectioning of skin. We study multiharmonic-generation biopsy of fixed mouse skin. Our preliminary study suggests that this technique could provide submicrometer-resolution deep-tissue noninvasive biopsy images in skin without the use of fluorescence and exogenous markers.

© 2003 Optical Society of America

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    [CrossRef] [PubMed]
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2002 (5)

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Y. Yasuno, S. Makita, Y. Sutoh, M. Itoh, and T. Yatagai, Opt. Lett. 27, 1803 (2002).
[CrossRef]

2001 (4)

B. R. Masters and P. T. C. So, Opt. Express 8, 2 (2001), http://www.opticsexpress.org.
[CrossRef] [PubMed]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

1999 (3)

1998 (1)

1997 (3)

1981 (1)

R. R. Anderson and J. A. Parish, J. Investig. Dermatol. 77, 13 (1981).
[CrossRef]

1978 (1)

H. Elias, J. E. Pauly, and E. R. Burns, Histology and Human Microanatomy, 4th ed. (Piccin, Padova, Italy, 1978).

Alfano, R. R.

Anderson, R. R.

R. R. Anderson and J. A. Parish, J. Investig. Dermatol. 77, 13 (1981).
[CrossRef]

Apolonski, A.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Bavister, B. D.

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, Nature Biotechnol. 17, 763 (1999).
[CrossRef]

Bizheva, K.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Boppart, S. A.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Bouma, B. E.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Brakenhoff, G. J.

Brezinski, M. E.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Burns, E. R.

H. Elias, J. E. Pauly, and E. R. Burns, Histology and Human Microanatomy, 4th ed. (Piccin, Padova, Italy, 1978).

Campagnola, P. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Chen, I.-H.

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

Chen, I.-S.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Chen, S.-Y.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Cheng, P. C.

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

Cheng, P.-C.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

Chu, S.-W.

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Drexler, W.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Elias, H.

H. Elias, J. E. Pauly, and E. R. Burns, Histology and Human Microanatomy, 4th ed. (Piccin, Padova, Italy, 1978).

Fercher, A. F.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Fujimoto, J. G.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Gratton, E.

Guo, Y.

Halbhuber, J.-J.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

Harris, D.

Hermann, B.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Ho, P. P.

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Itoh, M.

Johnson, I.

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

Kim, H.

Knight, J. C.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Kochevar, I. E.

König, K.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

K. König, P. T. C. So, W. W. Mantulin, and E. Gratton, Opt. Lett. 22, 135 (1997).
[CrossRef]

Krieg, R.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

Kuo, M.-X.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Lee, S.-P.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Lin, B.-L.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

Lin, C.-Y.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Lin, D.-J.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Liu, F.

Liu, H.-L.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

Liu, T.-M.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Makita, S.

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Mantulin, W. W.

Masters, B. R.

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Muller, M.

Parish, J. A.

R. R. Anderson and J. A. Parish, J. Investig. Dermatol. 77, 13 (1981).
[CrossRef]

Pauly, J. E.

H. Elias, J. E. Pauly, and E. R. Burns, Histology and Human Microanatomy, 4th ed. (Piccin, Padova, Italy, 1978).

Peuckert, C.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

Pitris, C.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Povazay, B.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Russell, P. St. J.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Sacks, P.

Sattmann, H.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Savage, H.

Schantz, S.

Scherzer, E.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Silberberg, Y.

So, P. T. C.

Southern, J. F.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Squier, J. A.

Squirell, J. M.

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, Nature Biotechnol. 17, 763 (1999).
[CrossRef]

Sun, C.-K.

I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

S.-W. Chu, I.-H. Chen, T.-M. Liu, P. C. Cheng, C.-K. Sun, and B.-L. Lin, Opt. Lett. 26, 1909 (2001).
[CrossRef]

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Sutoh, Y.

Tearney, G. J.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Tirlapur, U. K.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

Tsai, H.-J.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Tsai, T.-H.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

Unterhuber, A.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Vetterlein, M.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Wadsworth, W. J.

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

White, J. G.

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, Nature Biotechnol. 17, 763 (1999).
[CrossRef]

Wilson, K. R.

Wokosin, D. L.

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, Nature Biotechnol. 17, 763 (1999).
[CrossRef]

Yasuno, Y.

Yatagai, T.

Yelin, D.

Zhadin, N.

Biophys. J. (1)

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 81, 493 (2002).
[CrossRef]

Exp. Cell Res. (1)

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and J.-J. Halbhuber, Exp. Cell Res. 263, 88 (2001).
[CrossRef] [PubMed]

J. Investig. Dermatol. (1)

R. R. Anderson and J. A. Parish, J. Investig. Dermatol. 77, 13 (1981).
[CrossRef]

J. Microsc. (1)

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, J. Microsc. 208, 190 (2002).
[CrossRef] [PubMed]

J. Struct. Biol. (1)

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, “Harmonic generation microscopy for developmental biology,” J. Struct. Biol. (to be published).

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[CrossRef]

Opt. Lett. (1)

B. Povazay, K. Bizheva, A. Unterhuber, B. Hermann, H. Sattmann, A. F. Fercher, W. Drexler, A. Apolonski, W. J. Wadsworth, J. C. Knight, P. St. J. Russell, M. Vetterlein, and E. Scherzer, Opt. Lett. 27, 1800 (2002).
[CrossRef]

Opt. Express (4)

Opt. Lett. (4)

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I.-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, Opt. Quantum Electron. 34, 1251 (2002).
[CrossRef]

Scanning (1)

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, Scanning 23, 249 (2001).
[CrossRef] [PubMed]

Science (1)

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Figures (3)

Fig. 1
Fig. 1

(a) B-SHG, (b) F-SHG, (c) F-THG, and (d) the combined cross-sectional (xz, into the skin, top row) as well as the paradermal (xy, parallel to the surface, bottom row) scanning images of mouse skin. B-SHG, F-SHG, and F-THG signals are denoted by green, red, and blue, respectively. The dotted lines in the top (bottom) row mark the corresponding z (y) position for the bottom (top) row. Image size: 235 µm×310 µm for xz images and 235 µm×235 µm for xy images. i, cover glass–formalin interface; e, epidermis layer; de, dermis layer.

Fig. 2
Fig. 2

Paradermal multiharmonic-generation biopsy images of mouse skin. (a) Sectioned image in the stratum basale layer. Image size: 59 µm×59 µm. (b) Sectioned image of hairs above skin. (c) Sectioned image deep in the dermis. Image size for (b) and (c): 235 µm×235 µm. h, hair; s, sebaceous gland. B-SHG, F-SHG, and F-THG signals are denoted by green, red, and blue, respectively.

Fig. 3
Fig. 3

Multiharmonic-generation biopsy images of a mouse ear. (a) Paradermal sectioned F-SHG image inside the mouse’s external ear showing the elastic cartilage. Image size: 235 µm×235 µm. (b) and (c) are the 3D reconstructed transverse and longitudinal F-THG images of capillaries inside the mouse’s external ear, respectively. Image size: 80 µm×80 µm. p, capillary.

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