H e staining kit

The specimens were fixed with 4% paraformaldehyde in 0.1 M phosphate buffer, at pH 7.4 overnight at 4°C. The tissues were dehydrated through a series graded ethanol baths (100, 95, 90, 80 and 70%), and the specimens were embedded in paraffin. The embedded tissue was cut into 4 µm sections. The center of the scanned region was set for histological evaluation.
TUNEL staining of fractured double-stranded DNA in apoptotic cells was performed. Paraffin sections (4 µm) of synovial and cartilage tissues were used for the evaluation of apoptosis by TUNEL staining. The In Situ Cell Death Detection kit-POD (Roche Diagnostics, Basel, Switzerland; cat. no. 11684817910) was used to determine the number if apoptotic cells, according to the manufacturer's protocol. Counting of TUNEL-positive cells was conducted on ×200-magnified sections for each group. A H&E Staining kit (Shanghai Yeasen Biotechnology Co., Ltd., Shanghai, China, cat. no. 60524ES60) was used for staining the cardiac muscle, according to the manufacturer's protocols. Briefly, slides were stained for 5 min with hematoxylin, washed with diluted water three to five times at room temperature and subsequently stained with eosin solution for 20 sec at room temperature.

The kidney samples were fixed using 10% formalin, paraffin-embedded, and cut into 5-µM-thick sections. The sections were then subjected to Masson's trichrome staining using a modified Masson's Trichrome stain kit (Solarbio Science & Technology Co., Ltd.) according to the manufacturer's instructions. Briefly, the sections were stained in Harris hematoxylin for 5 min, in Ponceau acid fuchsin staining solution for 10 min, and in Aniline Blue solution for 5 min at room temperature. Hematoxylin and eosin (H&E) staining was performed using a H&E staining kit (Yeasen Biotech Co., Ltd.) according to the manufacturer's instructions. Briefly, the sections were stained in hematoxylin for 5 min and in eosin solution for 1 min at room temperature. Images were obtained using an Olympus IX73 microscope (Olympus Corporation). ImageJ software for Windows V 1.52v (NIH) was used to quantify the Masson's trichrome positive staining area.