Dm2500 optical microscope

Lung tissue was fixed in a 4% paraformaldehyde followed by paraffin embedding. The tissue wax blocks were cut into 5 μm slices by a tissue slicer. Following this, the sections were stained with hematoxylin and eosin (H&E). The 5 μm-thick tumor tissue slides were treated with 3.0% hydrogen peroxide and closed with sheep serum for immunohistochemical analysis. They were incubated with the CD206 primary antibody (1:400 dilution, Abcam, Cambridge, MA, USA) at 4 °C overnight, followed by incubation with horseradish peroxidase (HRP)-conjugated secondary antibody (goat anti-mouse IgG, CWBIO, Taizhou, China) at room temperature and detection with diaminobenzidine with hematoxylin as a reverse stain. Photography was performed using the LEICA DM2500 Optical Microscope.

Anemoside B4 (B4), whose chemical structure is shown in Figure 1(a) (Batch No. 20161107, purity of 98% using HPLC determination, the chromatogram of anemoside B4 is shown in Figure 1(b)) was presented by Professor Yu-Lin Feng from the Phytochemical Department of our university. Prednisolone (pred.) (11-beta,17-alpha; 11,17,21-trihydroxypregna-1,4-diene-3,20-dione; (11beta)-11,17-dihydroxy- 3,20- dioxopregna-1,4-dien-21-yl acetate) (Batch No. 161165) was purchased from Xianju Pharmaceutical Ltd. (Zhejiang, China). Adenine (Sigma-Aldrich, Lot#WXBB0585V) was purchased from Sigma (Shanghai, China). Creatinine (Crea), urea nitrogen (BUN), and total protein (TP) kits were purchased from Heguan Chemical Company (Beijing, China). The Bradford Protein Assay Kit (P0006) was purchased from Beyotime Biotechnology Company (Beijing, China). The qPCR detection kit (Lot# L20509) was purchased from Quanshijin BioChem Tech Ltd. (Beijing, China). Hematoxylin (MHS16-500ML, LOT#SLBK4909V) and Eosin Y (HT110116-500ML, LOT#J6425V) were purchased from Sigma (Shanghai, China). Masson's trichrome kits (Lot:0606A18) were purchased from Leagene Biotechnology Co., Ltd. (Beijing, China). A Bio-Rad electrophoresis unit and Bio-Rad ChemiDocXRS+ Gel Imaging System (Beijing, China), LEICA RM2235 paraffin slicer, and LEICA DM2500 Optical Microscope (Beijing, China) were used in this study.

The thickness of polymer thin films and polymer-pixels were measured through a profilometer (Bruker Dektak XT Stylus Profilometer). Atomic Force Microscope (AFM—NT-MDT Solver NEXT equipped with an S7 scanner with a scan area of 5 µm by 5 µm) was used for the morphology investigation of polymer rounded pixels. A minimum of three separate scans were recorded for each sample. The AFM data were analysed with SPM software Gwyddion v2.51. Optical microscope (Leica DM 2500 optical microscope) was used for the morphological characterization of the pixelated artificial retina device. Confocal microscope (Olympus Lext—3100) was used for aged samples morphology analysis. All measurements were performed at ambient conditions.

Raman spectra were performed on M-GLY, COT and COT/M-GLY residues by using a Renishaw InViaTM Raman (Gloucestershire, UK) equipped with an argon laser (514 nm wavelength, 50 mW power, 3 scans) coupled with a Leica (Wetzlar, Germania) DM 2500 optical microscope.