Serum and tissue levels of metabolites and electrolytes using Cobas Clinical Chemistry Analyzer
The assays will be performed using solid state chemistry available with the Vitros DT 60 system. Each requires about 10 µl of plasma. The VITROS DT60 II Analyzer and modules can perform up to 65 colorimetric tests/hr including: ammonia, amylase, cholesterol, creatinine, BUN/UREA, glucose, HDL-cholesterol, lactate, magnesium, neonatal bilirubin, phosphorous, total bilirubin, total protein, triglycerides, uric acid. The following tests can be run at a rate of 20/hr: albumin, alkaline phosphatase, ALT, AST, Ca2+, CPK, CPK-MB, creatinine, GGT, Fe, LDH, lipase, urine creatinine. The following electrolytes can be measured at a rate of 15 electrolyte tests/hr: CO2, NaCl, K+. Lastly, the following derived tests will be offered: A/G ratio, anion gap, coronary risk classification, cholesterol/HDLC ratio, globulin, LDL, VLDL. Microassays for NEFA will be performed in 96 well plates using an assay from Wako. Plasma glycerol and ß-hydroxybutyrate concentrations will be determined using enzymatic methods. Blood for ammonia levels must be kept on ice immediately and assayed immediately. Electrolytes and cations should be measured from heparinized plasma samples. We have specialized caging with Teflon-treated and humidified & chilled collection devices and collection chambers to separate urine and feces from mice. Fecal fat will be determined by weighing the extracted lipids from 1g aliquots of freeze-dried powdered feces that have been lipid extracted.