Luminex 200 analyzer

Manufactured by Merck Group

Sourced in United States

The Luminex 200 analyzer is a multiplex assay platform that uses flow cytometry technology to detect and quantify multiple analytes simultaneously in a single sample. The instrument utilizes color-coded magnetic microspheres coated with analyte-specific capture molecules to enable the measurement of various biomolecules, such as proteins, nucleic acids, and other analytes. The Luminex 200 provides a high-throughput, reliable, and efficient method for conducting a wide range of assays, including immunoassays, protein and gene expression profiling, and molecular diagnostics.

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Luminex 200 (61 citations) Luminex analyzer (4 citations) Luminex® 200 analyzer system (4 citations) Analyzer 3.1 Luminex 200 machine (3 citations) Luminex 200 microbead analyzer (2 citations)

9 protocols using luminex 200 analyzer

Body Composition and Metabolic Markers in Clinical Trial

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Whole body dual energy X-ray absorptiometry (DXA) scans using either Hologic (Hologic Inc., Bedford, Massachusetts, USA) or GE Lunar (GE Healthcare Lunar, Madison, Wisconsin, USA) devices were performed at baseline, weeks 48 and 96. Total limb fat (upper plus lower extremities fat), trunk fat, total body lean mass and total body fat mass were quantified using standardized scanning protocols and locally calibrated based on each manufacturer’s specifications. Technicians -blinded to study clinical information- were instructed to use the same device on the same patient during the trial without central reading.
Adipokines and markers of inflammation were performed in the department of Laboratory Medicine at Hospital Universitario La Paz, Madrid on batched serum samples stored at -80°C from weeks 0 and 48. The following markers were analysed: IL-6; IL-1β; TNF-α, Insulin, Leptin, Adiponectin and Fibroblast Growth Factor-23 (FGF-23) with a multiplex magnetic bead panel assay (HBNMAG-51 Merck Millipore) using a Luminex 200 analyzer.

Bernardino J.I., Mocroft A., Wallet C., de Wit S., Katlama C., Reiss P., Mallon P.W., Richert L., Molina J.M., Knobel H., Morlat P., Babiker A., Pozniac A., Raffi F, & Arribas J.R. (2019). Body composition and adipokines changes after initial treatment with darunavir-ritonavir plus either raltegravir or tenofovir disoproxil fumarate-emtricitabine: A substudy of the NEAT001/ANRS143 randomised trial. PLoS ONE, 14(1), e0209911.

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Multiplex Analysis of Cytokine/Chemokine Profile

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Multiplex analysis based on the xMAP Luminex technology was performed with the use of MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel—Premixed 38 Plex-Immunology Multiplex Assay (sCD40L, EGF, Eotaxin/CCL11, FGF-2, Flt-3 ligand, Fractalkine, G-CSF, GM-CSF, GRO, IFN-α2, IFN-γ, IL-1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1, MCP-3, MDC/CCL22, MIP-1α, MIP-1β, TGF-α, TNF-α, TNF-β, VEGF) (Merckmillipore, Burlington, MA, USA), in accordance with the manufacturer’s instructions. Briefly, samples were incubated with fluorescent beads for 1 h, washed and incubated with phycoerythrin-streptavidin for 10 min (Merckmillipore, Burlington, MA, USA). The analysis was done using a Luminex 200 analyzer (Merckmillipore, Burlington, MA, USA). The CIMVs-MSCs and MSCs lysates in IP buffer (50 mMTris-Cl, 150 mMNaCl, 1% Nonidet-P40) were used for multiplex analysis. Equal protein load (25 μg) was used for the analysis.

Gomzikova M.O., Zhuravleva M.N., Vorobev V.V., Salafutdinov I.I., Laikov A.V., Kletukhina S.K., Martynova E.V., Tazetdinova L.G., Ntekim A.I., Khaiboullina S.F, & Rizvanov A.A. (2019). Angiogenic Activity of Cytochalasin B-Induced Membrane Vesicles of Human Mesenchymal Stem Cells. Cells, 9(1), 95.

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Biomarker Monitoring in Illness

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A full blood count was performed daily, while biochemistry tests were only performed at enrollment and then subsequently if clinically indicated. A research sample of EDTA plasma was stored every other day. All research samples were processed at the different sites within 1 h of collection, centrifuged at 500g/min for 10 min, and then stored at − 20 °C. All the samples were transferred on dry ice to OUCRU laboratory. CRP was measured on these stored samples at two time points: enrollment sample (illness day 1–3) and follow-up (day 10–21 post-symptom onset) using the same commercial assay according to the manufacturer’s specifications (magnetic bead panel, cat. no. HCVD3MAG-67 K, Merck, Millipore, UK) on a Luminex 200 analyzer.

Vuong N.L., Le Duyen H.T., Lam P.K., Tam D.T., Vinh Chau N.V., Van Kinh N., Chanpheaktra N., Lum L.C., Pleités E., Jones N.K., Simmons C.P., Rosenberger K., Jaenisch T., Halleux C., Olliaro P.L., Wills B, & Yacoub S. (2020). C-reactive protein as a potential biomarker for disease progression in dengue: a multi-country observational study. BMC Medicine, 18, 35.

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Comprehensive Plasma Biomarker Profiling

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Mice were fasted overnight before sacrifice. Blood was collected in tubes coated with EDTA followed by centrifugation at 5,000 rpm for 10 min. Plasma was collected, aliquoted and stored at −80 °C until use. The plasma concentration of the following biomarkers were quantified using Milliplex™ MAP kits and Luminex 200 Analyzer (Merck Millipore) following the manufacturer’s protocol: ghrelin with Milliplex®MAP Mouse Gut hormone Panel; sE-selectin, MMP-9, sICAM-1 and sVCAM-1 with Milliplex®MAP Mouse Cardiovascular Disease Panel I kit; IL-33 with Milliplex®MAP Mouse Cytokine/Chemokine Panel III Immunoassay respectively; with data acquisition and basic data analysis using xPONENT 3.1 and advanced data analysis using Milliplex Analyst 3.5. Plasma A-FABP and cholesterol was quantified using Mouse Adipocyte FABP ELISA Kit (BioVendor Research and Diagnostic Products Brno) and Cholesterol Assay Kit (Cayman Chemical Company) respectively following manufacturer’s protocol. Plasma endotoxin was quantified with Lonza Limulus amebocyte lysate (LAL) QCL-1000 (Lonza) following manufacturer’s protocol with heat inactivation at 75 °C for 10 min and addition of β-glucan blocker (Lonza) (1:1) to inhibit false positive readings generated by contamination from β-1,3-glucans.

Chan Y.K., Brar M.S., Kirjavainen P.V., Chen Y., Peng J., Li D., Leung F.C, & El-Nezami H. (2016). High fat diet induced atherosclerosis is accompanied with low colonic bacterial diversity and altered abundances that correlates with plaque size, plasma A-FABP and cholesterol: a pilot study of high fat diet and its intervention with Lactobacillus rhamnosus GG (LGG) or telmisartan in ApoE−/− mice. BMC Microbiology, 16, 264.

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Multiplex Cytokine Quantification from Tissue

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Frozen samples were homogenized in cold lysis buffer (300 µL per 10 mg of tissue) composed by 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% IGEPAL, and protease and phosphatase inhibitors (cOmplete, Mini Protease Inhibitor Cocktail and PhosSTOP Phosphatase Inhibitor Cocktail, Roche). Tissues were sonicated in a bath sonicator at 4 °C for 3 × 5 min. After centrifugation (12,000× g, 4 °C, 10 min), supernatants were aliquoted and stored at −80 °C until multiplex assay was performed. Total protein concentration was quantified by the Bradford method using bovine serum albumin (BSA) as a standard.
IL-1β, IL-6, IL-10, and M-CSF were simultaneously quantified in each sample using Milliplex MAP Mouse Cytokine/Chemokine Magnetic Bead Panel (Merck Millipore, Burlington, MA, USA) on a Luminex 200 analyzer (Millipore), according to the manufacturer’s protocol. Before the assay, the protein concentration of each sample was adjusted to 2 mg/mL with lysis buffer. This immunoassay uses dual-laser flow cytometry-based technology, and involves incubating the protein extract with fluorescent-coded magnetic beads pre-coated with capture antibodies, followed by sequential incubation with biotinylated detection antibodies and streptavidin-phycoerythrin conjugate [49 (link)]. Results are presented as picograms-per-mg of total protein.

Teixeira-Santos L., Veríssimo E., Martins S., Sousa T., Albino-Teixeira A, & Pinho D. (2023). Effects of NADPH Oxidase Isoform-2 (NOX2) Inhibition on Behavioral Responses and Neuroinflammation in a Mouse Model of Neuropathic Pain. Biomedicines, 11(2), 416.

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Multiplex Biomarker Immunoassay Analysis

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Serum concentrations of insulin, C-peptide, leptin, ghrelin, IL-6, and TNFα were measured using Multiplex Biomarker Immunoassays for Luminex xMAP technology (Millipore, Billerica, MA, USA; panel MMHMAG-44 k), with reading by Luminex 200 analyzer, as described [13 (link)]. These measurements were made using cardiac blood from sacrifice. All mice were fasted for 4 h prior to sacrifice.

Mahana D., Trent C.M., Kurtz Z.D., Bokulich N.A., Battaglia T., Chung J., Müller C.L., Li H., Bonneau R.A, & Blaser M.J. (2016). Antibiotic perturbation of the murine gut microbiome enhances the adiposity, insulin resistance, and liver disease associated with high-fat diet. Genome Medicine, 8, 48.

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Biomarker Profiling of Mouse Gut Hormones

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A protease inhibitor solution contained 0.1 mL Protease Inhibitor Cocktail (AEBSF, aprotinin, bestatin, EDTA, E-64, leupeptin; Sigma Aldrich), 0.1g 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF; Sigma Aldrich), and 0.9 mL DDPIV Inhibitor solution (EMD Millipore Corporation, Billerica MA). Blood collected at sacrifice was mixed 100:1 with the stock protease inhibitor. Blood cells and serum were separated by centrifugation (1000 g for 10 minutes) and serum was frozen at -70°C. Serum specimens (100 μL) were examined using a Mouse Gut Hormone Panel (EMD Millipore Corp.) using a Luminex 200 analyzer (Millipore). Two-sided Wilcoxon Rank-Sum tests were used for group comparisons.

Nobel Y.R., Cox L.M., Kirigin F.F., Bokulich N.A., Yamanishi S., Teitler I., Chung J., Sohn J., Barber C.M., Goldfarb D.S., Raju K., Abubucker S., Zhou Y., Ruiz V.E., Li H., Mitreva M., Alekseyenko A.V., Weinstock G.M., Sodergren E, & Blaser M.J. (2015). Metabolic and metagenomic outcomes from early-life pulsed antibiotic treatment. Nature Communications, 6, 7486.

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Cytokine Profiling in Murine BALF

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Lungs from control anti-IgG1-NPs, OVA anti-IgG1-NPs and OVA anti-IL4Rα-NPs groups of mice (n=6) were cannulated through the trachea and lavaged three times with 1 ml of phosphate-buffered saline containing 0.5 mM EDTA. The levels of secreted cytokines in BALF samples were then quantified using a Cytokine multiplex assay (LMC0006 mouse 20-plex panel, Novex, Invitrogen Corporation, CA, USA) and read on Luminex-200 analyzer (Millipore, Germany). Assessments of the standards, internal controls and all samples were performed in duplicate.

Halwani R., Sultana Shaik A., Ratemi E., Afzal S., Kenana R., Al-Muhsen S, & Al Faraj A. (2016). A novel anti-IL4Rα nanoparticle efficiently controls lung inflammation during asthma. Experimental & Molecular Medicine, 48(10), e262-.

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Biomarker Profiling for Kidney Toxicity

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sKIM-1 level was measured by Human Kidney Toxicity Magnetic Bead Panel 1, HKTX1MAG-38K (Millipore, Billerica, MA, USA), interleukin (IL)-6 and tumor necrosis factor alpha (TNFα) were measured by HADCYMAG-61 (Millipore, Billerica, MA, USA) on a Luminex 200 analyzer (Austin, TX, USA). FGF-21 was measured by ELISA kits (#EZHFGF21-19K; Millipore, Billerica, MA, USA) on an Infinite®M200 analyzer (Tecan Trading AG, Switzerland). The tests were performed in accordance with the manufacturer's instructions. Serum creatinine, cystatin C, glucose, triglycerides (TG), high density lipoprotein cholesterol (HDLC) and low density lipoprotein cholesterol (LDLC) tests were performed in the clinical laboratory ‘E.Gulbja laboratorija’, Riga, Latvia. The estimated glomerular filtration rate (eGFR_{EPI cyst&crea}) was determined using cystatin C and creatinine concentrations and Chronic Kidney Disease Epidemiology equation (Inker et al., 2012 (link)).
Venous blood samples for blood tests were collected without anticoagulant and were allowed to coagulate for 25–30 min at the room temperature. After that, samples were centrifuged at 4 °C for 10 min at 1600 × g. All specimens were immediately aliquoted, frozen, and stored at − 80 °C until use (for serum sKIM-1 and FGF-21 analysis; standard blood tests were performed using fresh serum samples). Serum samples were obtained after an overnight fast.

Krievina G., Tretjakovs P., Skuja I., Silina V., Keisa L., Krievina D, & Bahs G. (2016). Ectopic Adipose Tissue Storage in the Left and the Right Renal Sinus is Asymmetric and Associated With Serum Kidney Injury Molecule-1 and Fibroblast Growth Factor-21 Levels Increase. EBioMedicine, 13, 274-283.

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