Facs aria 3

Manufactured by BioLegend

The FACS Aria III is a high-performance cell sorter that enables highly efficient cell separation and isolation. It features a multi-laser design and sophisticated optics to provide advanced analytical capabilities. The core function of the FACS Aria III is to sort and isolate specific cell populations from complex samples with high purity and recovery rates.

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Lab products found in correlation

Cd31 alexa 647, by BD (2 mentions) Ghost uv450, by Cytek Biosciences (2 mentions) Podoplanin pe, by Thermo Fisher Scientific (2 mentions) Cd45 apc cy7, by BioLegend (2 mentions) Legendplex immunoassay, by BioLegend (1 mentions)

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FACS Aria

3 protocols using facs aria 3

Multiplex Cytokine Profiling of MoDC

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MoDC culture supernatants were collected after chemical treatment for 24 h. Cells and debris were removed by centrifugation for 10 min at 300 g. Following the manufacturer's instructions, IL-1β, IL-6, IL-8, IL-10, IL12p70, IL-18 and TNFα were quantified with a customized bead-based LEGENDplex immunoassay (Biolegend) on a FACS Aria III. Data were analyzed using the provided LEGENDplex v8.0 software. Data were log2 transformed and visualized with GraphPad prism. Student's t-test was applied to calculate p-values relative to medium control.

Höper T., Siewert K., Dumit V.I., von Bergen M., Schubert K, & Haase A. (2021). The Contact Allergen NiSO4 Triggers a Distinct Molecular Response in Primary Human Dendritic Cells Compared to Bacterial LPS. Frontiers in Immunology, 12, 644700.

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Isolation and Characterization of Lymphatic Endothelial Cells

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After generating a single cell suspension and staining for cpLECs with Ghost UV450 (Tonbo Biosciences, Catalog #: 13-0868-T500), CD31 Alexa 647 (BD Biosciences, Catalog #: 565608), Podoplanin PE (eBioscience, Catalog #: 12-5381-82), and CD45 APC-Cy7 (BioLegend, Catalog #: 103116), the cells were sorted with a FACS Aria III with a nozzle size of 130 μm at the UW Flow Core satellite facility in the UW Biotechnology Center. LECs were gated for singlets using both FSC and SSC, dead cells excluded by gating for Ghost^negative live cells, and LECs were identified using both CD31⁺ Podoplanin⁺ after excluding both CD45^intermediate microglia and CD45⁺ leukocytes as previously described^{2 (link),6 (link),8 (link),19}. Cells were sorted into either FACS buffer (1% BSA in PBS) for scRNA-seq, or RPMI containing 10% FBS, 2-mercaptoethanol, and penicillin/streptomycin for in vitro co-culture experiments.

Hsu M., Laaker C., Madrid A., Herbath M., Choi Y.H., Sandor M, & Fabry Z. (2022). Neuroinflammation creates an immune regulatory niche at the meningeal lymphatic vasculature near the cribriform plate. Nature immunology, 23(4), 581-593.

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Single-cell RNA-seq of cpLECs

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After generating a single cell suspension and staining for cpLECs with Ghost UV450 (Tonbo Biosciences, Catalog #: 13-0868-T500), CD31 Alexa 647 (BD Biosciences, Catalog #: 565608), Podoplanin PE (eBioscience, Catalog #: 12-5381-82), and CD45 APC-Cy7 (Biolegend, Catalog #: 103116) as previously described, the cells were sorted with a FACS Aria III with a nozzle size of 130 um at the UW Flow Core satellite facility in the UW Biotechnology Center. Cells were sorted into FACS buffer (1% BSA in PBS). A total of 25,102 cells from the control group and 53,224 cells from the experimental group post-sort were provided to the Biotechnology Core for scRNAseq.

Hsu M., Madrid A., Choi Y.H., Laaker C., Herbáth M., Sándor M., & Fábry Z. (2020). Neuroinflammation alters the phenotype of lymphangiogenic vessels near the cribriform plate.

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