Eosin staining solution

The paraffin-embedded adipose tissue was cut into small sections, dewaxed twice through xylene, and sections were sequentially soaked in alcohol at different concentrations followed by washing with distilled water. The sections were then stained with hematoxylin (Servicebio, Wuhan, China), washed in distilled water, dehydrated with 0.5% hydrochloric acid alcohol. Next, the sections were stained in an eosin staining solution (Servicebio, Wuhan, China), and dried after washing with distilled water. Lastly, neutral resin (Servicebio, Wuhan, China) was used to seal the sections, and photos were taken under a microscope (Leica DM IL LED Fluo, Germany).

The kidney tissue was fixed in 4% paraformaldehyde. Slices of the paraffin-coated kidney samples were taken. 5 μm paraffin sections were dewaxed using a seines of ethanol. The sections were subjected to sequential immersion in xylene I/II, absolute ethanol I/II, and 75% alcohol for durations of 20, 5, and 5 min, respectively, and subsequently rinsed with water. Subsequently, the sections were subjected to hematoxylin staining (Servicebio, Wuhan, China) for a duration of 3–5 min. Following sequential dehydration in 85% and 95% alcohol, the sections were subjected to staining with an eosin staining solution (Servicebio, Wuhan, China) for a duration of 5 min. Subsequently, the process of dehydration and sealing was executed, followed by the acquisition of images utilizing a light microscope. Scores on the HE staining for interstitial dilatation and inflammatory cell infiltration ranged from 0 to 5. 0: normal; 1: <10% positive; 2: 10–25% of tubules injured area positive; 3: 25–50% positive; 4: 50–75% positive; 5: >75% positive [24 (link)].