Physical and biological limitations of in vivo multiphoton fluorescence microscopy

Alexey I. Korytin; Ekaterina A. Sergeeva; Tatiana Shcherbatyuk; Irina Parfenova

doi:10.1117/12.593410

30 March 2005 Physical and biological limitations of in vivo multiphoton fluorescence microscopy

Alexey I. Korytin, Ekaterina A. Sergeeva, Tatiana Shcherbatyuk, Irina Parfenova

Proceedings Volume 5700, Multiphoton Microscopy in the Biomedical Sciences V; (2005) https://doi.org/10.1117/12.593410
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

In this paper we present experimental and theoretical study of limitations on deep imaging of biological tissue structure using multiphoton fluorescence microscopy (MFM), that are imposed by scattering of short NIR pulses in optically turbid medium. The time stretching of the laser pulse and its transversal widening limit the maximum imaging depth of MFM since the magnitude of fluorescence response from the medium is the power function of energy flux density in the excitation laser beam. Increasing the imaging depth by raising the laser power may result in phototoxic damage of biological objects. We have theoretically and experimentally studied temporal structure of collimated femtosecond laser pulse scattered in an optically turbid model medium with controlled concentration of micron-sized spherical beads. In parallel we investigated phototoxic effect on biological liquid at high-intense femtosecond NIR pulse irradiation for various expositions by structural analysis of dehydrated blood plasma droplet. The irradiation regimes were found with complete recovery of the droplet structure, as well as with its slight and severe stable modification.

Citation Download Citation

Alexey I. Korytin, Ekaterina A. Sergeeva, Tatiana Shcherbatyuk, and Irina Parfenova "Physical and biological limitations of in vivo multiphoton fluorescence microscopy", Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, (30 March 2005); https://doi.org/10.1117/12.593410

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KEYWORDS

Scattering

Femtosecond phenomena

Laser scattering

Light scattering

Plasma

Near infrared

Blood

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