The slices was performed at 3 sites, each 400 ?340 m. The brain sections > 커뮤니티 카카오소프트 홈페이지 방문을 환영합니다.

The slices was performed at 3 sites, each 400 ?340 m. The brain sections were then individually picked up using a paint brush and were transferred to a well filled with 0.1M PBS. Slices were then either stained immediately following the appropriate protocol or stored in a cryoprotectant solution at -20 until further use.Staining methods Lectin staining3.5 and 6 month old rTg4510 mice were injected intracranially with the fibrin 377-395 peptide-conjugated -Fe 377-395 peptide (1:3 2O3 nanoparticles and free fibrin weight ratio). Briefly, mice were anaesthetized before PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/13485127 surgery with a ketamine:xylazine at 150 and 12 mg/kg, respectively [53]. The animals were then placed into a stereotaxic apparatus with a heating pad to maintain body temperature. Stereotaxic coordinates were bregma + 1.5 mm anterior-posterior, 1.5 mm lateral and 1.8 mm vertical for frontal cortex. Subsequently, 10 L of the fibrin 377-395 peptide-conjugated -Fe2O3 nanoparticles dispersed in PBS (2 mg/ml, 10 ug bound fibrin 377-395 per injection) and free fibrin 377-395 peptide dissolved in a PBS solution (3 mg/ml), were intracranially injected in the right and left hemispheres, respectively. The 4-Bromo-5-nitro-1H-indazole injection holes were then filled with bone wax. The mice were stitched and then returned to their cages. All mice used in our experiments were then sacrificed 30 daysThe slices were first introduced into a PBS solution in order to remove the OCT from the tissue. The slices were treated with FITC-conjugated Bandeiraea simplicifolia isolectin B4 (IB-4; 1:50; Sigma-Aldrich) for 1 h, and then were washed three times for 5 min each in PBS. The slices were then mounted on slides and stored in the dark to air-dry. A drop of DAPI (1:2000; SigmaAldrich) solution was added to sections after drying using a Pasteur pipette for 1 min and washed twice for 1 min each in PBS. The slides were then sealed using a cover slip.Iron stainingThe slices were mounted on slides and placed in 2 potassium ferrocyanide solution with 2 HCl solution for 30 min. Afterwards, the slides were washed three times in distilled water and sealed with a cover slip.Glat et al. Journal of Nanobiotechnology 2013, 11:32 http://www.jnanobiotechnology.com/content/11/1/Page 11 ofImmunohistochemistryThe slices were first introduced into a PBS solution in order to remove the OCT from the tissue. The slices were incubated in 0.5 Triton for 20 min and then washed three times for 5 min each in PBS. The slices were then incubated in a blocking solution (NGS, 5 in PBS; from Jackson Immuno Research) for 1 h. After the blocking stage, slices were treated with Anti-HumanPHF-Tau Monoclonal Antibody Ethyl (4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)carbamate hydrochloride (AT8; 1:1000 in normal goat serum; Thermo Scientific) and then left overnight at 4 . Following incubation, the slices were washed three times for 5 min each in PBS. The slices were treated with Cy3 donkey anti-mouse (Cy3 1:500, Jackson Immuno Research). PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/3971254 After 1 h, the slices were washed three times in PBS. The slices were then mounted on slides, stored in the dark to air-dry and then sealed using a cover slip. If NFTs staining was performed as well, a drop of 0.05 Thio-S solution (Sigma-Aldrich) was added to sections after drying using a Pasteur pipette. The sections were then incubated for 8 min in the dark, followed by two washes for 10 sec in 80 ethanol solution and one wash in distilled water. The slides were then stored in the dark to air-dry and sealed using a cover slip. To study microglia morphology, Iba-1 staining was performed. T.