Methylbenzene

Terminal transferase-mediated biotin 2′-deoxyuridine, 5′-triphosphate nick end labeling (TUNEL) assay was performed to evaluate apoptosis of cells with a TUNEL assay kit (ab206386, Abcam, Cambridge, UK) as previously described (27 (link)). The hippocampus tissue was first deparaffinized and treated with 20 μg/ml deoxyribonuclease-free proteinase K (AM2548, Thermo Fisher Scientific, Waltham, MA, USA) at room temperature for 20 min and was then washed with PBS three times (5 min for each). After being fixed at room temperature for 15 min, the samples were washed with PBS for another three times (5 min each time). Next, each sample was added with 50 μl of biotin labeling and incubated for 1 h at 37°C, followed by processing with a 3,3′-diaminobenzidine kit (DA1015, Solarbio, Beijing, China) and PBS washing. Then, the samples were sealed after clearing with methylbenzene (244511, Sigma-Aldrich, USA) for 5 min. The apoptotic cells were observed using a stereomicroscope (SZX10, Olympus, Japan) under × 100 and × 200 magnifications.

Silver nitrate (99%), sodium citrate (99%), octadecylsilane (ODS), Rhodamine 6G (R6G), sodium chloride (>99%), and methylbenzene were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents were of analytical grade and were purchased from the Aladdin Reagent Co., Ltd. (Shanghai, China). All solutions were prepared with 18 MΩ·cm deionized water acquired from a Milli-Q Water System (Billerica, MA, USA).

All chemicals were used without further purifications. Cobalt chloride hexahydrate (CoCl₂·6H₂O, 98%, M_w = 237.93), sodium laurate (C₁₁H₂₃COONa, ≥99%, M_w = 222.30), oleylamine (OAm, 70%, M_w = 267.49), oleic acid (OA, 90%, M_w = 282.46), 2-butanediol (BEG, 98%, M_w = 90.12), Ruthenium chloride (III) hydrate (RuCl₃·xH₂O, 99.98%, M_w = 207.43), hexadecylamine (HDA, 98%, M_w = 241.46), hexane (≥99%, M_w = 86.18), methylbenzene (=99.8%, M_w = 82.14), and chloroform (≥99.8%, M_w = 119.38) were purchased from Sigma-Aldrich, St. Louis, MO, USA.