Non-linear fluorescence lifetime imaging of biological tissues |
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Authors: | Riccardo Cicchi Francesco Saverio Pavone |
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Institution: | (1) European Laboratory for Non-linear Spectroscopy (LENS), University of Florence, Via Nello Carrara 1, 50019 Sesto Fiorentino, Italy |
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Abstract: | In recent years fluorescence microscopy has become a widely used tool for tissue imaging and spectroscopy. Optical techniques,
based on both linear and non-linear excitation, have been broadly applied to imaging and characterization of biological tissues.
Among fluorescence techniques used in tissue imaging applications, in recent years two and three-photon excited fluorescence
have gained increased importance because of their high-resolution deep tissue imaging capability inside optically turbid samples.
The main limitation of steady-state fluorescence imaging techniques consists in providing only morphological information;
functional information is not detectable without technical improvements. A spectroscopic approach, based on lifetime measurement
of tissue fluorescence, can provide functional information about tissue conditions, including its environment, red-ox state,
and pH, and hence physiological characterization of the tissue under investigation. Measurement of the fluorescence lifetime
is a very important issue for characterizing a biological tissue. Deviation of this property from a control value can be taken
as an indicator of disorder and/or malignancy in diseased tissues. Even if much work on this topic has still to be done, including
the interpretation of fluorescence lifetime data, we believe that this methodology will gain increasing importance in the
field of biophotonics. In this paper, we review methodologies, potentials and results obtained by using fluorescence lifetime
imaging microscopy for the investigation of biological tissues. |
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