Facs aria 3 high speed cell sorter

Manufactured by BD

Sourced in United States

The BD FACS Aria III is a high-speed cell sorter that enables the isolation and collection of specific cell populations from complex samples. It utilizes flow cytometry technology to rapidly analyze and sort individual cells based on their physical and fluorescent characteristics.

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Lab products found in correlation

Trypsin, by Merck Group (2 mentions) Facscanto 2 cytometer, by BD (2 mentions) Hank s balanced salt solution, by Merck Group (1 mentions) Heat inactivated human serum, by Merck Group (1 mentions) Cts optmizer t cell expansion sfm, by Thermo Fisher Scientific (1 mentions) Fc block, by Thermo Fisher Scientific (1 mentions) Cd4 pe cy7, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by BD (1 mentions) Cell staining buffer, by BioLegend (1 mentions) Human trustain fcx, by BioLegend (1 mentions) Annexin 5 fitc double staining apoptosis detection kit, by Beyotime (1 mentions)

Spelling variants of this product

Aria III (342 citations) Aria cell sorter (91 citations) BD Aria sorter (37 citations) Aria III sorter (32 citations) FACS Aria High-Speed Cell Sorter (14 citations) FACS Aria cell (2 citations) BD FACS III (2 citations) BD FACSTM (2 citations)

16 protocols using facs aria 3 high speed cell sorter

Detecting BVDV Infection and Apoptosis

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Flow cytometry analysis was performed to detect the intracellular BVDV protein, cells were incubated with anti-BVDV-NS4B antibody and stained with FITC-conjugated anti-mouse IgG antibody. Then, the cells were analyzed using BD FACS Aria™ III High Speed Cell Sorter (BD Biosciences, San Diego, CA, USA).
Apoptosis analysis was performed using the annexin V-FITC double-staining apoptosis detection kit (Beyotime, C1062S) according to the manufacturer’s instructions. Briefly, MDBK cells were infected with NCP BVDV in the presence or absence of siParkin or siNC, the cells were resuspended in 500 μL of 10× binding buffer, then stained with 5 μL of FITC-labelled annexin V and 10 μL of propidium iodide (PI) for 15 min. Subsequently, fluorescence intensity of all staining samples was measured using BD FACS Aria™ III High Speed Cell Sorter (BD Biosciences, San Diego, CA, USA), followed by analysis with FlowJo software, version 10 (Treestar, San Carlos, CA, USA).

Li Z., Zhang Y., Zhao B., Xue Q., Wang C., Wan S., Wang J., Chen X, & Qi X. (2024). Non-cytopathic bovine viral diarrhea virus (BVDV) inhibits innate immune responses via induction of mitophagy. Veterinary Research, 55, 27.

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High-Speed FACS Sorting of GFP+ MEFs

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Cell sorting was carried out using a BD FACSAria III high speed cell sorter. GFP positive MEFs were analyzed at a velocity of 2000 cells/s, and 2000 cells were sorted into a culture plate well with DMEM.

Berthelsen M.F., Leknes S.L., Riedel M., Pedersen M.A., Joseph J.V., Hager H., Vendelbo M.H, & Thomsen M.K. (2021). Comparative Analysis of Stk11/Lkb1 versus Pten Deficiency in Lung Adenocarcinoma Induced by CRISPR/Cas9. Cancers, 13(5), 974.

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RPE Flat-Mounting and FACS Analysis

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Mice were euthanized by cervical dislocation. For histological analysis of wholemounts, eyes were prepared as previously described.⁴⁴ (link) In brief, eyes were enucleated, cleaned, and fixed in fresh 4% paraformaldehyde for 2 h at RT. The cornea, lens, and neuroretina were removed. Eight incisions from the periphery to the optic nerve enabled flat mounting of the tissue with the RPE cells facing upward on a glass slide. For FACS analysis, the enucleated eyes were cleansed and placed in 1% trypsin (Sigma-Aldrich) for 1 h at 37°C. Cells were rinsed off the eyecup using a syringe and a 27-gauge needle with HBSS. The resulting single-cell solution was sorted on a FACSAria III high-speed cell sorter (BD Biosciences, San Jose, CA, USA) using the GFP detector. Cells were initially separated using FSC and SSC as indicated in Figure S5. FSC and SSC doublets were excluded prior to sorting of cells according to detected GFP levels. No cell-specific marker proteins were utilized.

Holmgaard A.B., Askou A.L., Jensen E.G., Alsing S., Bak R.O., Mikkelsen J.G, & Corydon T.J. (2020). Targeted Knockout of the Vegfa Gene in the Retina by Subretinal Injection of RNP Complexes Containing Cas9 Protein and Modified sgRNAs. Molecular Therapy, 29(1), 191-207.

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Isolation of eGFP+ Retinal Cells

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Mice were euthanized, and the eyes were enucleated and dissected immediately. The lens, cornea, and neuroretina were removed, and the eyecup was placed in 1% trypsin (Sigma-Aldrich) for 1 hr at 37°C. Cells in the eyecup were rinsed in Hank’s balanced salt solution (Sigma-Aldrich), and the cell population was placed in 1% trypsin for 1 hr to obtain a single-cell solution. The solution was sorted on a FACSAria III high-speed cell sorter (BD Biosciences, San Jose, CA, USA) for sorting of eGFP⁺ cells. During the sorting, no cells were excluded due to aggregation, size, or debris. The only sorting criterion was eGFP expression.

Holmgaard A., Askou A.L., Benckendorff J.N., Thomsen E.A., Cai Y., Bek T., Mikkelsen J.G, & Corydon T.J. (2017). In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells. Molecular Therapy. Nucleic Acids, 9, 89-99.

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GFP Expression Analysis by Flow Cytometry

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GFP expression in cells transfected with the various reporter plasmids was analysed by flow cytometry using a FACS Canto II cytometer (BD Biosciences). Briefly, viable cells were first gated based on their forward (FSC) and side scatter (SCC) properties. Cell doublets were excluded using FSC‐width vs. FSC‐height. Old cells expressing high levels of GFP were sorted from old cells with low GFP fluorescence (fraction V elutriation) using a FACS Aria III high speed cell sorter (BD Biosciences) with a 70 μm nozzle. Sorting was performed at 4°C. The cut‐off for GFP‐high cells was defined based on cells transformed with an empty control plasmid.

Streubel M.K., Bischof J., Weiss R., Duschl J., Liedl W., Wimmer H., Breitenbach M., Weber M., Geltinger F., Richter K, & Rinnerthaler M. (2017). Behead and live long or the tale of cathepsin L. Yeast (Chichester, England), 35(2), 237-249.

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Isolation and Expansion of Naïve Tregs

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Naïve Tregs were isolated as described previously [12 (link)]. Human CD4⁺ T cells were stained with cyanine 5–anti-CD4, fluorescein isothiocyanate (FITC)–anti-CD25, and phycoerythrin (PE)–anti-CD45RA. The stained Tregs were sorted on the basis of the CD4⁺CD25⁺CD45RA⁺ T-cell phenotypic markers using a fluorescence-activated cell sorting (FACS) machine (FACSAria III high-speed cell sorter, BD Biosciences, San Jose, CA, USA) and were incubated in OpTmizer CTS T-cell Expansion SFM (Thermo Fisher Scientific Inc., Rockford, IL, USA) containing 2% heat-inactivated human serum (Sigma-Aldrich, St. Louis, MO, USA).

Joo J.Y., Cha G.S., Kim H.J., Lee J.Y, & Choi J. (2020). Atheroprotective nasal immunization with a heat shock protein 60 peptide from Porphyromonas gingivalis. Journal of Periodontal & Implant Science, 50(3), 159-170.

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High-Speed Cell Sorting Optimization

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Cell sorting was carried out using a BD FACSAria III high-speed cell sorter. GFP or YFP positive cells were analysed at a velocity of 2000 cells/s and sorted into a culture plate with DMEM ranging from 1 to 2000 cells per well at 4 °C.

Berthelsen M.F., Riedel M., Cai H., Skaarup S.H., Alstrup A.K., Dagnæs-Hansen F., Luo Y., Jensen U.B., Hager H., Liu Y., Callesen H., Vendelbo M.H., Jakobsen J.E, & Thomsen M.K. (2021). The CRISPR/Cas9 Minipig—A Transgenic Minipig to Produce Specific Mutations in Designated Tissues. Cancers, 13(12), 3024.

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Flow Cytometry-based GFP Sorting

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GFP expression in cells transfected with the various reporter plasmids was analysed by flow cytometry using a FACS Canto II cytometer (BD Biosciences). Briefly, viable cells were first gated based on their forward (FSC) and side scatter (SCC) properties. Cell doublets were excluded using FSC-width vs. FSC-height. Old cells expressing high levels of GFP were sorted from old cells with low GFP fluorescence (fraction V elutriation) using a FACS Aria III high speed cell sorter (BD Biosciences) with a 70 μm nozzle. Sorting was performed at 4°C. The cut-off for GFP-high cells was defined based on cells transformed with an empty control plasmid.

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Plasma Microvesicle Profiling by Flow Cytometry

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Flow cytometric analysis of plasma MV content was performed according to a recently described protocol [19 (link)] on a BD FACSAria™ III High Speed Cell Sorter (BD Biosciences, San Jose, CA, USA) at a maximal rate of 2 × 10⁴ events per second until 10⁶ events were collected in total. Data processing was performed as demonstrated in Figure 1. A size gate encompassing approximately 100 nm to 1000 nm was established in FlowJo™ software (v. 10.0.8, Tree Star, Inc., Oregon, USA) using the “Autogate” function around 200 nm and 900 nm fluorescent, size calibrated beads on log-scaled forward scatter height (FSC-H), and side scatter height (SSC-H). In addition, a discriminator was set at 200 on SSC-H to avoid exclusion of the smallest events. MVs were defined as phosphatidylserine positive (PS+) events based on binding of lactadherin-FITC within the established size gate. MVs of platelet origin were defined as CD41 positive events; monocyte MVs (MMVs) were defined as CD14 positive events; endothelial MVs (EMVs) were defined as CD62E positive events, and the expression of CD36 was investigated on PMVs, MMVs, and EMVs.

Botha J., Velling Magnussen L., Nielsen M.H., Nielsen T.B., Højlund K., Andersen M.S, & Handberg A. (2017). Microvesicles Correlated with Components of Metabolic Syndrome in Men with Type 2 Diabetes Mellitus and Lowered Testosterone Levels But Were Unaltered by Testosterone Therapy. Journal of Diabetes Research, 2017, 4257875.

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Isolation and Purification of Th1 and Th17 Cells

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Twelve to 16 weeks after HFD or NCD, splenic CD4+ T cells were isolated by magnetic cell separation (MACS) using the Miltenyi kit. The purified CD4+ T cells were incubated with Fc-block (Thermo Fisher Scientific) followed by staining with CD4-PE-Cy7, CXCR3-FITC, CCR4-PE, and CCR6-APC for 30 min in the dark. All antibodies were obtained from Biolegend (Fell, Germany). Dead cells were excluded by DAPI (BD Biosciences) staining and Th1 cells (CD4+CXCR3+CCR6–) and Th17 cells (CD4+CCR4+CCR6+CXCR3–) were purified using the BD FACS Aria III high-speed cell sorter (BD Biosciences).

Surendar J., Frohberger S.J., Karunakaran I., Schmitt V., Stamminger W., Neumann A.L., Wilhelm C., Hoerauf A, & Hübner M.P. (2019). Adiponectin Limits IFN-γ and IL-17 Producing CD4 T Cells in Obesity by Restraining Cell Intrinsic Glycolysis. Frontiers in Immunology, 10, 2555.

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