Food intake was measured for mice placed in individual cages during at least 3 consecutive days. Lean mass, fat mass, and percent fat were determined on awake mice using an Echo MRI analyzer (Whole Body Composition Analyzer, Echo medical systems, Houston, TX, USA). Females underwent a glucose tolerance test (GTT) after an overnight fast. Glucose (2 mg/g b.w.) was administered intraperitoneally, and glucose levels were determined in tail blood using an ACCU‐CHEK® Aviva blood glucose monitor and appropriate test strip (Roche Diagnostics). Oxygen consumption (VO2), carbon dioxide production (VCO2), and spontaneous locomotor activity were determined using indirect, open‐circuit calorimetry in an Oxymax Metabolic Chamber system (Columbus Instruments, Columbus, OH, USA). The mice (with free access to food and water) were housed in separate chambers for 48 h for acclimatization before starting 48 h measurements. The average VO2 was calculated during 48 h—two light cycles and two dark cycles (Somm et al, 2005 , 2014 ).
Accu chek aviva blood glucose monitor
Manufactured by Roche
Sourced in United Kingdom, Switzerland
The Accu-Chek Aviva is a blood glucose monitor designed to measure blood glucose levels. It is a compact, handheld device that requires a small drop of blood from the fingertip to provide a reading.
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5 protocols using accu chek aviva blood glucose monitor
1
Metabolic Phenotyping of Mice
2
Intraperitoneal Glucose Tolerance Test in Mice
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An intraperitoneal glucose tolerance test (IPGTT) was carried out on 8 mice from each diet group after 8 weeks of experimental diet feeding. Mice were weighed and then fasted for 5 hr from 8:00 a.m. Glucose was prepared as 100 mg/mL in saline, filter sterilized, and stored at −20°C. An initial blood sample was taken at 0 min via a tail-tip cut removing 1 mm of tail tip prior to the intraperitoneal glucose injection containing 1.5 mg glucose per gram of body weight. Subsequently, blood samples were taken from the tail vein at 15, 30, 60, and 120 min by gently squeezing the tail to remove the scab and express a fresh spot of blood. The first spot of blood was discarded, and the second spot of blood was transferred directly from the tail onto an Accu-Chek Aviva Test Strip (Roche Diagnostics, Burgess Hill, UK). Blood glucose was measured using an Accu-Chek Aviva blood glucose monitor (Roche Diagnostics, UK). Areas under the curve for individual IPGTTs were calculated using SigmaPlot 13.0 (Systat Software, Hounslow, London, UK).
3
Insulin Sensitivity Assay in Mice
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After 8 weeks on the diets, mice were fasted for 4–5 h in the middle of the dark phase with ad libitum access to water. Actrapid HM human insulin (Novo Nordisk, Denmark) was injected intraperitoneally (IP), and tail blood glucose levels were monitored at the time points indicated using the Accu-Chek Aviva blood glucose monitor (Roche, Switzerland). The Insulin dose was 0.4 and 0.8 mU/g body weight for CD and HFD fed mice, respectively.
4
Fasted Mouse Insulin Tolerance Protocol
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Mice were fasted for 5–6 h in the middle of the dark phase with ad libitum access to water. Actrapid HM human insulin (Novo Nordisk) was injected intraperitoneally (IP), and tail blood glucose was monitored at the indicated time points using the Accu-Chek Aviva blood glucose monitor (Roche). Insulin dose: 0.4 mU/g body weight (CD) and 0.8 mU/g body weight (HFD)19 (link).
5
Hyperinsulinemic-hypoglycemic clamp in mice
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The hypoglycemic clamp was performed following 5 h of fasting. Surgery and clamp experiments were performed as previously described (17) with the protocol modification of returning of red blood cells (18) . Briefly, the right jugular vein and the left carotid artery were catheterized using catheters filled with heparinized saline (100 U/mL). The mice remained anesthetized to reduce variation in the blood glucose concentrations due to stress. Following baseline sampling, synthetic human insulin (Actrapid®, Novo Nordisk, Bagsvaerd, Denmark) was infused as a continuous infusion (15 mU/kg animal/min) at a pace of 2 μL/min for 90 minutes.
Blood glucose in ~5 μL whole blood was determined every 10 minutes with an Accu-Chek Aviva blood glucose monitor (Hoffman-LaRoche). A variable amount of a 10% glucose (Sigma-Aldrich, MO, USA) solution was infused to maintain blood glucose levels at 2.5 mmol/L. Glucose requirement to maintain target glucose was represented by the glucose infusion rate (GIR) during the final 30 min steady state of the clamp.
Blood glucose in ~5 μL whole blood was determined every 10 minutes with an Accu-Chek Aviva blood glucose monitor (Hoffman-LaRoche). A variable amount of a 10% glucose (Sigma-Aldrich, MO, USA) solution was infused to maintain blood glucose levels at 2.5 mmol/L. Glucose requirement to maintain target glucose was represented by the glucose infusion rate (GIR) during the final 30 min steady state of the clamp.
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