We stained coronal free-floating sections with primary antibodies against pser129 (pser129, rabbit, 1:5000; Abcam, Ab51253) and NeuN (mouse, 1:1000, Millipore, Mab377), and secondary antibodies AlexaFluor 488 anti-mouse (goat, 1:500, Jackson Immunoresearch Laboratories, 115-545-146) and AlexaFluor 594 goat anti-rabbit (goat, 1:500, Jackson Immunoresearch Laboratories, 111-585-144). The staining with thioflavin S was performed in accordance with our previous work [59 (link)].
Stained slides were blind coded before analysis and were imaged with an inverted confocal laser microscope Nikon Eclipse Ti-E. Thioflavin S staining was imaged by averaging 4x scans for each z step of the stack to remove noise using NIS Elements AR 4.00.08 software (Nikon). For all other fluorescence staining, we acquired images by single scan per stack-step. We post-treated the confocal stacks with a kernel-3 median filter to remove noise and generated all final images using NIS Elements AR 4.00.08 software (Nikon).
Stained slides were blind coded before analysis and were imaged with an inverted confocal laser microscope Nikon Eclipse Ti-E. Thioflavin S staining was imaged by averaging 4x scans for each z step of the stack to remove noise using NIS Elements AR 4.00.08 software (Nikon). For all other fluorescence staining, we acquired images by single scan per stack-step. We post-treated the confocal stacks with a kernel-3 median filter to remove noise and generated all final images using NIS Elements AR 4.00.08 software (Nikon).
