8-10 µm frozen sections were washed in PBT (PBS + 1% Triton-X), blocked in 5% Normal Donkey Serum in PBT, and incubated overnight at 4 °C with primary antibodies and detected with species-specific secondary antibodies coupled to Alexa Fluor 488, 555, 594, and 647 (Life Technologies) for 1 h at room temperature.
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