Blood samples were collected in Vacuette® lithium-heparin tubes (Greiner Bio-one, Austria), centrifuged at +4°C and 2600 g for 10 min and aliquoted in polypropylene Nunc Cryotube™ vials (Thermo Scientific, Denmark) within 60 min from collection. MD samples were collected in microvials (M Dialysis) and stored at approximately −20°C. At the end of each study day all samples were transferred to a −80°C freezer until further analysis.
Vacuette lithium heparin tubes
Manufactured by Greiner
Sourced in Austria, Germany
Vacuette® lithium-heparin tubes are used for the collection and transportation of blood samples. They contain lithium heparin as an anticoagulant to prevent the blood from clotting.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll paque plus, by GE Healthcare (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Nunc cryotube vial, by Thermo Fisher Scientific (1 mentions)
Neutral buffered formalin, by Merck Group (1 mentions)
Cyclophosphamide, by Merck Group (1 mentions)
Fluorochrome conjugated mabs, by BD (1 mentions)
Zombie nir live dead stain, by BioLegend (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
5 protocols using vacuette lithium heparin tubes
1
Blood Sample Handling Protocol
2
Isolation and Phenotyping of Human Immune Cells
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VACUETTE lithium heparin tubes were obtained from Greiner Bio-One (Frickenhausen, Germany). Ficoll-Paque PLUS was obtained from GE Healthcare (Uppsala, Sweden). PBS, RPMI-1640 medium, GlutaMAX, and FCS (heat-inactivated at 56 °C for 30 min) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Sterile isotonic saline was obtained from Pfizer (Sydney, NSW, Australia). BBG, cyclophosphamide, and 10% neutral-buffered formalin were obtained from Sigma-Aldrich (St Louis, MO, USA). Sterile BBG (5 mg/mL) was prepared in isotonic saline, and sterile cyclophosphamide (6.6 mg/mL) was prepared in PBS. Haematoxylin and eosin stain was obtained from POCD Scientific (Artarmon, NSW, Australia). Zombie NIR live/dead stain was obtained from BioLegend (San Diego, CA, USA). Fluorochrome-conjugated mAbs, as described in detail [24 (link)], and R-phycoerythin-conjugated anti-human CD33 mAb (clone P67.6) were obtained from BD Biosciences (San Diego, CA, USA).
3
Porcine Blood Sampling and Analysis
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Blood samples were collected from all pigs (i.e. N = 16 per diet) at the beginning of the feeding trial (day 0) and after 16 and 77 days, and from all pigs in the 9-pig pens at 49–56 days into the trial (0 h) and 3, 5, 8, 12, 24 and 48 h after the LPS injection. Pigs were immobilized using a rope on upper jaw and blood samples were collected via the vena jugularis externa using 6-mL Vacuette Lithium Heparin tubes (Greiner Bio-One, Austria) with Venoject needles (20G × 1½’’UTW, USA). Lithium heparin blood was subjected to haematological analysis, whereas heparinized plasma was isolated by centrifugation at 3500 g for 15 min (Megafuge 1.0 R, Heraeus SEPATECH, USA) for subsequent clinical biochemistry, Se and VitE analyses. Samples were stored at – 20 °C or – 70 °C until the day of analysis.
4
Pharmacokinetic Profiling of Antiviral Compound
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All blood and urine collection was taken on day 5 of each treatment session. Venous blood samples were collected at pre-dose (0 h) and at 0.5, 1, 2, 3, 4, 5, 6, 8, 10, and 12 h post-dose. Blood samples were collected from a catheter, which was placed in the forearm vein before dosing. At the specified time, 5 mL of blood was drawn (except for 0 time and at 2 h which was 10 mL each for additional antiviral activity determination) and stored in vacuette lithium heparin tubes (Greiner Bio-One). Dichlorvos (a carboxylase inhibitor) was then added (200 μg/mL) into the blood samples (except for those used for antiviral activity determination) to prevent in vitro hydrolysis from OA to OC [8 (link)–10 (link)]. Plasma samples were collected after centrifugation and then stored at −80°C until assay. Urine was also collected (and the volume was recorded) at pre-dose, 0–4, 4–8, and 8–12 h intervals. The urine samples (~10 mL) were stored at −80°C until assay.
5
Isolation of human PBMCs
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All experiments with human blood were conducted in accordance with approval by the Human Ethics Committee, University of Wollongong. Peripheral blood was collected by venepuncture into VACUETTE® lithium heparin tubes (Greiner Bio-One; Frickenhausen, Germany). Whole blood, diluted with an equal volume of sterile phosphate buffered saline (PBS) (Thermo Fisher Scientific, Waltham, USA), was underlaid with Ficoll-Paque PLUS (GE Healthcare; Uppsala, Sweden) and centrifuged (560 x g for 30 min). hPBMCs were collected and washed with PBS (430 x g for 5 min) and resuspended in PBS.
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