We measured RBC deformability and aggregation to evaluate RBC hemorheological function. Uyuklu et al. [34 (link)] recommended that RBC deformability and aggregation should be analyzed at 25 °C at shear stress of 3 Pa within 4–6 h after collecting blood, so all samples were analyzed within 30 min of their collection at a room temperature of 25 °C using a Rheoscan-D (Rheo Meditech Inc., Seoul, Korea). For RBC EI analysis, the sample was transferred to a 2 mL microfuge tube and then diluted in 700 μL of 5.5% polyvinylpyrrolidone (360 kDa) dissolved in 1 mmol phosphate buffered saline (pH 7.4; osmolality = 300 mOsmol/kg) in a K3EDTAtube (Greiner bio-one, Chon Nuri, Thailand). Then, 0.5 mL of this solution was analyzed using a D-test kit according to manufacturer’s instructions (Rheo Meditech Inc.). For the accuracy of RBC EI measurement, a Lineweaver-Bruke plot model (LB model) was used [35 (link)]. For the RBC AI analysis, 8 μL of the blood sample (direct whole blood analysis) was analyzed using an A-test kit according to manufacturer’s instructions (Rheo Meditech Inc., Seoul, Korea).
K3edta tube
Manufactured by Greiner
Sourced in Thailand, Austria, United Kingdom
The K3EDTA tube is a laboratory specimen collection device used to gather blood samples. It contains the anticoagulant K3EDTA, which helps prevent the blood from clotting. This allows the collected sample to be used for various laboratory analyses.
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Lab products found in correlation
4 protocols using k3edta tube
1
Evaluation of Red Blood Cell Rheology
2
Optimizing Blood Collection for Liquid Biopsy
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Prostate cancer patients (n = 3) and healthy female blood donors (n = 4) provided written informed consent to participate in the study. The study was approved by the South Western Sydney Local Healthy District Ethics Committee, Australia (HREC/13/LPOOL/158; 02/09/2013). Peripheral blood was drawn by venipuncture into blood tubes with the initial 3 mL discarded to prevent keratinocyte contamination and false positive CTCs being present. For the main comparison experiment, a total of 192 mL blood from each healthy donor (n = 3) was drawn into a total of 24 blood tubes including: 4 × 9 mL K3EDTA tube (Greiner Bio-One, Kremsmünster, Austria), 4 × 9 mL acid citrate dextrose-B tube (Greiner Bio-One, Kremsmünster, Austria), 4 × 10 mL Cell-free DNA BCT (Streck, Omaha, NE, USA), 4 × 10 mL Cell-free RNA BCT (Streck, Omaha, NE, USA), and 4 × 2 × 5 mL Cyto-Chex BCT (Streck, Omaha, NE, USA). In the follow-up experiment to test increased proteinase K treatment, 38 mL blood from two healthy donors was drawn into one set of EDTA, Citrate, and DNA BCT and RNA BCT tubes. In the experiment with prostate cancer patients, blood was drawn into 3 × 6 mL K2EDTA tubes (BD, Franklin Lakes, NJ, USA) available in the clinic.
3
Multiomics Sample Preparation Protocol
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Plasma, serum and urinary samples were collected using the K3EDTA Tube (Greiner Bio-One Ltd, Stonehouse, UK), the BD Vacutainer® SSTTM II Advance Serum Tube (BD, Plymouth, UK) and the SterilinTM Polypropylene 30 mL Universal Container (Thermo Scientific, Wilmington, DE, USA), respectively. Plasma and serum samples were immediately isolated by centrifugation at 1500 x g for 15 min at 4˚C, before storage at -80°C for further processing. Three 1 mL urine aliquots per sample were stored at -80°C prior to downstream analysis. A volume of 100 µL isolated plasma, serum or urine was each mixed vigorously with 100 µL chloroform and 300 µL methanol on a cooled shaker at 4˚C for 1 hr, followed by centrifugation at 13,000 x g at 4°C for 3 min. The supernatant was transferred to fresh screw capped tubes stored at -80°C until transportation packed with dry ice to the Polyomics Facility, University of Glasgow, for the hydrophilic interaction liquid chromatography (HILIC)-mass spectrometry (MS).
4
Standardized Plasma Collection and Storage
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Samples were collected in EDTA tubes using a 23 gauge butterfly needle and vacutainer needle holder following standard venepuncture in the cubital region, in the area of anastomosis between the radial and the ulnar veins or in the brachial vein, slightly proximal to this area. To avoid coagulation of the sample, a 9ml K3EDTA tube (Greiner Bio-One, Cat # 455036X -EDTA tube, lavender lid) was used. The sample was immediately kept at +4°C until being processed within two hours. The tube was labelled with the subject barcode with no phenotypic information given and then centrifuged at 3000 rcf (xg), 8 minutes at +4°C; the plasma was aliquoted in 0.5 ml eppendorf tubes and frozen at -80°C within three hours of collection.
Analysis of results for combined plasma and serum samples (n=194), and with serum samples alone are presented (n=134).
Analysis of results for combined plasma and serum samples (n=194), and with serum samples alone are presented (n=134).
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