Contrast enhancement in combined two photon – second harmonic imaging of skin by using hyperosmotic agents

Year: 2006

Authors: Cicchi R., Massi D., Stambouli D., Sampson D.D., Pavone FS.

Autors Affiliation: L.E.N.S. & Department of Physics, University of Florence, G. Sansone 1, 50019 Sesto Fiorentino, Florence, Italy;
Department of Human Pathology and Oncology, University of Florence, G.B.Morgagni 85, 50134 Florence, Italy;
L.E.N.S., University of Florence, N. Carrara 1, 50019 Sesto Fiorentino, Florence, Italy;
Optical+Biomedical Engineering Laboratory, School of Electrical, Electronic & ComputerEngineering The University of Western Australia, Australia

Abstract: We used combined simultaneous two-photon excitation fluorescence microscopy (TPE) and second harmonic generation microscopy (SHG) on human skin tissue slices. We studied the effect caused by topical application of optical clearing agents (OCAs). We demonstrated that hyperosmotic agents as glycerol, propylene glycol and glucose in aqueous solution, are all effective in improving excitation light penetration depth and in enhancing image contrast. The effect caused on acquired images by sample immersion in OCAs or in their aqueous dilution, was studied. We observed a similar clearing effect with TPE and SHG acquisitions, with different effectiveness and rising time for each agent. The TPE acquired data are in good agreement with a simple diffusion model developed. From the SHG acquisition some different behaviour was observed. All three agents are potentially bio-compatible and effective in reducing scattering, improving light penetration depth and image contrast. Use of OCA can be suitable for in vivo application in two-photon microscopy, as well as in other techniques performing optical biopsy of human skin tissue.

Conference title: Saratov Fall Meeting 2005
Place: Saratov, Russia

More Information: We thank LENS (European Laboratory for Non-linear Spectroscopy), Contract Number RII3-CT2003- 506350 and Ente Cassa di Risparmio di Firenze for funding this project. David Sampson thanks LENS and Department of Physics, University of Florence for his sabbatical stay.
KeyWords: Fluorescence; Image enhancement; Light propagation; Medical imaging; Second harmonic generation; Skin; Tissue, Light propagation in tissue; Multiphoton microscopy; Tissue imaging, Contrast media
DOI: 10.1117/12.644193

Citations: 1
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