Facscanto 10c cytometer

Manufactured by BD

The FACSCanto 10c cytometer is a flow cytometry instrument designed for high-throughput cell analysis. It is capable of simultaneously detecting and analyzing up to 10 different parameters of individual cells within a sample.

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Lab products found in correlation

Trucount tube, by BD (2 mentions) Anti igd pe, by BioLegend (1 mentions) Anti pd 1 pe, by BioLegend (1 mentions) Anti igm fitc, by BioLegend (1 mentions) Facsdiva, by BD (1 mentions) Lysing solution, by BD (1 mentions) Facsdiva software v8, by BD (1 mentions)

2 protocols using facscanto 10c cytometer

Comprehensive Immune Profiling by Flow Cytometry

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All flow cytometric analyses were performed using the FACSCanto 10c cytometer (BD Biosciences, Heidelberg, Germany) and BD FACSDiva Software version 8.0.1. Cellular immune status (Panel 1), T cell activation and exhaustion (Panel 2) and B cell phenotype (Panel 3) were determined using specific markers for innate leukocytes, T cells and B cells. Briefly, cell frequencies and total cell numbers in whole blood samples were analyzed on a single-cell platform using TruCount™ tubes (BD Biosciences). Cells were stained for 30 min at room temperature using anti-CD45 allophycocyanin-H7 (APC-H7) or AlexaFluor 700 (AF-700), anti-CD3 fluorescein isothiocyanate (FITC), anti-CD8 allophycocyanin (APC), anti-CD4 peridinin chlorophyll protein complex (PerCP), anti-CD19 AF-700 or Brilliant Violet (BV510), anti-CD56 phycoerythrin (PE), anti-CD14 Brilliant Violet (BV510), anti-CD45RA BV605 and anti-CD62L BV421 or BV510, anti-CD20 APC-cyanine 7 (APC-Cy7), anti-CD27 BV421, anti-CD38 APC, anti-CD24 PerCP, anti-IgD PE, anti-IgM FITC and anti-PD-1 PE monoclonal antibodies (BioLegend and BD Biosciences) either before (Panels 1 and 2) or after (Panel 3) lysis of erythrocytes using 1x Lysing Solution according to the manufacturer’s instructions (Panels 1 and 2: BD Biosciences, Panel 3: Beckman Coulter, Brea, CA, USA).

Bonifacius A., Tischer-Zimmermann S., Dragon A.C., Gussarow D., Vogel A., Krettek U., Gödecke N., Yilmaz M., Kraft A.R., Hoeper M.M., Pink I., Schmidt J.J., Li Y., Welte T., Maecker-Kolhoff B., Martens J., Berger M.M., Lobenwein C., Stankov M.V., Cornberg M., David S., Behrens G.M., Witzke O., Blasczyk R, & Eiz-Vesper B. (2021). COVID-19 immune signatures reveal stable antiviral T cell function despite declining humoral responses. Immunity, 54(2), 340-354.e6.

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Multiparametric Flow Cytometry Analysis of T-cell Phenotypes

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Whole blood samples were transferred into BD Trucount™ tubes and extracellularly stained (Tab.S1), followed by erythrocyte lysis (Lysing Solution, BD Biosciences). For analysis of IFN-γ ELISpot assay, 1 × 10⁶ isolated PBMCs were extracellularly stained (Tab.S2) and washed. Samples were acquired on a FACSCanto™ 10c cytometer (BD Biosciences), and data were analyzed with BD FACSDiva™ Software v8.0. Gating strategies for CD3⁺, CD4⁺, and CD8⁺ T cells as well as for memory T-cell phenotypes are shown in Fig.S1A + B.

Schulze Lammers F.C., Bonifacius A., Tischer-Zimmermann S., Goudeva L., Martens J., Lepenies B., von Karpowitz M., Einecke G., Beutel G., Skripuletz T., Blasczyk R., Beier R., Maecker-Kolhoff B, & Eiz-Vesper B. (2022). Antiviral T-Cell Frequencies in a Healthy Population: Reference Values for Evaluating Antiviral Immune Cell Profiles in Immunocompromised Patients. Journal of Clinical Immunology, 42(3), 546-558.

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