Lx20pro analyzer

Lipid profile-related parameters to be measured include plasma total cholesterol, triglycerides, low-density and high-density lipoprotein cholesterol using a standard commercial enzymatic assay, a Beckman Coulter LX20PRO analyzer and SYNCHRON Systems lipid and multi calibrators (Beckman Coulter Diagnostics, Australia).

After 10 weeks of study (18 weeks of age), in the light phase animals were weighed and anesthetized by an i.p. injection of sodium pentobarbitone (100 mg/kg body weight; Euthatal, Delvet Limited, Seven Hills, NSW, Australia). Blood was collected from the left ventricle, centrifuged and plasma and remaining blood cells were stored at -20°C and at 4°C, respectively. Erythrocytes were analyzed for glycated hemoglobin levels (Cobas b101, Roche Diagnostics, Mannheim, Germany). Plasma was assessed for lipid and glucose levels (Beckman Coulter LX20PRO Analyzer, Beckman Coulter Diagnostics, Mount Waverley, VIC, Australia) (Watson et al., 2012 (link)). Organs were rapidly dissected and weighed (wet weight) and kidneys were either snap frozen in liquid nitrogen and stored at -80°C, or stored in buffered formalin (10%, vol./vol.) before being embedded in paraffin. With the exception of BP, experiments were conducted in a blinded fashion. A timeline for procedures and measurements is provided (Supplementary Figure 1).