23 February 2018 In vivo multiphoton and fluorescence lifetime imaging microscopy of the healthy and cholestatic liver
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A cholestatic liver disease presents one of the most common liver diseases and can potentially progress to cirrhosis or even cholangiocarcinoma. Conventional techniques are insufficient to precisely describe the complex internal structure, heterogeneous cell populations and the dynamics of biological processes of the liver. Currently, the methods of multiphoton and fluorescence lifetime imaging microscopy are actively introducing to biomedical research. Those methods are extremely informative and non-destructive that allows studying of a large number of processes occurring inside cells and tissues, analyzing molecular cellular composition, as well as evaluating the state of connective tissue fibers due to their ability to generate a second optical harmonic. Multiphoton and FLIM microscopy do not need additional staining of samples or the incorporation of any markers to study metabolism, lipid composition, microstructure analysis, evaluation of fibrous structures. These parameters have pronounced changes in hepatocytes of liver with common pathological diseases. Thereby in this study we investigated metabolic changes in the healthy and cholestatic liver based on the fluorescence of the metabolic co-factors NAD(P)H and FAD by multiphoton microscopy combined with FLIM. To estimate the contribution of energy metabolism and lipogenesis in the observed changes of the metabolic profile, a separate analysis of NADH and NADPH was presented. The data can be used to develop new criteria for the identification of hepatic pathology at the level of hepatocyte changes directed to personalized medicine in the future.
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Daria S. Kuznetsova, Daria S. Kuznetsova, Varvara V. Dudenkova, Varvara V. Dudenkova, Svetlana A. Rodimova, Svetlana A. Rodimova, Nikolai V. Bobrov, Nikolai V. Bobrov, Vladimir E. Zagainov, Vladimir E. Zagainov, Elena V. Zagaynova, Elena V. Zagaynova, } "In vivo multiphoton and fluorescence lifetime imaging microscopy of the healthy and cholestatic liver", Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 1049827 (23 February 2018); doi: 10.1117/12.2292315; https://doi.org/10.1117/12.2292315

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