At the end of the HFD feeding period, the animals were deprived of food for 2–5.5 h from the start of the light phase, before being killed; decapitation was in a random order. Blood glucose concentrations were measured using a standardized method (Freestyle). MiniCollect serum tubes (Greiner Bio-One BV) were used to collect serum. Livers were collected, weighed, and snap-frozen in liquid nitrogen. Gonadal WAT (gWAT) was excised; left fat pads were snap-frozen in liquid nitrogen, whereas right fat pads were weighed and fixed in paraformaldehyde. Subcutaneous WAT (sWAT) was collected from the inguinal region and snap-frozen in liquid nitrogen. Pancreatic tissue was separated from mesenteric WAT, based on density in PBS; both organs were weighed and pancreatic tissue was fixated in paraformaldehyde. Hypothalami were collected and snap-frozen in liquid nitrogen. All samples were stored at −80°C until analysis.
Minicollect serum tubes
Manufactured by Greiner
Sourced in Germany, United States
MiniCollect serum tubes are a type of lab equipment designed for the collection and transportation of blood samples. They are used to obtain serum samples from patients for various laboratory tests and analyses.
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Lab products found in correlation
8 protocols using minicollect serum tubes
1
Metabolic and Tissue Analysis after HFD
2
Rodent Tissue Sampling and Analysis
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At the end of PW 6 and PW 15, mice were deprived of food at the onset of the light phase and decapitated two to six hours thereafter. Blood glucose was measured in duplicate with a Freestyle glucose meter (Abbott Diabetes Care, Hoofddorp, The Netherlands). Whole blood was collected in chilled MiniCollect serum tubes (Greiner Bio-One BV, Alphen aan de Rijn, The Netherlands), spun down at 4 °C for 10 min at 3000× g, and the resulting serum aliquoted and stored at −80 °C. Liver, mesenteric white adipose tissue (mWAT), and pancreas were weighed and snap-frozen in liquid nitrogen. A ~2 g clip was attached to the distal end of the small intestine and hung next to a ruler to determine the length of the small intestine. Thereafter, the small intestine and colon were each cut longitudinally, rinsed in ice-cold RNase-free phosphate-buffered saline to remove their contents, and weighed separately. Caecum contents were extracted, weighed, and snap-frozen. One pad of gonadal white adipose tissue (gWAT) was snap-frozen; the other pad was weighed, fixated in 4% paraformaldehyde overnight, and embedded in paraffin. Samples were stored at −80 °C until further analysis.
3
Ferret Tissue Sampling for Virus Analysis
4
Serum Enzyme Activity Quantification
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Blood was collected from the inferior vena cava into MiniCollect Serum Tubes (Greiner Bio-One, Kremsmünster, Germany). Serum was isolated by centrifugation at 3000 relative centrifugal force (RCF) for 10 minutes and the supernatant was collected. Serum was diluted 1:3 in PBS and tested for the activity of enzymes alkaline phosphatase (ALP), alanine transaminase (ALT) and aspartate transaminase (AST) by the Sydney South West Pathology Service. All results are measured in international units per litre (U/L).
5
Ferret Tissue Sampling and Analysis
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On sampling days, blood was collected and placed in MiniCollect EDTA tubes (Greiner Bio-One, Monroe, NC) for virus load and hematology analysis or MiniCollect serum tubes (Greiner Bio-One) for clinical chemistry and antibody analysis. Necropsy was performed on all ferrets and tissues sampled included lungs, liver, spleen, kidney, adrenal gland, pancreas, and brain (frontal cortex). Ten percent tissue homogenates of liver, spleen, kidney, adrenal gland, and brain were used for virus load analysis.
6
Blood and Adipose Tissue Sampling
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Food was removed 2 h before the mice were killed in the morning (light phase). Mice were directly killed by decapitation, and blood was partially collected in 60 µL heparinised capillary tube (Hirschmann Laborgeräte, Eberstadt, Germany) for determination of whole blood haematocrit levels. Capillary tubes were centrifuged in a micro-haematocrit centrifuge at 3000× g for 5 min. Haemoglobin (Hb) levels were measured using an automated Hb monitoring system (Hemocue 201 Plus, HemoCue Ltd., Angelholm, Sweden) with 10 µL micro cuvettes. Remaining blood was collected via a funnel into mini collect serum tubes (Greiner Bio-one, Longwood, FL, USA) and spun down 10 min at 5780× g at 4 °C. Epididymal WAT (eWAT) was excised rapidly, left portion weighed and snap frozen in liquid nitrogen. The right eWAT portion was fixed overnight in 4% paraformaldehyde, washed with PBS and paraffin embedded for histological analysis. We focused on eWAT because it is the most widely examined WAT depot in mice, it is a visceral AT depot and visceral AT is most strongly associated with disease risk, and eWAT responds first to an obesogenic condition.
7
Glucose Metabolism in Fasted and Fed Mice
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Mice were food restricted (1.1 g HFD) for 16 h, starting at ZT = 11. Half of the mice was sacrificed in the post-absorptive state, and the other half was administered the liquid mixed meal with 13C glucose by oral gavage and sacrificed after 45 min (postprandial state) by decapitation. Trunk blood was collected in MiniCollect serum tubes (Greiner Bio-One, Alphen aan de Rijn, The Netherlands). Serum was separated by centrifugation at 4 °C for 10 min at 3000 × g, aliquoted, and stored at −80 °C. Glucose was measured in whole blood with a Freestyle glucose meter (Abbott Diabetes Care, Hoofddorp, The Netherlands) directly after sacrifice.
8
Glucose Challenge Test in Mice
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On day 105, mice were food-deprived at the start of the light phase for 2–4 h. Fifteen minutes before sacrifice, mice were given a glucose bolus (2.0 g/kg body weight) by oral gavage, to have them in a challenged state with peaking insulin levels, upon sacrifice. Mice were decapitated, and whole blood glucose levels were immediately measured (Freestyle, Abbott Diabetes Care, Hoofddorp, The Netherlands). The remainder of the blood was collected in MiniCollect serum tubes (Greiner Bio one B.V., Alphen aan de Rijn, The Netherlands), centrifuged for 10 min at 3000× g at 4 °C, and the obtained serum was aliquoted and stored at −80 °C. Liver, gWAT depot, and skeletal muscle (extensor digitorum longus muscle) tissue were collected, snap-frozen in liquid nitrogen, and stored at −80 °C until further analysis.
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