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

R. Cicchi; D. Massi; D. Stambouli; D. D. Sampson; F. S. Pavone

doi:10.1117/12.644193

23 February 2006 Contrast enhancement in combined two-photon second harmonic imaging of skin by using hyperosmotic agents

R. Cicchi, D. Massi, D. Stambouli, D. D. Sampson, F. S. Pavone

Author Affiliations +

Proceedings Volume 6089, Multiphoton Microscopy in the Biomedical Sciences VI; 60890X (2006) https://doi.org/10.1117/12.644193
Event: SPIE BiOS, 2006, San Jose, California, United States

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.

Citation Download Citation

R. Cicchi, D. Massi, D. Stambouli, D. D. Sampson, and F. S. Pavone "Contrast enhancement in combined two-photon second harmonic imaging of skin by using hyperosmotic agents", Proc. SPIE 6089, Multiphoton Microscopy in the Biomedical Sciences VI, 60890X (23 February 2006); https://doi.org/10.1117/12.644193

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available