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Lithium heparin vacutainer

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Lithium heparin vacutainers are blood collection tubes used to collect and store blood samples. They contain the anticoagulant lithium heparin, which prevents the blood from clotting during the collection and transportation process.

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Lab products found in correlation

Histopaque 1077, by Merck Group (4 mentions) Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (4 mentions) Opti mem, by Thermo Fisher Scientific (4 mentions) Dulbecco s phosphate buffered saline dpbs ca2 mg2 free, by Thermo Fisher Scientific (2 mentions) Odn2216 and chemically synthesized rnas, by Integrated DNA Technologies (2 mentions) Multiplex chemiluminescence plates for the detection, by Quansys Biosciences (2 mentions) Dnase 1, by New England Biolabs (2 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions) Quickextract dna extraction solution, by Illumina (2 mentions) Megaclear transcription clean up kit, by Thermo Fisher Scientific (2 mentions) Antarctic phosphatase, by New England Biolabs (2 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle medium dmem, by American Type Culture Collection (2 mentions) Kapa hifi hotstart dna polymerase, by Roche (2 mentions) Geneticin, by Thermo Fisher Scientific (2 mentions) Hiscribe t7 high yield rna synthesis kit, by New England Biolabs (2 mentions) Trypan blue, by Thermo Fisher Scientific (2 mentions) Dulbecco s phosphate buffered saline, by Merck Group (2 mentions) Sodium heparin vacutainer, by BD (1 mentions) Easysep direct human neutrophil isolation kit, by STEMCELL (1 mentions)

28 protocols using lithium heparin vacutainer

1

Isolation of Immune Cells from Blood

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Neutrophils were isolated from fresh venous blood collected in lithium heparin vacutainers (BD, Scoresby, Australia) using the EasySep Direct Human Neutrophil Isolation Kit (STEMCELL Technologies, Tullamarine, Australia) as per manufacturer’s instructions. Neutrophil purity was assessed by cell morphology using light microscopy of Giemsa stained smears of the isolated cells, and viability was assessed using trypan blue exclusion. Monocytes were isolated from both fresh venous blood, collected in lithium heparin vacutainers (BD), as well as from buffy coats supplied from the Australian Red Cross Blood Service. Monocytes were isolated by negative selection using the RosetteSep Human Monocyte Enrichment Cocktail (STEMCELL Technologies) as per manufacturer’s instructions. Monocyte purity was assessed by staining (anti-CD14 antibody, BioLegend, San Diego, CA) and measuring CD14+ cells by flow cytometry. Monocytes were either frozen in fetal bovine serum (FBS) in 20% dimethyl sulfoxide (DMSO) in liquid nitrogen for later use or used immediately after isolation. Natural killer (NK) cells were isolated from fresh venous blood collected in sodium heparin vacutainers (BD). NK cells were isolated by negative selection using the RosetteSep Human NK Enrichment Cocktail (STEMCELL Technologies) as per manufacturer’s instructions.
Aitken E.H., Damelang T., Ortega-Pajares A., Alemu A., Hasang W., Dini S., Unger H.W., Ome-Kaius M., Nielsen M.A., Salanti A., Smith J., Kent S., Hogarth P.M., Wines B.D., Simpson J.A., Chung A.W, & Rogerson S.J. (2021). Developing a multivariate prediction model of antibody features associated with protection of malaria-infected pregnant women from placental malaria. eLife, 10, e65776.
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2

Optimized Cell Assay Protocols

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Lithium heparin vacutainers were purchased from BD Biosciences (San Jose, CA). RPMI, fetal bovine serum (FBS), penicillin-streptomycin, Dulbecco’s phosphate-buffered saline (DPBS; Ca2+/Mg2+-free), Hank’s balanced salt solution (HBSS), MEGAclear™ Transcription Clean-Up Kit, Geneticin, Opti-MEM®, and Lipofectamine® RNAiMAX transfection reagent were purchased from Thermo Fisher Scientific (Waltham, MA). ODN2216 and chemically synthesized RNAs were synthesized by Integrated DNA Technologies, Inc. (IDT; Coralville, IA). A HiScribe™ T7 High Yield RNA Synthesis Kit, DNase I, and Antarctic Phosphatase were purchased from New England Biolabs (Ipswitch, MA). Multiplex chemiluminescence plates for the detection of type I IFNs were custom-manufactured by Quansys Biosciences (Logan, UT). Dulbecco’s Modified Eagle Medium (DMEM) was purchased from ATCC (Manassas, VA). QuickExtract™ DNA Extraction Solution was purchased from Epicentre (Madison, WI). KAPA HiFi HotStart DNA Polymerase was purchased from Kapa Biosystems (Wilmington, MA). A Mutation Discovery Kit for the Fragment Analyzer™ was purchased from Advanced Analytical Technologies, Inc. (Ames, IA).
Schubert M.S., Cedrone E., Neun B., Behlke M.A, & Dobrovolskaia M.A. (2018). Chemical Modification of CRISPR gRNAs Eliminate type I Interferon Responses in Human Peripheral Blood Mononuclear Cells. Journal of cytokine biology, 3(1), 121.
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3

Influenza Vaccine Immune Response Kinetics

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All donors received the trivalent inactivated influenza vaccine Mutagrip 2011/2012 (Sanofi-Pasteur) intramuscularly by a physician. The vaccine was composed of the following strains recommended by the WHO for the 2011/2012 influenza season in the northern hemisphere: an A/California/7/2009 (H1N1)-like virus, an A/Perth/16/2009 (H3N2)-like virus, a B/Brisbane/60/2008-like virus [38 ].
From each donor, a total of 50 ml of blood was drawn before injection of the vaccine and on day 3, 7, 10, 14, 17, and 21 after vaccination using Lithium-Heparin Vacutainers (BD Biosciences). The blood samples were processed immediately.
Stervbo U., Pohlmann D., Baron U., Bozzetti C., Jürchott K., Mälzer J.N., Nienen M., Olek S., Roch T., Schulz A.R., Warth S., Neumann A., Thiel A., Grützkau A, & Babel N. (2017). Age dependent differences in the kinetics of γδ T cells after influenza vaccination. PLoS ONE, 12(7), e0181161.
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4

Genomic DNA Isolation from Blood

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Blood samples (∼1 ml) were collected in lithium-heparin vacutainers (BD Biosciences, San Jose, CA, USA) after written informed consent was received from the volunteers. Genomic DNA was isolated from peripheral blood by a Qiagen DNA Blood Minikit (Qiagen, Hilden, Germany) as per the manufacturer’s protocol.
Mandal A., Kumar M., Kumar A., Sen A., Das P, & Das S. (2021). TLR4 and TLR9 polymorphism: Probable role in susceptibility among the population of Bihar for Indian visceral leishmaniasis. Innate Immunity, 27(6), 493-500.
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5

Plasma Nitrite and Nitrate Quantification

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Whole blood samples were collected via cannula into two 6-mL lithium heparin vacutainers (BD). One lithium heparin tube was centrifuged immediately at 5000 rpm (2795 × g; 4°C) for 3 min; the plasma was divided into aliquots in dark-colored Eppendorf tubes and frozen at −80°C to preserve the NO2−. These Eppendorf tubes were pretreated with a stop solution to prevent transition between compounds, as described by Nagababu and Rifkind (16 (link)). The remaining vacutainer was kept cool and processed within 30 min. The sample was then centrifuged at 3000 rpm (1006 × g) for 10 min at 4°C and the plasma was divided into aliquots and frozen at −80°C. The plasma samples were used to measure NO3− and NO2− concentrations.
Capper T.E., Siervo M., Clifford T., Taylor G., Iqbal W., West D, & Stevenson E.J. (2021). Pharmacokinetic Profile of Incremental Oral Doses of Dietary Nitrate in Young and Older Adults: A Crossover Randomized Clinical Trial. The Journal of Nutrition, 152(1), 130-139.
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6

Isolation of PBMCs from Venous Blood

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Venous blood (18 mL) was collected into lithium heparin vacutainers (BD Biosciences), diluted with Dulbecco’s phosphate-buffered saline (Sigma-Aldrich) at a ratio of 1:1, and layered on top of Histopaque-1077 (Sigma-Aldrich) in SepMate-50 (IVD) tubes (Stemcell Technologies, Canada). After centrifugation for 15 minutes at 1200g, the upper layer containing plasma and PBMCs were washed twice with Dulbecco’s phosphate-buffered saline. PBMCs were then counted using trypan blue (Gibco, United Kingdom).
Malik A., Sayed A.A., Han P., Tan M.M., Watt E., Constantinescu-Bercu A., Cocker A.T., Khoder A., Saputil R.C., Thorley E., Teklemichael A., Ding Y., Hart A.C., Zhang H., Mitchell W.A., Imami N., Crawley J.T., Salles-Crawley I.I., Bussel J.B., Zehnder J.L., Adams S., Zhang B.M, & Cooper N. (2023). The role of CD8+ T-cell clones in immune thrombocytopenia. Blood, 141(20), 2417-2429.
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7

Trivalent Influenza Vaccine Response

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The vaccination was performed intramuscularly by a study physician with the trivalent influenza vaccine (Mutagrip 2011/2012 Sanofi-Pasteur). The vaccine was composed of A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), B/Brisbane/60/2008 according to WHO recommendation. 50 ml venous blood was drawn at day 0, 7, 14, and 21 post vaccination using Lithium-Heparin Vacutainers (BD Biosciences) and processed immediately.
Nienen M., Stervbo U., Mölder F., Kaliszczyk S., Kuchenbecker L., Gayova L., Schweiger B., Jürchott K., Hecht J., Neumann A.U., Rahmann S., Westhoff T., Reinke P., Thiel A, & Babel N. (2019). The Role of Pre-existing Cross-Reactive Central Memory CD4 T-Cells in Vaccination With Previously Unseen Influenza Strains. Frontiers in Immunology, 10, 593.
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8

Cytokine Release Assay for Immune Modulators

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Blood from healthy human volunteers was collected into lithium heparin vacutainers (BD) and used within 2 h of the blood draw. Minimally-diluted blood was stimulated with either 100 μg/mL of IgG1 Fc hexamer or 10 μg/mL Campath (Genzyme). Briefly, 12.5 μL of 20x final concentration Avastin (5 μg/mL), OKT3 (5 μg/mL), TGN1412 (5 μg/mL), Campath (5 μg/mL), or IgG1 Fc Hexamer (100 μg/mL) was transferred to a 96-well round bottom tissue culture plate (Costar). 237.5 μL of whole blood was added and mixed gently by pipetting. Plates were incubated at 37°C, 5 % CO2, 100 % humidity for 24 h, centrifuged at 300 g for 5 min and plasma collected for cytokine analysis. Plasma not analyzed immediately was stored at −80°C until analysis.
Hussain K., Hargreaves C.E., Rowley T.F., Sopp J.M., Latham K.V., Bhatta P., Sherington J., Cutler R.M., Humphreys D.P., Glennie M.J., Strefford J.C, & Cragg M.S. (2019). Impact of Human FcγR Gene Polymorphisms on IgG-Triggered Cytokine Release: Critical Importance of Cell Assay Format. Frontiers in Immunology, 10, 390.
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9

Controlled Malaria Infection Study

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Inoculum preparation, volunteer recruitment, infection, monitoring and treatment were performed as described previously [64 (link)]. In brief, healthy malaria-naive individuals were intravenously inoculated with 1,800 viable P. falciparum parasitized erythrocytes, and treated with anti-malarial drugs at seven to eight days post-infection. Blood samples were collected prior to infection and seven days post-infection (before anti-malarial drug treatment). Peripheral blood from healthy volunteers was also collected under informed consent and approval by the QIMRB-HREC. Peripheral blood collected in Lithium Heparin Vacutainers (BD Biosciences) was either used directly for flow cytometry analysis, or peripheral blood mononuclear cells (PBMC) isolated using standard Ficoll density gradient centrifugation.
Burel J.G., Apte S.H., Groves P.L., Klein K., McCarthy J.S, & Doolan D.L. (2016). Reduced Plasmodium Parasite Burden Associates with CD38+ CD4+ T Cells Displaying Cytolytic Potential and Impaired IFN-γ Production. PLoS Pathogens, 12(9), e1005839.
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10

Optimized Cell Assay Protocols

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Lithium heparin vacutainers were purchased from BD Biosciences (San Jose, CA). RPMI, fetal bovine serum (FBS), penicillin-streptomycin, Dulbecco’s phosphate-buffered saline (DPBS; Ca2+/Mg2+-free), Hank’s balanced salt solution (HBSS), MEGAclear™ Transcription Clean-Up Kit, Geneticin, Opti-MEM®, and Lipofectamine® RNAiMAX transfection reagent were purchased from Thermo Fisher Scientific (Waltham, MA). ODN2216 and chemically synthesized RNAs were synthesized by Integrated DNA Technologies, Inc. (IDT; Coralville, IA). A HiScribe™ T7 High Yield RNA Synthesis Kit, DNase I, and Antarctic Phosphatase were purchased from New England Biolabs (Ipswitch, MA). Multiplex chemiluminescence plates for the detection of type I IFNs were custom-manufactured by Quansys Biosciences (Logan, UT). Dulbecco’s Modified Eagle Medium (DMEM) was purchased from ATCC (Manassas, VA). QuickExtract™ DNA Extraction Solution was purchased from Epicentre (Madison, WI). KAPA HiFi HotStart DNA Polymerase was purchased from Kapa Biosystems (Wilmington, MA). A Mutation Discovery Kit for the Fragment Analyzer™ was purchased from Advanced Analytical Technologies, Inc. (Ames, IA).
Schubert M.S., Cedrone E., Neun B., Behlke M.A, & Dobrovolskaia M.A. (2018). Chemical Modification of CRISPR gRNAs Eliminate type I Interferon Responses in Human Peripheral Blood Mononuclear Cells. Journal of cytokine biology, 3(1), 121.
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