Bs 0295d cy3

Para n-embedded brain sections were rinsed in 0.01 M PBS (pH 7.3) 3 times (10 min for each) and blocked in 0.01 M PBS containing 10% normal donkey serum and 0.3% Triton X-100 for 1 h at RT. And then, the sections were incubated for 1 h at RT and for 48 h at 4°C with primary antibodies: goat anti-Iba1 antibody (1:500; ab5076, abcam, USA), mouse anti-GFAP (1:1000; MAB3402, Merck, Germany), mouse anti-NeuN (1:1000; MAB377, Merck, Germany) and rabbit anti-NEK7 antibody (1:800; NBP1-31110, NOVUS, USA) in PBS containing 0.3% (v/v) Triton X-100, 0.25% (w/v) λ-carrageenan, and 5% (v/v) donkey serum (PBS-XCD). All sections were washed three times in 0.01 M PBS (10 min each), and were then incubated for 1.5 h at RT with Donkey Anti-Goat IgG/FITC antibody (1:1000; bs-0294D-FITC, BIOSS, China), Donkey Anti-rabbit IgG/Cy3 antibody (1:1000; bs-0295D-Cy3, BIOSS, China) and Goat Anti-Mouse IgG/FITC antibody (1:1000; bs-0296G-FITC, BIOSS, China) respectively. Finally, all sections were air-dried and coverslipped with a mixture of 0.05 M PBS containing 50% (v/v) glycerin and 2.5% (w/v) triethylenediamine. The confocal images were obtained, and digital images were captured using a Fluoview laser scanning confocal microscopes (Olympus) equipped with the FV1000 (Ver.1.7a) software.