Non-linear effects and role of scattering in multiphoton imaging of thick biological samples

Francesca Cella; Zeno Lavagnino; Alberto Diaspro

doi:10.1117/12.809258

25 February 2009 Non-linear effects and role of scattering in multiphoton imaging of thick biological samples

Francesca Cella, Zeno Lavagnino, Alberto Diaspro

Proceedings Volume 7183, Multiphoton Microscopy in the Biomedical Sciences IX; 718324 (2009) https://doi.org/10.1117/12.809258
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Non linear optical scanning microscopy has became a useful tool for living tissue imaging. Biological tissues are highly scattering media and this leads to an exponential attenuation of the excitation intensity as the light travels into the sample. While performing imaging of biological scattering tissues in non linear excitation regime, the localization of the maximum 2PE intensity was found to shift closer to the surface¹ and the 2PE imaging depth limit appears strongly limited by near surface fluorescence.² In this work we computed the illumination and the photobleaching distribution³ in order to characterize the effects induced by scattering. The simulations have been performed for different scattering coefficients and different focus depth. An experimental test has been carried out by imaging, with 0.9 numerical aperture objective, thick scattering fluorescent immobile sample (polyelectrolyte gel). Results confirm that under these conditions no photobleaching effects due to scattering occur close to the surface.

Citation Download Citation

Francesca Cella, Zeno Lavagnino, and Alberto Diaspro "Non-linear effects and role of scattering in multiphoton imaging of thick biological samples", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 718324 (25 February 2009); https://doi.org/10.1117/12.809258

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