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Title: 2D polarization imaging as a low-cost fluorescence method to detect α-synuclein aggregation ex vivo in models of Parkinson’s disease
Authors: Camacho, RafaelTäuber, DanielaHansen, ChristianShi, JuanziBousset, LucMelki, RonaldLi, Jia-YiScheblykin, Ivan G.
Publishers Version: https://doi.org/10.1038/s42003-018-0156-x
Issue Date: 2018
Published in: Communications Biology 1 (2018)
Publisher: Berlin : Nature Publishing
Abstract: A hallmark of Parkinson’s disease is the formation of large protein-rich aggregates in neurons, where α-synuclein is the most abundant protein. A standard approach to visualize aggregation is to fluorescently label the proteins of interest. Then, highly fluorescent regions are assumed to contain aggregated proteins. However, fluorescence brightness alone cannot discriminate micrometer-sized regions with high expression of non-aggregated proteins from regions where the proteins are aggregated on the molecular scale. Here, we demonstrate that 2-dimensional polarization imaging can discriminate between preformed non-aggregated and aggregated forms of α-synuclein, and detect increased aggregation in brain tissues of transgenic mice. This imaging method assesses homo-FRET between labels by measuring fluorescence polarization in excitation and emission simultaneously, which translates into higher contrast than fluorescence anisotropy imaging. Exploring earlier aggregation states of α-synuclein using such technically simple imaging method could lead to crucial improvements in our understanding of α-synuclein-mediated pathology in Parkinson’s Disease.
Keywords: Parkinson’s disease; protein-rich aggregates; α-synuclein
DDC: 570
License: CC BY 4.0 Unported
Link to License: https://creativecommons.org/licenses/by/4.0/
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