Journal: Nature immunology
Article Title: Fibrin-targeting immunotherapy protects against neuroinflammation and neurodegeneration
Figure Lengend Snippet: In vivo imaging of fibrinogen leakage in 5XFAD mice. a , Fluorescent Aβ probe methoxy-X04-positive amyloid plaque in a cortical section from a 3-month-old 5XFAD mouse co-labeled for fibrinogen (red) and CD11b (green). Scale bar: 20 μm. n = 4 mice. Representative images are shown. b , In vivo two-photon imaging of cortex from Thy1- YFP:5XFAD mice at 11 months of age shows Alexa594-conjugated fibrinogen (administered intravenously) (red), a methoxy-X04-positive Aβ plaque (blue), and dystrophic neurites (green, swollen green structures). Arrowheads indicate fibrinogen (red) surrounding a Aβ plaque (blue) at areas of neuritic loss. Arrows indicate fibrinogen extravasation at areas of dystrophic neurites (yellow) in proximity to a plaque (blue). Scale bars: 10 μm. (n = 4 Thy1- YFP : 5XFAD mice and n = 2 Thy1- YFP mice. Representative images are shown. c , 5B8 in vivo target engagement in the brain of 5XFAD mice. Brain sections of 5XFAD i.p.-injected with biotinylated 5B8 were stained with Cy3-streptavidin (red), anti-fibrinogen (green), and methoxy-X04-positive Aβ plaque (blue). Orthogonal views of the y/z and x/z planes show localization of fibrinogen and 5B8 antibody around amyloid plaques. Scale bar, 10 μm. n =3 mice Representative images are shown.
Article Snippet: Sections were fixed with 4% PFA for 10 min and biotinylated 5B8 was detected using Cy3-conjugated streptavidin (1:100; Invitrogen) for 30 min at 25 ˚C.
Techniques: In Vivo Imaging, Mouse Assay, Labeling, In Vivo, Imaging, Injection, Staining