Fuji dri chem slide

Blood samples were collected for ex vivo analysis. A 24-gauge catheter for invasive blood sampling collection was inserted into the ventral tail artery. After sampling arterial blood in the pre-ischemic state, heparinized physiological saline was flushed into the catheter. The catheter was kept in place until the end of the experiment. Each blood sample (0.25 mL) was obtained and placed into the i-STAT CG4 + cartridge (Abbott product, USA) and FUJI DRI-CHEM slides (Fujifilm Medical, Tokyo, Japan) of the blood test kit. Blood samples were obtained in the pre-ischemic and ischemic states (60 min and 120 min), and after pressure removal (5 min, 60 min, and 90 min). It was assessed at each time point for measurement of the inflammatory biomarker CPK.
Blood samples were analyzed using an automatic biochemical analyzer (FUJI DRI-CHEM 3500v, Fujifilm Medical, Tokyo, Japan) and handheld blood analyzer i-STAT 1 (Abbott product, Princeton, NJ, USA). Results were obtained after 2 min. The analysis included pH and CPK [13 (link),49 (link)]. Because the blood analyzer cannot display values over 2000 U/L, we decided to declare results that exceeded the upper boundary as 2000 U/L.

The serum samples were diluted 1:10 in water. The quantification of serum aminotransferases (AST) was determined by using commercially available GOT/AST Fuji DRI-CHEM slides in Fujifilm in a DRI-CHEM NX500 analyzer. The limit of detection for AST was 10 U/L.

Mice were euthanized on day 14 by intraperitoneal injection of 0.3 mg/kg medetomidine hydrochloride, 4 mg/kg midazolam, and 5 mg/kg butorphanol tartrate. Blood samples were collected from the inferior vena cava, aliquoted for later analysis, and stored at −80 °C. Immediately after blood collection, 50 mL of ice-cold phosphate-buffered saline (pH of 7.4) was slowly perfused to harvest both kidneys. Kidney samples were snap-frozen in liquid nitrogen and stored at −80 °C. Serum was separated by centrifugation at 2000 × g for 10 min. Serum urea nitrogen and creatinine concentrations were determined using a dry-chemistry system (Fuji DRI-CHEM 7000VZ: Fujifilm Corp., Tokyo, Japan) and Fuji DRI-CHEM slides (Fujifilm Corp.).

Whole animal or human blood samples were separated by being allowed to sit for 30 min or using EDTA-coated tubes, followed by centrifugation at 4000 ×g for 15 min and storage at -80°C until use. Blood glucose, triglycerides, total cholesterol, glucose, and AST/ALT were measured using Fuji Dri-Chem Slides (Fujifilm). Leptin, ghrelin, GLP-1, adiponectin, and glucagon concentrations were determined in duplicate according to the manufacturer’s ELISA instructions. ELISA kits for mouse adiponectin and leptin were purchased from Abcam (ab108785 and ab100718; Cambridge, UK), and ELISA kits for mouse ghrelin, GLP-1, and glucagon were from Cusabio (CSB-E09817m, CSB-E08118m, and CSB-E15775m, respectively; Houston, TX). Enzyme immunoassay (EIA) kits for human/mouse apelin-13 were purchased from Phoenix Pharmaceutical (EK-057-19, Burlingame, CA). ELISA kits for glucose (Ab6533, Abcam), triglycerides (ETGA-200, Bioassay Systems, Hayward, CA, USA), and cholesterol (ab65390, Abcam) were used to measure levels in the liver tissues and sera saved after the experimental diets and from control and NAFLD patients.