Cy3 conjugated goat anti mouse igg h l

The rats were anesthetized and perfused with 4% paraformaldehyde, then the brain tissues were collected, fixed and sliced. Next, 10% normal donkey or goat blocking serum (Solarbio, Beijing, China) was used to block brain tissues sections for 30 min at 37°C. The sections (10 μm) were then incubated at 4°C overnight with the following primary antibodies: TSPO (1:100 dilution, pa5-19088; ThermoFish Scientific), Iba1 (1:200 dilution, gb12105; Servicebio), ATG7 (1:100 dilution, ab133528; Abcam), LC3B (1:100 dilution, ab192890; Abcam) and p62 (1:100 dilution, ab109012; Abcam). The secondary antibodies Cy3 conjugated Donkey Anti-Mouse IgG (H+L) (1:200 dilution, gb21401; Servicebio), FITC conjugated Donkey Anti-Goat IgG (H+L) (1:200 dilution, gb22404; Servicebio), Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG (H+L) (1:400 dilution, gb25303; Servicebio) and Cy3 conjugated Goat Anti-mouse IgG (H+L) (1:300 dilution, gb21301; Servicebio) were used and incubated 1 h in the dark. The sections were incubated with Hoechst (1:600 dilution, g1011; Servicebio) for 3 min for nuclear staining. The sections were analyzed and photographed using a laser scanning confocal microscope (NIKON Eclipse Ti, Japan). Finally, Image J performs quantitative analysis.

Sections were routinely dewaxed and dehydrated. The sections were permeated with 0.5% Triton X-100 after repair of antigen, then incubated with 10% goat serum for 1 h. The samples were incubated with primary antibodies, rat anti-iNOS (1:200, Servicebio, China), and mice anti-Iba-1 (1:200, Servicebio, China) at 4 °C overnight. The samples were also incubated with Alexa Fluor 488 conjugated Goat Anti-rabbit IgG (H⁺L) (1:500, Servicebio, China) and Cy3 conjugated Goat Anti-mouse IgG (H⁺L) (1:300, Servicebio, China) secondary antibodies for 1 h. The sections were also incubated with DAPI for 5 min and visualized via fluorescence microscopy (Leica DM IL LED microscope, Wetzlar, Germany). Eight randomly selected fields were used to calculate the average fluorescence intensity of iNOS and Iba-1 via ImageJ 1.5.1 software.

The PZT was obtained from SCH Technology Co. Ltd. The 443 nm μ-LEDs were obtained from Shenzhen Rico Photoelectric Technology Co., Ltd. The red μ-LEDs were obtained from Shenzhen Super high-brightness Electronics Co., Ltd. The following reagents and antibodies were used in the study: Anti- IBA 1 Mouse mAb (Servicebio, GB12105; diluted: 1:100), Cy3 conjugated Goat Anti-mouse IgG (H + L) (Servicebio, GB21301; diluted: 1:100), Anti- GFAP Rabbit pAb (Servicebio, GB11096; diluted: 1:100), fluorescein isothiocyanate (FITC)–conjugated goat anti-rabbit IgG secondary antibody (Servicebio, GB22303; diluted: 1:100), TMR (Red) Tunel Cell Apoptosis Detection Kit (Servicebio, G1502-50T; Recombinant TdT enzyme: TMR-5-dUTP Labeling Mix: Equilibration Buffer (1: 5: 50)), Hematoxylin (Servicebio, G1004), Eosin Y (Biobomei, YE2080), 4′,6-diamidino-2-phenylindole (Servicebio, G1012), Toluidine blue (Servicebio, G1032).