Two-photon fluorescence and second-harmonic generation imaging of collagen in human tissue based on multiphoton microscopy.

Multiphoton microscopic imaging of collagen plays an important role in noninvasive diagnoses of human tissue. In this study, two-photon fluorescence and second-harmonic generation (SHG) imaging of collagen in human skin dermis and submucosa of colon and stomach tissues were investigated based on multiphoton microscopy (MPM). Our results show that multiphoton microscopic image of collagen bundles exhibits apparently different pattern in human tissues. The collagen bundles can simultaneously reveal its SHG and two-photon excited fluorescence images in the submucosa of colon and stomach, whereas it solely emit SHG signal in skin dermis. The intensity spectral information from tissues further demonstrated the above results. This indicates that collagen bundles have completely different space arrangement in these tissues. Our experimental results bring more detailed information of collagen for the application of MPM in human noninvasive imaging.

[1]  H. S. de Bruijn,et al.  In vivo nonlinear spectral imaging in mouse skin. , 2006, Optics express.

[2]  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.

[3]  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.

[4]  Vladimir Hovhannisyan,et al.  Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy. , 2010, Journal of biomedical optics.

[5]  Josef Bille,et al.  Second harmonic generation imaging of collagen fibrils in cornea and sclera. , 2005, Optics express.

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

[7]  Shuangmu Zhuo,et al.  Spectral characteristics of autofluorescence and second harmonic generation from ex vivo human skin induced by femtosecond laser and visible lasers. , 2007, Scanning.

[8]  Karsten König,et al.  Clinical multiphoton tomography , 2008, Journal of biophotonics.

[9]  Chung-Ho Sun,et al.  Nondestructive imaging of live human keloid and facial tissue using multiphoton microscopy. , 2008, Archives of facial plastic surgery.

[10]  S. Xie,et al.  Nonlinear spectral imaging of human hypertrophic scar based on two‐photon excited fluorescence and second‐harmonic generation , 2009, The British journal of dermatology.

[11]  Anthony M. D. Lee,et al.  Spectroscopic characterization and microscopic imaging of extracted and in situ cutaneous collagen and elastic tissue components under two‐photon excitation , 2009, Skin research and technology : official journal of International Society for Bioengineering and the Skin (ISBS) [and] International Society for Digital Imaging of Skin (ISDIS) [and] International Society for Skin Imaging.

[12]  Bruce J Tromberg,et al.  Nonlinear optical microscopy of articular cartilage. , 2005, Osteoarthritis and cartilage.

[13]  K. Badizadegan,et al.  NAD(P)H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes. , 2002, Cancer research.