22 February 2013 Cell metabolism, tumour diagnosis and multispectral FLIM
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Fluorescence guided diagnosis of tumour tissue is in many cases insufficient, because false positive results are interfering with the outcome. Discrimination between tumour and inflammation could be therefore difficult. Improvement of fluorescence diagnosis through observation of cell metabolism could be the solution, which needs a detailed understanding of the origin of autofluorescence. However, a complex combination of fluorophores give rise to the emission signal. Also in PDD (photodynamic diagnosis) different photosensitizer metabolites contribute to the fluorescence signal. Therefore, the fluorescence decay in many cases does not show a simple monoexponential profile. In those cases a considerable improvement could be achieved when time-resolved and spectral-resolved techniques are simultaneously incorporated. The discussion will focus on the detection of NADH, FAD and 5-ALA induced porphyrins. With respect to NADH and FAD the discrimination between protein bound and free coenzyme was investigated with multispectral FLIM in normal oral keratinocytes and squamous carcinoma cells from different origin. The redox ratio, which can be correlated with the fluorescence lifetimes of NADH and FAD changed depending on the state of the cells. Most of the investigations were done in monolayer cell cultures. However, in order to get information from a more realistic in vivo situation additionally the chorioallantoismembrane (CAM) of fertilized eggs was used where tumour cells or biopsies were allowed to grow. The results of theses measurements will be discussed as well.
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A. Rück, A. Rück, C. Hauser, C. Hauser, S. Lorenz, S. Lorenz, S. Mosch, S. Mosch, S. Rotte, S. Rotte, M. Kessler, M. Kessler, S. Kalinina, S. Kalinina, "Cell metabolism, tumour diagnosis and multispectral FLIM ", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85880U (22 February 2013); doi: 10.1117/12.2003729; https://doi.org/10.1117/12.2003729

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