Abstract

Abstract: In this work, we demonstrate the application of multiphoton polarization imaging in resolving the structures in surface stratum corneum and dermal layers of ex-vivo human skin. By varying the excitation and emission polarizations, we characterized the structural features in both Laurdan labeled stratum corneum and dermal fibers. The results presented here have important consequences in bioimaging applications of the skin. Both the mechanics of transdermal drug delivery across the skin and physiological significance of the structural changes of the dermis can be monitored. Our results show that the transition dipoles of Laurdan molecules are preferentially oriented normal to the membrane surface. Furthermore, polarization imaging shows that fibrous structures in the dermis generate emission aligned strongly along the excitation polarization. This work shows that multiphoton polarization imaging can be a powerful method in identifying structural orientations in the skin and other biological structures.

© 2003 Optical Society of America

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References

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    [CrossRef]
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Annual Review of Biomedical Engineering (1)

P. T. C. So, C. Y. Dong, B. R. Masters, and K. M. Berland, �??Two-photon excitation fluorescence microscopy,�?? Annual Review of Biomedical Engineering 2, 399-429 (2000).
[CrossRef]

Biophys. J. (1)

T. Parasassi, G. De Stasio, G. Ravagnan, R. M. Rusch, and E. Gratton, �??Quantitation of lipid phases in phospholipids vescicles by the generalized polarization of Laurdan fluorescence,�?? Biophys. J. 60, 179-189 (1991).
[CrossRef] [PubMed]

Biophysical J. (2)

L. A. Bagatolli and E. Gratton, �??Two photon fluorescence microscopy of coexisting lipid domains in giant unilamellar vesicles of binary phospholipids mixtures,�?? Biophysical J. 78, 290-305 (2000).
[CrossRef]

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, �??Polarization-modulated second harmonic generation in collagen,�?? Biophysical J. 82, 3330-3342 (2002).
[CrossRef]

J. Biomed. Opt. (1)

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L. B. Da Silva, �??Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,�?? J. Biomed. Opt. 7, 205-214 (2002).
[CrossRef] [PubMed]

J. Investigative Dermatology (3)

B. Yu, C. Y. Dong, P. T. C. So, D. Blankschtein, and R. Langer, �??In vitro visualization and quantification of oleic acid induced changes in transdermal transport using two-photon fluorescence microscopy,�?? J. Investigative Dermatology 117, 16-25 (2001).
[CrossRef]

B. Yu, K. H. Kim, P. T. C. So, D. Blankschtein, and R. Langer, �??Topographic heterogeneity in transdermal transport revealed by high-speed two-photon microscopy: determination of representative skin sample sizes,�?? J. Investigative Dermatology 118, 1085-1088 (2002).
[CrossRef]

B. Yu, K. H. Kim, P. T. C. So, D. Blankschtein, and R. Langer, �??Visualization of oleic-acid induced transdermal diffusion pathways using two-photon fluorescence microscopy,�?? J. Investigative Dermatology 120, 448-455 (2003).
[CrossRef]

Opt. Lett. (1)

Science (1)

W. Denk, J. H. Strickler, and W. W. Webb, �??Two-photon laser scanning fluorescence microscopy,�?? Science 248, 73-76 (1990).
[CrossRef] [PubMed]

Other (1)

J. R. Lakowicz, Principles of fluorescence spectroscopy (Kluwer Academic/Plenum Publishers, 1999) Chaps. 10,11,12.

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Figures (4)

Fig. 1
Fig. 1

A multiphoton polarization microscope

Fig. 2.
Fig. 2.

Polarization-resolved images of laurdan treated stratum corneum (Imax=150, Imin=0)

Fig. 3.
Fig. 3.

Multiphoton polarization imaging of skin dermal fiber layers (Imax=100, Imin=0)

Table. 1.
Table. 1.

Polarization values of Laurdan labeled stratum corneum and dermal fibers

Equations (1)

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p = I II I I II + I and r = I II I I II + 2 I ,

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