Abstract

Skin aging is an important issue in geriatric and cosmetic dermatology. To quantitatively analyze changes in keratinocytes related to intrinsic aging, we exploited a 1230 nm-based in vivo harmonic generation microscopy, combining second- and third-harmonic generation modalities. 52 individuals (21 men and 31 women, age range 19–79) were examined on the sun-protected volar forearm. Through quantitative analysis by the standard algorithm provided, we found that the cellular and nuclear size of basal keratinocytes, but not that of granular cells, was significantly increased with advancing age. The cellular and nuclear areas, which have an increase of 0.51 μm2 and 0.15 μm2 per year, respectively, can serve as scoring indices for intrinsic skin aging.

© 2012 OSA

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    [CrossRef] [PubMed]

2011 (4)

E. M. Buckingham and A. J. Klingelhutz, “The role of telomeres in the ageing of human skin,” Exp. Dermatol.20(4), 297–302 (2011).
[CrossRef] [PubMed]

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
[CrossRef] [PubMed]

M. R. Tsai, S. Y. Chen, D. B. Shieh, P. J. Lou, and C.-K. Sun, “In vivo optical virtual biopsy of human oral mucosa with harmonic generation microscopy,” Biomed. Opt. Express2(8), 2317–2328 (2011).
[CrossRef] [PubMed]

2010 (1)

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

2009 (1)

S. Y. Chen, H. Y. Wu, and C.-K. Sun, “In vivo harmonic generation biopsy of human skin,” J. Biomed. Opt.14(6), 060505 (2009).
[CrossRef] [PubMed]

2008 (5)

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
[CrossRef] [PubMed]

M. A. Farage, K. W. Miller, P. Elsner, and H. I. Maibach, “Intrinsic and extrinsic factors in skin ageing: a review,” Int. J. Cosmet. Sci.30(2), 87–95 (2008).
[CrossRef] [PubMed]

C. S. Hsieh, S. U. Chen, Y. W. Lee, Y. S. Yang, and C.-K. Sun, “Higher harmonic generation microscopy of in vitro cultured mammal oocytes and embryos,” Opt. Express16(15), 11574–11588 (2008).
[PubMed]

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

2007 (1)

P. J. Matts, P. J. Dykes, and R. Marks, “The distribution of melanin in skin determined in vivo,” Br. J. Dermatol.156(4), 620–628 (2007).
[CrossRef] [PubMed]

2006 (3)

M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
[CrossRef] [PubMed]

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

M. Sugimoto, R. Yamashita, and M. Ueda, “Telomere length of the skin in association with chronological aging and photoaging,” J. Dermatol. Sci.43(1), 43–47 (2006).
[CrossRef] [PubMed]

2005 (2)

J. M. Waller and H. I. Maibach, “Age and skin structure and function, a quantitative approach (I): blood flow, pH, thickness, and ultrasound echogenicity,” Skin Res. Technol.11(4), 221–235 (2005).
[CrossRef] [PubMed]

M.-C. Chan, T.-M. Liu, S.-P. Tai, and C.-K. Sun, “Compact fiber-delivered Cr:forsterite laser for nonlinear light microscopy,” J. Biomed. Opt.10(5), 054006 (2005).
[CrossRef] [PubMed]

2004 (1)

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
[CrossRef] [PubMed]

2003 (1)

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

2002 (3)

G. Jenkins, “Molecular mechanisms of skin ageing,” Mech. Ageing Dev.123(7), 801–810 (2002).
[CrossRef] [PubMed]

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

2001 (1)

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

1998 (3)

M. Gniadecka and G. B. Jemec, “Quantitative evaluation of chronological ageing and photoageing in vivo: studies on skin echogenicity and thickness,” Br. J. Dermatol.139(5), 815–821 (1998).
[CrossRef] [PubMed]

A. R. Haake, I. Roublevskaia, and M. Cooklis, “Apoptosis: a role in skin aging?” J. Investig. Dermatol. Symp. Proc.3(1), 28–35 (1998).
[PubMed]

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, “3D microscopy of transparent objects using third-harmonic generation,” J. Microsc.191(3), 266–274 (1998).
[CrossRef] [PubMed]

1995 (1)

T. Y. F. Tsang, “Optical third-harmonic generation at interfaces,” Phys. Rev. A52(5), 4116–4125 (1995).
[CrossRef] [PubMed]

1989 (1)

B. A. Gilchrest, “Skin aging and photoaging: an overview,” J. Am. Acad. Dermatol.21(3), 610–613 (1989).
[CrossRef] [PubMed]

1986 (1)

N. A. Fenske and C. W. Lober, “Structural and functional changes of normal aging skin,” J. Am. Acad. Dermatol.15(4), 571–585 (1986).
[CrossRef] [PubMed]

1985 (1)

Y. Barrandon and H. Green, “Cell size as a determinant of the clone-forming ability of human keratinocytes,” Proc. Natl. Acad. Sci. U.S.A.82(16), 5390–5394 (1985).
[CrossRef] [PubMed]

Ahmad, H.

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

Aoyagi, Y.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Arai, T.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Attia, S.

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

Barrandon, Y.

Y. Barrandon and H. Green, “Cell size as a determinant of the clone-forming ability of human keratinocytes,” Proc. Natl. Acad. Sci. U.S.A.82(16), 5390–5394 (1985).
[CrossRef] [PubMed]

Bauer, A.

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

Beaurepaire, E.

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Beretti, F.

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

Birk, D. E.

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

Bisschop, M. A.

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
[CrossRef] [PubMed]

Boucher, Y.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Brakenhoff, G. J.

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, “3D microscopy of transparent objects using third-harmonic generation,” J. Microsc.191(3), 266–274 (1998).
[CrossRef] [PubMed]

Brégégère, F. M.

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Brown, D.

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

Brown, E.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Buckingham, E. M.

E. M. Buckingham and A. J. Klingelhutz, “The role of telomeres in the ageing of human skin,” Exp. Dermatol.20(4), 297–302 (2011).
[CrossRef] [PubMed]

Bückle, R.

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
[CrossRef] [PubMed]

Casari, A.

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

Cesinaro, A. M.

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

Chan, M.-C.

M.-C. Chan, T.-M. Liu, S.-P. Tai, and C.-K. Sun, “Compact fiber-delivered Cr:forsterite laser for nonlinear light microscopy,” J. Biomed. Opt.10(5), 054006 (2005).
[CrossRef] [PubMed]

Chen, S. U.

Chen, S. Y.

Chen, S.-U.

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

Chen, S.-Y.

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

Cheng, P. C.

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

Chu, S.-W.

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

Combettes, L.

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Cooklis, M.

A. R. Haake, I. Roublevskaia, and M. Cooklis, “Apoptosis: a role in skin aging?” J. Investig. Dermatol. Symp. Proc.3(1), 28–35 (1998).
[PubMed]

Débarre, D.

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

diTomaso, E.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Dykes, P. J.

P. J. Matts, P. J. Dykes, and R. Marks, “The distribution of melanin in skin determined in vivo,” Br. J. Dermatol.156(4), 620–628 (2007).
[CrossRef] [PubMed]

El-Domyati, M.

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M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
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M. A. Farage, K. W. Miller, P. Elsner, and H. I. Maibach, “Intrinsic and extrinsic factors in skin ageing: a review,” Int. J. Cosmet. Sci.30(2), 87–95 (2008).
[CrossRef] [PubMed]

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
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M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
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D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
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M. A. Farage, K. W. Miller, P. Elsner, and H. I. Maibach, “Intrinsic and extrinsic factors in skin ageing: a review,” Int. J. Cosmet. Sci.30(2), 87–95 (2008).
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N. A. Fenske and C. W. Lober, “Structural and functional changes of normal aging skin,” J. Am. Acad. Dermatol.15(4), 571–585 (1986).
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M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
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M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
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A. R. Haake, I. Roublevskaia, and M. Cooklis, “Apoptosis: a role in skin aging?” J. Investig. Dermatol. Symp. Proc.3(1), 28–35 (1998).
[PubMed]

Hahn, S.

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
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Hsieh, C. S.

Izumiyama-Shimomura, N.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
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E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
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M. Gniadecka and G. B. Jemec, “Quantitative evaluation of chronological ageing and photoageing in vivo: studies on skin echogenicity and thickness,” Br. J. Dermatol.139(5), 815–821 (1998).
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T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

Kaatz, M.

M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
[CrossRef] [PubMed]

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
[CrossRef] [PubMed]

Kato, M.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Kawabata, K.

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
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K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Koehler, M. J.

M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
[CrossRef] [PubMed]

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
[CrossRef] [PubMed]

König, K.

M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
[CrossRef] [PubMed]

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett.31(19), 2879–2881 (2006).
[CrossRef] [PubMed]

Lee, W.-J.

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

Lee, Y. W.

Lenderink, E.

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
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Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Liao, Y.-H.

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

Lin, B.-L.

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

Liu, T.-M.

M.-C. Chan, T.-M. Liu, S.-P. Tai, and C.-K. Sun, “Compact fiber-delivered Cr:forsterite laser for nonlinear light microscopy,” J. Biomed. Opt.10(5), 054006 (2005).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
[CrossRef] [PubMed]

Lober, C. W.

N. A. Fenske and C. W. Lober, “Structural and functional changes of normal aging skin,” J. Am. Acad. Dermatol.15(4), 571–585 (1986).
[CrossRef] [PubMed]

Longo, C.

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

Lou, P. J.

Lucassen, G. W.

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
[CrossRef] [PubMed]

Ma’or, Z.

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Maibach, H. I.

M. A. Farage, K. W. Miller, P. Elsner, and H. I. Maibach, “Intrinsic and extrinsic factors in skin ageing: a review,” Int. J. Cosmet. Sci.30(2), 87–95 (2008).
[CrossRef] [PubMed]

J. M. Waller and H. I. Maibach, “Age and skin structure and function, a quantitative approach (I): blood flow, pH, thickness, and ultrasound echogenicity,” Skin Res. Technol.11(4), 221–235 (2005).
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P. J. Matts, P. J. Dykes, and R. Marks, “The distribution of melanin in skin determined in vivo,” Br. J. Dermatol.156(4), 620–628 (2007).
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S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
[CrossRef] [PubMed]

Matts, P. J.

P. J. Matts, P. J. Dykes, and R. Marks, “The distribution of melanin in skin determined in vivo,” Br. J. Dermatol.156(4), 620–628 (2007).
[CrossRef] [PubMed]

McKee, T.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Miller, K. W.

M. A. Farage, K. W. Miller, P. Elsner, and H. I. Maibach, “Intrinsic and extrinsic factors in skin ageing: a review,” Int. J. Cosmet. Sci.30(2), 87–95 (2008).
[CrossRef] [PubMed]

Milner, Y.

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Miyazawa, A.

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
[CrossRef] [PubMed]

Müller, M.

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, “3D microscopy of transparent objects using third-harmonic generation,” J. Microsc.191(3), 266–274 (1998).
[CrossRef] [PubMed]

Nakachi, K.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Nakagawa, N.

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
[CrossRef] [PubMed]

Nakamura, K.-I.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Neerken, S.

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
[CrossRef] [PubMed]

Neuman, R.

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Nuijs, T. A.

S. Neerken, G. W. Lucassen, M. A. Bisschop, E. Lenderink, and T. A. Nuijs, “Characterization of age-related effects in human skin: A comparative study that applies confocal laser scanning microscopy and optical coherence tomography,” J. Biomed. Opt.9(2), 274–281 (2004).
[CrossRef] [PubMed]

Oshimura, M.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Pellacani, G.

C. Longo, A. Casari, F. Beretti, A. M. Cesinaro, and G. Pellacani, “Skin aging: in vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy,” J. Am. Acad. Dermatol. (2011).
[CrossRef] [PubMed]

Pena, A.-M.

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Pluen, A.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Preller, A.

M. J. Koehler, S. Hahn, A. Preller, P. Elsner, M. Ziemer, A. Bauer, K. König, R. Bückle, J. W. Fluhr, and M. Kaatz, “Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network,” Exp. Dermatol.17(6), 519–523 (2008).
[CrossRef] [PubMed]

Roublevskaia, I.

A. R. Haake, I. Roublevskaia, and M. Cooklis, “Apoptosis: a role in skin aging?” J. Investig. Dermatol. Symp. Proc.3(1), 28–35 (1998).
[PubMed]

Sakai, S.

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
[CrossRef] [PubMed]

Saleh, F.

M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H. Ahmad, and J. Uitto, “Intrinsic aging vs. photoaging: a comparative histopathological, immunohistochemical, and ultrastructural study of skin,” Exp. Dermatol.11(5), 398–405 (2002).
[CrossRef] [PubMed]

Sasajima, K.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Sawabe, M.

K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
[CrossRef] [PubMed]

Schanne-Klein, M.-C.

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Seed, B.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Shieh, D. B.

Soroka, Y.

Y. Soroka, Z. Ma’or, Y. Leshem, L. Verochovsky, R. Neuman, F. M. Brégégère, and Y. Milner, “Aged keratinocyte phenotyping: morphology, biochemical markers and effects of Dead Sea minerals,” Exp. Gerontol.43(10), 947–957 (2008).
[CrossRef] [PubMed]

Springer, S.

M. J. Koehler, S. Zimmermann, S. Springer, P. Elsner, K. König, and M. Kaatz, “Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography,” Skin Res. Technol.17(4), 479–486 (2011).
[CrossRef] [PubMed]

Squier, J.

M. Müller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, “3D microscopy of transparent objects using third-harmonic generation,” J. Microsc.191(3), 266–274 (1998).
[CrossRef] [PubMed]

Sugawara, T.

S. Sakai, M. Yamanari, A. Miyazawa, M. Matsumoto, N. Nakagawa, T. Sugawara, K. Kawabata, T. Yatagai, and Y. Yasuno, “In vivo three-dimensional birefringence analysis shows collagen differences between young and old photo-aged human skin,” J. Invest. Dermatol.128(7), 1641–1647 (2008).
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Sugimoto, M.

M. Sugimoto, R. Yamashita, and M. Ueda, “Telomere length of the skin in association with chronological aging and photoaging,” J. Dermatol. Sci.43(1), 43–47 (2006).
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Sun, C.-K.

M. R. Tsai, S. Y. Chen, D. B. Shieh, P. J. Lou, and C.-K. Sun, “In vivo optical virtual biopsy of human oral mucosa with harmonic generation microscopy,” Biomed. Opt. Express2(8), 2317–2328 (2011).
[CrossRef] [PubMed]

S.-Y. Chen, S.-U. Chen, H.-Y. Wu, W.-J. Lee, Y.-H. Liao, and C.-K. Sun, “In vivo virtual biopsy of human skin by using noninvasive higher harmonic generation microscopy,” IEEE J. Sel. Top. Quantum Electron.16(3), 478–492 (2010).
[CrossRef]

S. Y. Chen, H. Y. Wu, and C.-K. Sun, “In vivo harmonic generation biopsy of human skin,” J. Biomed. Opt.14(6), 060505 (2009).
[CrossRef] [PubMed]

C. S. Hsieh, S. U. Chen, Y. W. Lee, Y. S. Yang, and C.-K. Sun, “Higher harmonic generation microscopy of in vitro cultured mammal oocytes and embryos,” Opt. Express16(15), 11574–11588 (2008).
[PubMed]

M.-C. Chan, T.-M. Liu, S.-P. Tai, and C.-K. Sun, “Compact fiber-delivered Cr:forsterite laser for nonlinear light microscopy,” J. Biomed. Opt.10(5), 054006 (2005).
[CrossRef] [PubMed]

T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, “Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser,” Scanning23(4), 249–254 (2001).
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Supatto, W.

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K.-I. Nakamura, N. Izumiyama-Shimomura, M. Sawabe, T. Arai, Y. Aoyagi, M. Fujiwara, E. Tsuchiya, Y. Kobayashi, M. Kato, M. Oshimura, K. Sasajima, K. Nakachi, and K. Takubo, “Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium,” J. Invest. Dermatol.119(5), 1014–1019 (2002).
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Figures (6)

Fig. 1
Fig. 1

(a) A schema of a 1230 nm-based HGB system adapted from a commercial confocal scanning system combined with an inverted microscope; DBS: dichroic beam splitter; CF: color filter; BF: band-pass filter; PMT: photomultiplier tube. (b) Sketch of a home-designed vacuum-pump plate used for sample stabilization; IW: imaging window; C: circular intaglio. (c) The implementation of the vacuum-pump plate which is combined with sample stage of inverted microscope.

Fig. 2
Fig. 2

A representative in vivo HGB image stack combining SHG and THG modalities from a 24-year-old Asian subject. The images were obtained from the volar forearm at different depths including the stratum corneum (a), the stratum granulosum (b), the stratum spinosum (c), and the stratum basale (d). At the stratum basale, the cytoplasm of the basal cells appeared THG-brighter (arrowheads) (d). At the dermoepidermal junction (e), collagen fibers were revealed by SHG (arrow), while the surrounding basal cells were shown by strong THG signals (arrowhead). As moving deeper inside papillary dermis (f), the borders of the basal cells (arrowheads) can also be clearly defined. THG and SHG are represented by magenta and green pseudo-colors, respectively. Scale bar = 50 μm.

Fig. 3
Fig. 3

Four representative in vivo THG images of epidermal granular cells obtained from the volar forearm of (a) 24-, (b) 47-, (c) 59-, and (d) 69-year-old volunteers. In vivo cytological analysis of (e) cellular size, (f) nuclear size, and (g) NC ratio of the granular cells showed no statistical difference between these three age groups. There was no significant correlation between (h) age and cellular size, (i) age and nuclear size, or (j) age and NC ratio. NS, no significance. Scale bar = 100 μm.

Fig. 4
Fig. 4

Four representative in vivo THG images of epidermal basal cells obtained from the volar forearm of (a) 24-, (b) 39-, (c) 56-, and (d) 74-year-old volunteers. The widened THG-dark intercellular spaces between basal keratinocytes were indicated by arrows in (b-d). Scale bar = 50 μm.

Fig. 5
Fig. 5

The cellular size (a), nuclear size (c), and NC ratio (e) of the basal cells. A significant positive correlation was shown between age and the cellular area (b; correlation coefficient r = 0.940) as well as between age and the nuclear area (d; correlation coefficient r = 0.911) of basal cells; **P < 0.0001. No significant correlation was found between age and NC ratio (f); NS, no significance.

Fig. 6
Fig. 6

The cellular (a, b) and nuclear size (c, d) of the basal cells in female and male subjects. A significant positive correlation was shown between age and the cellular area as well as between age and the nuclear area in female (a and c) and male subjects (b and d). r, correlation coefficient.

Tables (1)

Tables Icon

Table 1 Summary of the cellular area, nuclear area and N/C ratio of granular and basal cells observed by harmonic generation microscopy in three age groups

Metrics