Mayer s hematoxylin solution

In this study, a standard laboratory procedure called H&E staining was performed on 4.0 μm serial sections of the samples. To begin the staining process, the paraffin‐embedded tissue sections were deparaffinized, which involves removing the paraffin wax from the sections. Next, the sections were subjected to H&E‐stained, which involves the use of Mayer's hematoxylin solution (Solarbio, Beijing, China). The staining procedure was carried out according to the instructions provided by the manufacturer of the hematoxylin solution. Following the staining process, the microscope system from LEICA, a renowned German manufacturer, was utilized to capture images of the stained sections.

DA5‐CH had been synthesized by China Peptides. It had a purity of 95%. Its purity was confirmed by reversed‐phase high‐performance liquid chromatography (HPLC) and characterized by matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) mass spectrometry. The drug was stored at −20°C and dissolved in saline before intraperitoneal injection in rats. Streptozotocin (STZ) was bought from Sigma‐Aldrich. It was stored at −20°C and dissolved in artificial cerebrospinal fluid before lateral ventricular injection in rats.
Rabbit anti‐tau, p‐tau (phospho ser396), synaptophysin, p‐CREB (phospho Ser133), CREB, and anti‐rabbit IgG were purchased from Abcam. Antibodies against B‐celllymphoma‐2 (Bcl‐2), Bax were bought from Boster. The antibody to PSD95 was obtained from Proteintech. BCA protein assay kit and Mayer's Hematoxylin solution were purchased from Solarbio. Sodium chloride, ethylene glycol, and 3,3‐diaminobenzidine (DAB) were obtained from ZSGB‐BIO Co.

Liver and adipose tissue were immediately excised and parts were fixed in 4% paraformaldehyde after mice were sacrificed and then embedded in paraffin. Sections of 5 μm were stained with hematoxylin and eosin (HE) (Solarbio, China). Other parts of liver were snap frozen with liquid nitrogen and then cryostat-sectioned at a thickness of 10 μm onto glass slides for oil red O staining. These sections were fixed with 4% paraformaldehyde and briefly washed with running tap water. Then, sections were rinsed with 60% isopropanol for 1 min, followed by staining with freshly prepared oil red O working solution (Solarbio, China) for 15 min, and differentiated with 60% isopropanol for 1 min. Nuclei were briefly stained with Mayer’s Hematoxylin solution (Solarbio, China), followed by rinsing with distilled water. Sections were mounted with glycerol gelatin aqueous slide mounting medium (Solarbio, China). Positive staining was quantified using Image J. Adipocyte sizes were quantified from WAT histology slides and were quantified with Adiposoft software as described (20 (link)). Slides were imaged with microscope (Olympus BX-50, Olympus Optical, Japan).