Umbilical cord blood was collected immediately after delivery in Vacutainer® Plus Plastic K2EDTA Tubes (BD, Franklin Lakes, NJ, USA). Samples were centrifuged at 3,200 rpm for 15 min to retrieve buffy coats and instantly frozen, first at −20°C and later at −80°C. Placentas were deep-frozen within ten minutes of delivery. Afterwards, we took biopsy samples of approximately 1 to 2 cm3 for DNA extraction using a standardized protocol as described by Adibi et al. [30 (link)]. Four distinct sites from nineteen placentas were sampled from the foetal side across the middle region of the placenta approximately four cm away from the umbilical cord and 1–1.5 cm below the chorio-amniotic membrane (Figure 2 ). An extra biopsy was taken from the maternal side. Chorio-amniotic membrane contamination was avoided by careful visual examination and dissection. Via histological examination (hematoxylin & eosin staining) we compared four fetal biopsies from four placentas. This confirmed that biopsies were taken from chorionic villous tissue with normal architecture composed of trophoblasts. In the fetal biopsies we could identify terminal and intermediate villi with cytotrophoblasts and syncytiotrophoblasts, vessels, fibrin and the intervillous space. We did not observe any consistent differences in the histology or cell type composition between the fetal samples, nor between the four placentas.
Vacutainer plus plastic k2edta tubes
Manufactured by BD
Sourced in United States
The BD Vacutainer® Plus Plastic K2EDTA Tubes are laboratory blood collection tubes used for the collection and transport of venous blood samples. The tubes contain the anticoagulant K2EDTA, which prevents blood clotting during the collection and transportation process.
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Lab products found in correlation
Vacutainer eclipse blood collection needle, by BD (2 mentions)
Rnaeasy column, by Qiagen (2 mentions)
Trizol ls reagent, by Thermo Fisher Scientific (2 mentions)
Cell dyn 3500, by Abbott (1 mentions)
Mestrenova, by Mestrelab Research (1 mentions)
Avance 600, by Bruker (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Chloroform, by Merck Group (1 mentions)
Vacutainer plus plastic serum tube, by BD (1 mentions)
Advia centaur cortisol assay, by Siemens (1 mentions)
Qiaamp dna mini kit, by Qiagen (1 mentions)
13 protocols using vacutainer plus plastic k2edta tubes
1
Placental Tissue Sampling for DNA Extraction
2
Blood Biomarker Profiling Protocol
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Blood samples were collected in the morning until 1100 hours, after fasting in Vacutainer® Plus Plastic K2EDTA Tubes (BD, Franklin Lakes, NJ, USA) and PAXgene Blood RNA vacutainer tubes (PreAnalytiX, Qiagen, Hilden, Germany). Blood cell counts and differential leukocyte counts were determined using an automated cell counter with flow differential (Cell Dyn 3500, Abbott Diagnostics, Abott Park, IL, USA). Blood glucose levels, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, and C-reactive protein (CRP) were measured according to standard clinical procedures.
3
PBMC Isolation from ALS Patients
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Blood was drawn by standard venipuncture into Vacutainer® Plus Plastic K2EDTA Tubes (Becton, Dickinson and Company) and kept at 4°C until shipment to the Istituto di Ricerche Farmacologiche Mario Negri IRCCS. PBMCs were isolated from ALS patients and healthy individuals, as previously described.7 (link) Further details are described in the Supplementary material .
4
PBMC Isolation from ALS Patients
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Blood was drawn by standard venipuncture into Vacutainer® Plus Plastic K2EDTA Tubes (Becton, Dickinson and Company) and kept at 4°C until shipment to the Istituto di Ricerche Farmacologiche Mario Negri IRCCS. PBMCs were isolated from ALS patients and healthy individuals, as previously described 7 (link) . Further details are described in the Supplementary material.
5
Blood Sample Collection and Plasma Extraction
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For both FM and CON, venous blood samples were collected using a Vacutainer (BD Vacutainer Eclipse Blood Collection Needle; BD Diagnostics, Becton, Dickinson, and Company, Franklin Lake, NJ) in 10 mL K2EDTA tubes (BD Vacutainer Plus Plastic K2EDTA Tubes, BD Diagnostics). The plasma was retrieved by centrifuging the blood samples for 30 min at 1500g at RT immediately after collection. The plasma fraction was removed to a new tube before aliquoted and stored in −86°C until analysis. The retrieval of the blood samples at baseline was performed after the clinical examination, within 1–7 d before the start of the intervention and within 1–7 d after the intervention.
6
Serum Metabolite Profiling by NMR Spectroscopy
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In this study, peripheral blood samples (6 mL) were collected and preserved in collection tubes (BD Vacutainer Plus plastic K2EDTA tubes) and transported to the laboratory within 5 minutes of collection. The red blood cells were removed by centrifugation at 3,000 rpm for 15 minutes to exclude erythrocytes from serum. Serum samples were then transported into cryogenic tubes and stored at −80°C. Serum samples (200 μL) were diluted in D2O (400 μL) containing 1 mM sodium salt of 3-(trimethylsilyl) propionic-2,2,3,3, d4 acid (TSP). Solvent residual peak of D2O was considered as standard in NMR experiments. Water peaks were suppressed for minimizing the noise of the NMR spectrum.1H-NMR spectra were acquired on a 600 MHz (9.4 T) magnet interfaced to a spectrometer (Avance 600; Bruker, Rheinstetten, Germany) and kept at 30°C throughout the experiment. Before assessment, each spectrum was digitized in 1,000 bins of identical widths so that all peaks were considered. Once digitized, they were fed into MestReNova (ver 9.0.0.12821; Mestrelab Research, S.L., Santiago, Spain). After checking and comparing the concentrations (in parts per million) and intensities with those in the Human Metabolome Database (HMDB),27 (link) we could enquire which NMR peaks (ie, metabolites) were responsible for the chemical shifts.
7
Blood Lipid Profile Analysis
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After eight weeks of treatment, whole blood of each mouse was collected via the retroorbital plexus to BD Vacutainer® Plus Plastic K2 EDTA tubes (BD Biosciences, San Jose, CA) and centrifuged 1,000 ×g for 20 min at 4°C. The supernatant was transferred to a fresh tube for determination. Plasma TGs, TC, LDL, and HDL levels were analyzed at the Seoul Medical Science Institute (SCL, Seoul, South Korea).
8
Isolation of Healthy Neutrophils
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Fresh peripheral venous bloods were collected from healthy volunteers with no history of candidiasis in BD Vacutainer® Plus Plastic K2EDTA Tubes or serum separator tubes for different purposes. Neutrophils were isolated as described previously (26 (link)). Briefly, bloods were diluted in Mg2+- and Ca2+-free DPBS and overlaid on Ficoll-Paque solution (Ficoll-Paque™ PLUS, GE Healthcare). After centrifugation at 600 × g for 15 min, the pellet containing neutrophils and red blood cells (RBCs) was subject to dextran sedimentation (3%) to separate RBCs from neutrophils. Residual RBCs were lysed by lysis buffer. Cell viability was determined by trypan blue exclusion.
9
Bovine Blood Sampling and RNA Extraction
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Peripheral blood samples (3 ml) were collected from the tail vein of cattle with the BD Vacutainer® Plus Plastic K2EDTA Tubes and BD Vacutainer® Plus Plastic Serum Tubes. The extraction of total RNA from whole blood was performed as previously described [26 (link)]. In brief, 125 μl of whole blood was mixed with the same volume of RNase-free water and 750 μl of Trizol LS reagent (Ambion) and incubated at room temperature for 5 min. Thereafter, 200 μl of chloroform (Sigma-Aldrich) was mixed and centrifuged at 13,523 ×g and 4°C for 15 min. The supernatant was collected into a 1.5 ml tube, mixed with the same volume of 70% ethanol, and then transferred to an RNAeasy column (Qiagen, Hilden, Germany) and centrifuged at 8,500 ×g for 15 sec. After the wash steps, 30 μl of RNase-free water was added and centrifuged at 8,500 ×g for 1 min. Eluted RNA was stored at -80°C until use. For the separation of serum, 3 ml of blood samples were centrifuged at 1,500 ×g for 10 min. Separated serum was transferred to 1.5 ml tube and analyzed for the presence of MAP-specific antibodies using a commercial ELISA kit.
10
Plasma Collection and Storage Protocol
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Venous blood samples were collected from each participant with a Vacutainer (BD Vacutainer Eclipse Blood Collection Needle, BD Diagnostics, Becton, Dickinson, and Company, New Jersey, USA) in 10 mL K2EDTA tubes (BD Vacutainer Plus Plastic K2EDTA Tubes, BD Diagnostics). The plasma was retrieved by centrifuging the blood samples for 30 min 1,500×g at RT immediately after collection. The plasma fraction was removed to a new tube before aliquoted and stored in − 86 °C until analysis.
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