Dxp11 flow cytometer

Manufactured by Cytek

Sourced in United States

The DxP11 flow cytometer is a multiparameter analytical instrument designed for the detection and analysis of biological particles, including cells and microorganisms. It utilizes principles of flow cytometry to rapidly measure and characterize multiple physical and fluorescent properties of individual particles within a sample.

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Lab products found in correlation

Intracellular fixation and permeabilization kit, by Thermo Fisher Scientific (1 mentions) Cell activation cocktail, by BioLegend (1 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions) Ionomycin, by BioLegend (1 mentions) Brefeldin a, by BioLegend (1 mentions)

2 protocols using dxp11 flow cytometer

Multiparametric Flow Cytometry Analysis of Splenocytes and Aortic Tissue

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Zombie Aqua fixable viability dye was used to eliminate nonviable cells from the analysis (Biolegend, Cat No. 422102). Single cell suspensions were stained using the following antibodies: CD11c (N418), I‐Ab (AF6‐120.1), TCRβ (H57‐597), CD86 (GL‐1), Ly‐6G (1A8), CD3 (17A2), CD4 (RM4‐4), CD8 (53‐6.7), CCR6 (29‐2L17), CD44 (IM7), CD62L (MEL14), CD11b (M1/70), TCRγδ (GL3), CD25 (PC61), CD45.2 (104), and CD115 (AFS98). Unspecific binding was reduced by co‐incubation with purified 0.5 μg anti‐CD16/CD32 per sample. Permeabilization of cells for FoxP3 staining (clone: FJK‐16S) was done using a FoxP3 Staining Buffer Set (eBioscience).
Splenocytes and aorta/perivascular adipose tissue (PVAT) were activated by Phorbol 12‐myristate 13‐acetate/ionomycin/Brefeldin A (Cell activation cocktail, Biolegend) for 4 hours after which cells were stained with extracellular antibodies and Zombie viability dye. Cells were fixed and permeabilized using intracellular fixation and permeabilization kit (eBioscience) and stained with IFN‐γ (XMG1.2) and IL‐17 (TC11‐18H10.1). All samples were acquired on a DxP11 flow cytometer (Cytek). Flow cytometry data were analyzed using FlowJo V10 analysis software (Treestar, Inc).

Engelbertsen D., Depuydt M.A., Verwilligen R.A., Rattik S., Levinsohn E., Edsfeldt A., Kuperwaser F., Jarolim P, & Lichtman A.H. (2018). IL‐23R Deficiency Does Not Impact Atherosclerotic Plaque Development in Mice. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 7(8), e008257.

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Comprehensive Immune Cell Profiling

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Single cell suspensions were analyzed by flow cytometry using the following antibodies: CD3 (17A2), CD4 (RM4-4), CD8 (53–6.7), I-Ab (AF6-120.1), CD44 (IM7), CD62L (MEL14), CD86 (GL-1), CD11b (M1/70), CD11c (N418), CD25 (PC61), NK1.1 (PK136), CD27 (LG.3A10), CD90.2 (53-2.1), Ki67 (16A8), CD127 (A7R34), CD49b (DX5), KLRG1 (2F1/KLRG1) and SIRPα (P84). Zombie Aqua fixable viability dye (Cat. No. 423102, BioLegend, San Diego (CA), USA) was used to exclude all non-viable cells from the analysis. Anti-CD16/CD32 was added to antibody mix to block non-specific mAb binding. FACS buffer containing 0.5% BSA and 0.02% sodium azide in PBS without Ca²⁺ and Mg²⁺ was used for all the washing steps.
To measure intracellular cytokines, splenocytes or aorta digested cells were stimulated with cell activation cocktail containing PMA, ionomycin and BrefeldinA (BrefA) (Cat No. 423303, Biolegend) or given Brefeldin A (Biolegend) alone for 4 hours at 37 °C. Cells were fixed and permeabilized using a fixation and permeabilization kit (Cat. No. 88-8824-00, eBioscience) and subsequently stained with IFN-γ (XMG1.2) or Granzyme B (QA16A02). FACS analysis was performed on a DxP11 flow cytometer (Cytek, Fremont (CA), USA) and the data were analyzed using FlowJo software (FlowJo LLC, Ashland (OR), USA).

Engelbertsen D., Autio A., Verwilligen R.A., Depuydt M.A., Newton G., Rattik S., Levinsohn E., Saggu G., Jarolim P., Wang H., Velazquez F., Lichtman A.H, & Luscinskas F.W. (2019). Increased lymphocyte activation and atherosclerosis in CD47-deficient mice. Scientific Reports, 9, 10608.

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