For Immunoelectron microscopy experiment, purified exosomes were pipetted onto carbon-coated TEM grids and incubated for 5 min at room temperature. The solution was then wicked off the grid with Whatman 1 filter paper until a thin film remained. Rabbit anti-Alix antibody (exosome marker) and mouse anti-Tau12 antibody were used to label exosomes and tau, respectively. Goat anti-rabbit secondary antibody conjugated to 10 nm gold particles (Aurion, Wageningen, the Netherlands) and goat anti-mouse secondary antibody conjugated to 5nm gold particles were used for detection. The grids were then negatively stained by placement on droplets of 2% uranyl acetate for 2 min. Samples were imaged in a Hitachi H7500 transmission electron microscope equipped with an Advanced Microscopy Sciences XR60 camera.
For PHF imaging, PHF derived from rTg4510 mice brain were pipetted onto carbon-coated TEM grids and incubated for 5 min at room temperature. The solution was then wicked off the grid with Whatman 1 filter paper until a thin film remained. The grids were then negatively stained by placement on droplets of 2% uranyl acetate for 2 min. Samples were imaged in a Hitachi H7500 transmission electron microscope equipped with an Advanced Microscopy Sciences XR60 camera.
For PHF imaging, PHF derived from rTg4510 mice brain were pipetted onto carbon-coated TEM grids and incubated for 5 min at room temperature. The solution was then wicked off the grid with Whatman 1 filter paper until a thin film remained. The grids were then negatively stained by placement on droplets of 2% uranyl acetate for 2 min. Samples were imaged in a Hitachi H7500 transmission electron microscope equipped with an Advanced Microscopy Sciences XR60 camera.