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Immunofluorescence and Thioflavin S (ThioS) staining

QS Qiang Su
TL Tian Li
PH Pei-Feng He
XL Xue-Chun Lu
QY Qi Yu
QG Qi-Chao Gao
ZW Zhao-Jun Wang
MW Mei-Na Wu
DY Dan Yang
JQ Jin-Shun Qi
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Brain slices were washed in PBS, permeabilized for 15 min by shaking at room temperature with PBS containing 0.5% Triton X-100, rinsed in PBS, and then blocked with 5% bovine serum albumin (BSA; AR1006, BOSTER) in PBS for 1 h at room temperature. Thereafter, sections were incubated overnight at 4 °C with primary antibody and then placed in a wet box containing a little water. After rinsing, sections were incubated with appropriate fluorescence-conjugated secondary antibodies at room temperature for 1 h. For ThioS staining, sections were stained with 0.05% ThioS (23059, AAT Bioquest) in 50% ethanol in dark for 8 min at room temperature, followed by two rinsing in 80% ethanol for 10 s each. Finally, DAPI (C0065, Solarbio) was used for nuclear staining. Slides were then sealed with the fluorescent mounting medium (AR1109, BOSTER). Immunofluorescence images were acquired using a fluorescent microscope (Olympus) and analyzed using the ImageJ software.

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