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Contrast enhancement in combined two photon - Second harmonic imaging of skin by using hyperosmotic agents

Cicchi, R., Massi, D., Stambouli, D., Sampson, David and Pavone, F.S. (2006) Contrast enhancement in combined two photon - Second harmonic imaging of skin by using hyperosmotic agents

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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.

Item Type: Conference or Workshop Item (UNSPECIFIED)
Divisions : Faculty of Engineering and Physical Sciences
Faculty of Health and Medical Sciences
Authors :
Cicchi, R.
Massi, D.
Stambouli, D.
Pavone, F.S.
Date : 2006
DOI : 10.1117/12.644193
Uncontrolled Keywords : Light propagation in tissue, Multiphoton microscopy, Second harmonic generation, Tissue imaging, Fluorescence, Image enhancement, Light propagation, Medical imaging, Second harmonic generation, Skin, Tissue, Light propagation in tissue, Multiphoton microscopy, Tissue imaging, Contrast media
Depositing User : Maria Rodriguez-Marquez
Date Deposited : 11 Jun 2018 08:46
Last Modified : 19 Sep 2018 11:32

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