In vivo nonlinear spectral imaging in mouse skin.

We report on two-photon autofluorescence and second harmonic spectral imaging of live mouse tissues. The use of a high sensitivity detector and ultraviolet optics allowed us to record razor-sharp deep-tissue spectral images of weak autofluorescence and short-wavelength second harmonic generation by mouse skin. Real-color image representation combined with depth-resolved spectral analysis enabled us to identify tissue structures. The results show that linking nonlinear deep-tissue imaging microscopy with autofluorescence spectroscopy has the potential to provide important information for the diagnosis of skin tissues.

[1]  B R Masters,et al.  Multiphoton Excitation Microscopy of In Vivo Human Skin: Functional and Morphological Optical Biopsy Based on Three‐Dimensional Imaging, Lifetime Measurements and Fluorescence Spectroscopy a , 1998, Annals of the New York Academy of Sciences.

[2]  Jianan Y Qu,et al.  Two-photon autofluorescence spectroscopy and second-harmonic generation of epithelial tissue. , 2005, Optics letters.

[3]  Iris Riemann,et al.  Compact multiphoton/single photon laser scanning microscope for spectral imaging and fluorescence lifetime imaging. , 2006, Scanning.

[4]  B Miedema,et al.  Emission spectra of colonic tissue and endogenous fluorophores. , 1998, The American journal of the medical sciences.

[5]  B. Tromberg,et al.  Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Roodenburg,et al.  Autofluorescence and diffuse reflectance spectroscopy for oral oncology , 2005, Lasers in surgery and medicine.

[7]  Rebecca Richards-Kortum,et al.  Realistic three-dimensional epithelial tissue phantoms for biomedical optics. , 2002, Journal of biomedical optics.

[8]  A. Pena,et al.  Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy. , 2005, Optics express.

[9]  William A Mohler,et al.  Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. , 2002, Biophysical journal.

[10]  J. Kennedy,et al.  The nature of the chromophore responsible for naturally occurring fluorescence in mouse skin. , 1988, Journal of photochemistry and photobiology. B, Biology.

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

[12]  Peng Xi,et al.  Depth-resolved fluorescence spectroscopy of normal and dysplastic cervical tissue. , 2005, Optics express.

[13]  R. Pepperkok,et al.  Spectral imaging and linear un‐mixing enables improved FRET efficiency with a novel GFP2–YFP FRET pair , 2002, FEBS letters.

[14]  Melissa C Skala,et al.  Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues. , 2005, Cancer research.

[15]  W. Webb,et al.  Three‐dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two‐photon excitation laser scanning microscopy , 1995, Journal of microscopy.

[16]  R R Alfano,et al.  Subsurface tumor progression investigated by noninvasive optical second harmonic tomography. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  E. Epstein,et al.  Human skin collagen. Presence of type I and type III at all levels of the dermis. , 1978, The Journal of biological chemistry.

[18]  P. So,et al.  Two-photon 3-D mapping of ex vivo human skin endogenous fluorescence species based on fluorescence emission spectra. , 2005, Journal of biomedical optics.

[19]  Massoud Motamedi,et al.  In vivo multimodal nonlinear optical imaging of mucosal tissue. , 2004, Optics express.

[20]  Frédérique Frouin,et al.  FRET multiphoton spectral imaging microscopy of 7-ketocholesterol and Nile Red in U937 monocytic cells loaded with 7-ketocholesterol. , 2004, Analytical and quantitative cytology and histology.

[21]  A Mateasik,et al.  Spectral unmixing of flavin autofluorescence components in cardiac myocytes. , 2005, Biophysical journal.

[22]  Yasushi Hiraoka,et al.  Spectral imaging fluorescence microscopy , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[23]  Steven L. Jacques,et al.  In vivo fluorescence spectroscopy and imaging of human skin tumors. , 1995 .

[24]  W. Webb,et al.  Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Schmid,et al.  Application of spectral imaging microscopy in cytomics and fluorescence resonance energy transfer (FRET) analysis , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[26]  T J Flotte,et al.  Ultraviolet laser‐induced fluorescence of colonic tissue: Basic biology and diagnostic potential , 1992, Lasers in surgery and medicine.

[27]  David W Piston,et al.  Quantitative NAD(P)H/Flavoprotein Autofluorescence Imaging Reveals Metabolic Mechanisms of Pancreatic Islet Pyruvate Response* , 2004, Journal of Biological Chemistry.

[28]  S. L. Jacques,et al.  In vivo fluorescence spectroscopy and imaging of human skin tumours , 1994, Lasers in Medical Science.

[29]  Brian Seed,et al.  Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation , 2003, Nature Medicine.

[30]  R. Richards-Kortum,et al.  Study of the fluorescence properties of normal and neoplastic human cervical tissue , 1993, Lasers in surgery and medicine.