Characterization of the metabolic conditions of different tissues during Zebrafish development by non-linear microscopy
Authors: Giubani C., Mercatelli R., Vanzi F., Cicchi R., Pavone F.S.
Autors Affiliation: European Laboratory for Non-Linear Spectroscopy, Sesto Fiorentino, Italy; National Institute of Optics, National Research Council (INO-CNR), Sesto Fiorentino, Italy; Department of Biology, Sesto Fiorentino, Italy; Department of Physics, University of Florence, Sesto Fiorentino, Italy
Abstract: Zebrafish is a model organism in developmental biology for several reasons, including the fact that Zebrafish embryos are translucent and this enables the study of tissue development under a microscope. This study is aimed at developing methodologies for characterizing the metabolic state of different tissues during development and in different controlled environmental conditions. Two-photon excited fluorescence (TPEF) microscopy and Fluorescence Lifetime Imaging Microscopy (FLIM) were used to study different tissues during embryos development and to characterize them in various controlled environmental conditions. The extensive anatomical mapping of the Zebrafish embryos was obtained through the detection of autofluorescence. Differences in metabolism among different organs were observed by measuring NADH/FAD ratio and free NADH/bound NADH ratio. The proposed method has the potential for monitoring embryo metabolism in different conditions during development.
KeyWords: Biology; Fluorescence; Fluorescence microscopy; Histology; Metabolism; Physiology; Tissue, Developmental biology; Environmental conditions; FLIM; Fluorescence lifetime imaging microscopy; NADH; TPEF; Two-photon excited fluorescence microscopies; Zebrafish, Tissue engineeringDOI: 10000000000