In vivo harmonic generation biopsy of human skin.

The ability to in vivo image deep tissues noninvasively with a high resolution is strongly required for optical virtual biopsy. Higher harmonic generation microscopy, combined with second- and third-harmonic generation microscopies, is applied to 17 Asian volunteers' forearm skin. After continuous observation for 30 min, no visible damage was found. Our study proves that harmonic generation biopsy (HGB) is able to satisfy the safety requirement and to provide high penetrability (approximately 300 microm) and submicron resolution all at the same time and is a promising tool for future virtual biopsy of skin diseases. In contrast to a previous study on fixed human skin specimens, a much improved penetrability and much reduced resolution-degradation versus depth are found in this in vivo examination.

[1]  R. Wickett,et al.  Structure and function of the epidermal barrier , 2006 .

[2]  Karsten König,et al.  In vivo assessment of human skin aging by multiphoton laser scanning tomography. , 2006, Optics letters.

[3]  R. Anderson,et al.  The optics of human skin. , 1981, The Journal of investigative dermatology.

[4]  Hans C Gerritsen,et al.  Spectrally resolved multiphoton imaging of in vivo and excised mouse skin tissues. , 2007, Biophysical journal.

[5]  P. So,et al.  Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes. , 1997, Optics letters.

[6]  Chi-Kuang Sun,et al.  In vivo and ex vivo imaging of intra-tissue elastic fibers using third-harmonic-generation microscopy. , 2007, Optics express.

[7]  P. So,et al.  Confocal microscopy and multi-photon excitation microscopy of human skin in vivo. , 2001, Optics express.

[8]  Chi-Kuang Sun,et al.  Noninvasive in vitro and in vivo assessment of epidermal hyperkeratosis and dermal fibrosis in atopic dermatitis. , 2009, Journal of biomedical optics.

[9]  B R Masters,et al.  Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin. , 1997, Biophysical journal.

[10]  Chi‐Kuang Sun,et al.  Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo. , 2006, Journal of biomedical optics.

[11]  Chi-Kuang Sun,et al.  In vivo optical biopsy of hamster oral cavity with epi-third-harmonic-generation microscopy. , 2006, Optics express.

[12]  Tsung-Han Tsai,et al.  Optical biopsy of fixed human skin with backward-collected optical harmonics signals. , 2005, Optics express.

[13]  Milind Rajadhyaksha,et al.  Skin imaging with reflectance confocal microscopy. , 2008, Seminars in cutaneous medicine and surgery.

[14]  Szu-Yu Chen,et al.  Least invasive in vivo imaging using harmonic generation microscopy , 2008, SPIE BiOS.

[15]  Aernout Kisteman,et al.  Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media , 2000 .

[16]  Chi-Kuang Sun,et al.  Higher harmonic generation microscopy. , 2005, Advances in biochemical engineering/biotechnology.

[17]  Chi-Kuang Sun,et al.  Higher harmonic generation microscopy of in vitro cultured mammal oocytes and embryos. , 2008, Optics express.

[18]  Shi-Wei Chu,et al.  Wavelength dependent damage in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources , 2002 .

[19]  J. Borredon,et al.  Dynamic In Vivo Measurement of Erythrocyte Velocity and Flow in Capillaries and of Microvessel Diameter in the Rat Brain by Confocal Laser Microscopy , 1999, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[20]  Chi‐Kuang Sun,et al.  Optical signal degradation study in fixed human skin using confocal microscopy and higher-harmonic optical microscopy. , 2006, Optics express.

[21]  Masahiro Ito,et al.  Ex vivo and in vivo second-harmonic-generation imaging of dermal collagen fiber in skin: comparison of imaging characteristics between mode-locked Cr:forsterite and Ti:sapphire lasers. , 2009, Applied optics.