A drop of blood from the tail was applied to a test strip and the glucose level was measured with a Freestyle lite glucose meter (Abbot).
Freestyle lite glucose meter
Manufactured by Abbott
Sourced in United States
The FreeStyle Lite glucose meter is a compact, handheld device designed to measure blood glucose levels. It uses a small drop of blood to provide accurate and reliable readings. The device features a straightforward interface and is intended for self-monitoring of blood glucose levels.
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6 protocols using freestyle lite glucose meter
1
Measuring Glucose from Tail Blood
2
Zebrafish Fasting Glucose Measurement
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Adult zebrafish were fasted overnight and euthanized on ice for measurements under fasting conditions. Blood glucose was measured according to previously published methods [37 (link)]. In brief, blood from a pectoral girdle incision was directly applied to a Freestyle Lite glucose test strip, and glucose readings were obtained from the Freestyle Lite glucose meter (Abbott, Chicago, IL, USA) as per manufacturer’s instructions. The detection range of the instrument is between 20 mg/dL (LO) and 500 mg/dL (HI). Therefore, LO readings were recorded as 20 and HI readings as 500 mg/dL. We could assure sustained high glucose levels in analyzed pdx1−/− mutant zebrafish from 3 to 10 months of age (Figure S1 ).
3
Continuous Glucose Monitoring in Glycemia Assessment
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The Medtronic Enlite sensor (Medtronic, Northridge, California, USA) was used to monitor glycemia for two 6-day periods, separated by 2 weeks. With this sensor, interstitial glucose levels were recorded every 5 min, with a detection range of 40–400 mg/dL. Each participant was also provided with a Freestyle Lite glucose meter (Abbott, Alameda, California, USA) for self-monitored blood glucose at least twice a day to calibrate the CGM. Study physicians evaluated each CGM report, excluding periods of time with evidence of CGM malfunction or marked dyssynchrony (>30%) between CGM and fingerstick glucose values.
Glycemia metrics evaluated included measurements of overall glycemia and glucose variability over the 12-day period. TIR, glucose 70–180 mg/dL, was computed by calculating the percentage of CGM glucose readings in target range.20 (link) Time above range (TAR, glucose >180 mg/dL) was calculated in a similar manner. GMI, which is the estimation of HbA1c based on CGM data, was calculated based on mean CGM glucose over the 12 days using the previously published equation.20 21 (link) Per cent coefficient of variation was calculated as the SD divided by mean multiplied by 100 of all CGM glucose concentrations measured every 5 min over 12 days.
Glycemia metrics evaluated included measurements of overall glycemia and glucose variability over the 12-day period. TIR, glucose 70–180 mg/dL, was computed by calculating the percentage of CGM glucose readings in target range.20 (link) Time above range (TAR, glucose >180 mg/dL) was calculated in a similar manner. GMI, which is the estimation of HbA1c based on CGM data, was calculated based on mean CGM glucose over the 12 days using the previously published equation.20 21 (link) Per cent coefficient of variation was calculated as the SD divided by mean multiplied by 100 of all CGM glucose concentrations measured every 5 min over 12 days.
4
Anti-β3 Antibody in Diabetic Rats
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All rats were fasted for 4 hours; then 15 were given a single intraperitoneal injection of vehicle (0.1 M sodium-citrate buffer, pH 4.5) and 30 were given streptozotocin (STZ; 50 mg/kg) in vehicle. Hyperglycemia was confirmed one week later using tail vein blood and a FreeStyle Lite glucose meter (Abbott Laboratories, Abbott Park, IL, USA) in the STZ-treated animals. At that point daily injections of insulin (Novolin N NPH, Novo Nordisk A/S, Bagsvaerd, Denmark) of 8 units/kg were commenced (intraperitoneal injection) and continued throughout the study. The rats were maintained in a diabetic state for 4 weeks before treatment was initiated. They were then assigned to one of 2 treatment groups. One group (N = 15) received saline and one (N = 15) received the anti-β3 (C-loop) antibody intraperitoneally 0.5 mg/kg every 72 hr for 8 weeks. Urine was collected after 4 and 8 weeks of treatment. Urine was collected by placing the rats in a clean cage covered with a fresh sheet of saran wrap. The rat was left in the cage until he urinated. Urine was collected using a sterile pipette tip and placed on ice.
Rats were weighed weekly throughout each study at the same time on each occasion.
Rats were weighed weekly throughout each study at the same time on each occasion.
5
Glucose Measurement in Zebrafish
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Glucose measurements were performed using the Amplex Red Glucose Assay Kit (Invitrogen), according to manufacturer’s instructions. Larval extracts were prepared by sonication in 200 ul of cold PBS. Single 5-week old fish were first homogenized followed by sonication in 500 ul cold PBS. Extracts were spun for 15 min, 12000 rpm at 4 °C to remove debris, and the supernatant was used immediately or stored aliquoted at –80 °C until the assay was performed. Standard curves were generated with every assay. Graphs represent data combined from at least 2 independent experiments. Glucose values were normalized to protein concentration of the extract, and are reported relative to the average control value. To measure blood glucose levels in adult zebrafish, fish were anesthetized and blood collected essentially as described45 (link). Blood from the incision was directly applied to a Freestyle Lite glucose test strip, and glucose readings were obtained from the Freestyle Lite glucose meter (Abbott) as per manufacturer’s instructions.
6
Metabolic Biomarker Analysis Protocol
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The aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC) and TG levels in serum samples were measured using an enzymatic-colorimetric method and a BS-400 automatic biochemical analyzer (Mindray, Shenzhen, China), and FFA levels were measured using an RX Daytona kit (Randox, Antrim, UK). The serum levels of insulin and leptin were examined using the LINCOplex mouse serum adipokine kit (LINCO Research, Inc., St Charles, MO, USA), and serum glucose levels were measured using the Free Style Lite glucose meter (Abbott, Alameda, CA, USA). Insulin resistance (IR) was calculated using the homeostasis model assessment (HOMA)-IR with the following formula: (fasting serum glucose × fasting serum insulin)/22.5.16 (link)
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