Accucount

Manufactured by Spherotech

Sourced in France

The Accucount is a laboratory instrument designed for the accurate counting and enumeration of particles or cells in a sample. It utilizes advanced optical sensing technology to provide precise and reliable particle counts.

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Lab products found in correlation

Diva software version 8, by BD (1 mentions) Fitc conjugated annexin 5, by Beckman Coulter (1 mentions) Megamix plus fsc beads, by Biocytex (1 mentions) Gallios flow cytometer, by Beckman Coulter (1 mentions) Megamix plus fsc, by Biocytex (1 mentions)

3 protocols using accucount

Comprehensive Leukocyte Immunophenotyping

Cited 1 time

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Using reverse pipetting, 100 μl of whole blood collected on EDTA-coated tubes were incubated with anti-CD32 to prevent FC-mediated non-specific binding according to the manufacturer’s instructions. The panel of antibodies as well as the gating strategy were inspired by Barnett-Vanes and al. [20 (link)] and detailed in the S1 Table. Briefly, a panel of 11 antibodies was used to identify the neutrophils (CD45+/ SSChi / His48+), the monocytes (CD45+/ SSClo/ His48hi or lo/CD43hi or lo), the B lymphocytes (CD45+/ SSClo /CD45R-B220+), the T lymphocytes (CD45+/ SSClo / CD3+/CD4+ or CD8+), and the natural killer cells (CD45+/SSClo /CD161a+). The whole blood samples were incubated with the antibodies for 20 minutes at room temperature and the BD Pharm Lyse solution was then added for 10 minutes to lyse the erythrocytes. Samples were stored at 4°C until analysis by flow cytometry (LSRII BD Biosciences). Accucount (Spherotech) were added to establish a cell count per μl of blood. Data were analyzed on Diva software version 8.0.1 (BD Biosciences).

Dupuis J., Sirois M.G., Rhéaume E., Nguyen Q.T., Clavet-Lanthier M.É., Brand G., Mihalache-Avram T., Théberge-Julien G., Charpentier D., Rhainds D., Neagoe P.E, & Tardif J.C. (2020). Colchicine reduces lung injury in experimental acute respiratory distress syndrome. PLoS ONE, 15(12), e0242318.

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Extracellular Vesicle Characterization in Cardiology

Cited 1 time

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lEVs were analyzed on a Gallios flow cytometer (Beckman Coulter)^{12 (link),14 (link)} using Megamix-Plus FSC beads (Biocytex) to define events with a 0.3- to 1-μm diameter. Phosphatidylserine externalization was assessed using FITC-conjugated annexin V (Beckman Coulter), with and without CaCl₂ (5 mmol/L). Vesicle concentration was calculated using calibrator beads (AccuCount; Spherotech).
lEVs cellular origin was identified as cardiomyocyte (caveolin 3⁺^{8 (link)}), endothelial cell (CD31⁺CD41⁻^{12 (link),14 (link)}), fibroblast (CD90.2⁺^{15 (link)}), platelet (CD41⁺^{12 (link),14 (link)}), erythrocyte (TER119⁺^{12 (link)} [glycophorin A–associated antigen]), or leukocyte (CD45⁺^{12 (link)}). Concentrations of each EV subpopulation were given per milligram of tissue. All caveolin 3+ EVs were also annexin V+.

Loyer X., Zlatanova I., Devue C., Yin M., Howangyin K.Y., Klaihmon P., Guerin C.L., Kheloufi M., Vilar J., Zannis K., Fleischmann B.K., Hwang D.W., Park J., Lee H., Menasché P., Silvestre J.S, & Boulanger C.M. (2018). Intra-Cardiac Release of Extracellular Vesicles Shapes Inflammation Following Myocardial Infarction. Circulation Research, 123(1), 100-106.

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Platelet Characterization Protocol

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(Phe-Pro-Arg chloromethl ketone) PPACK was from Haematologic Technologies Inc. (Cryopep, Montpelier, France). Anti-CD41-PE (clone P2), anti-CD235a-PE (clone 11E4B-7-6 (KC16)) monoclonal antibodies (mAbs) were from Beckman Coulter (Villepinte, France). A 1 mm SPHERO TM Ultra Rainbow and 2 mm Accu Count beads were from Spherotech (Interchim, Montluc¸on, France). Size calibrated fluorescent beads Megamix-Plus FSC were from Biocytex (Diagnostica Stago, Asnie `res-sur-Seine, France). Fluorescence calibration beads Quantum TM Cy5 MESF and Quantum TM Simply Cellular beads were from Bangs Laboratories (Polysciences Europe GmbH, Eppelheim, Germany). Anx5-Cy5 was prepared as previously described (38) . All other chemicals were of ultrapure grade (Sigma-Aldrich, Lyon, France).
A 0.22-mm filtered HEPES buffer saline (HBS) containing 150 mM NaCl, 2 mM NaN 3 , 10 mM PPACK, 0.1% BSA, 10 mM HEPES, pH 7.4 was used as dilution buffer.

Arraud N., Gounou C., Turpin D, & Brisson A.R. (2016). Fluorescence triggering: A general strategy for enumerating and phenotyping extracellular vesicles by flow cytometry. Cytometry. Part A : the journal of the International Society for Analytical Cytology, 89(2).

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