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Aria 3 fusion cell sorter

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The BD ARIA III Fusion Cell Sorter is a high-performance flow cytometry instrument designed for advanced cell sorting applications. It features a multi-laser configuration and a range of detectors to enable precise sorting of complex cell populations.

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Lab products found in correlation

Fetal calf serum (fcs), by Merck Group (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Lymphoprep, by STEMCELL (2 mentions) Efluor 506, by Thermo Fisher Scientific (1 mentions) Trizol ls reagent, by Thermo Fisher Scientific (1 mentions) Sa pe, by BioLegend (1 mentions) Ez link micro sulfo nhs lc biotinylation kit, by Thermo Fisher Scientific (1 mentions) Apc sa, by BioLegend (1 mentions) Sa bv421, by BioLegend (1 mentions) Lsrfortessa flow cytometer, by BD (1 mentions)

4 protocols using aria 3 fusion cell sorter

1

Flow Cytometry-based Isolation of T-Cell Subsets

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PBMC were isolated from heparinised venous blood or buffy packs by using lymphoprep© (Stem Cell Technologies) density gradients. Resulting PBMC were frozen in liquid nitrogen and thawed to be used for subsequent experiments. Cells were sorted on an ARIA III Fusion Cell Sorter (BD Bioscience) at the University of Birmingham or FlowCore, Monash University. PBMC and purified T-cells were cultured in RPMI-1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 1% sodium pyruvate, 50 μg/mL penicillin/streptomycin (Invitrogen) and 10% fetal calf serum (Sigma). In total, Vδ1+ Teffector (CD27lo/neg) populations were sorted from 6 healthy donors, and Vδ1+ Tnaïve (CD27hi) from 3, CD8+ Tnaïve from 7, and CD8+ TEMRA populations were obtained from 9, with comparator Vδ2+ populations obtained from 2 individuals (Davey et al., 2017 (link)).
McMurray J.L., von Borstel A., Taher T.E., Syrimi E., Taylor G.S., Sharif M., Rossjohn J., Remmerswaal E.B., Bemelman F.J., Braga F.A., Chen X., Teichmann S.A., Mohammed F., Berry A.A., Lyke K.E., Williamson K.C., Stubbington M.J., Davey M.S., Willcox C.R, & Willcox B.E. (2022). Transcriptional profiling of human Vδ1 T cells reveals a pathogen-driven adaptive differentiation program. Cell reports, 39(8), 110858.
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2

Flow Cytometry-based Isolation of T-Cell Subsets

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMC were isolated from heparinised venous blood or buffy packs by using lymphoprep© (Stem Cell Technologies) density gradients. Resulting PBMC were frozen in liquid nitrogen and thawed to be used for subsequent experiments. Cells were sorted on an ARIA III Fusion Cell Sorter (BD Bioscience) at the University of Birmingham or FlowCore, Monash University. PBMC and purified T-cells were cultured in RPMI-1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 1% sodium pyruvate, 50 μg/mL penicillin/streptomycin (Invitrogen) and 10% fetal calf serum (Sigma). In total, Vδ1+ Teffector (CD27lo/neg) populations were sorted from 6 healthy donors, and Vδ1+ Tnaïve (CD27hi) from 3, CD8+ Tnaïve from 7, and CD8+ TEMRA populations were obtained from 9, with comparator Vδ2+ populations obtained from 2 individuals (Davey et al., 2017 (link)).
McMurray J.L., von Borstel A., Taher T.E., Syrimi E., Taylor G.S., Sharif M., Rossjohn J., Remmerswaal E.B., Bemelman F.J., Braga F.A., Chen X., Teichmann S.A., Mohammed F., Berry A.A., Lyke K.E., Williamson K.C., Stubbington M.J., Davey M.S., Willcox C.R, & Willcox B.E. (2022). Transcriptional profiling of human Vδ1 T cells reveals a pathogen-driven adaptive differentiation program. Cell reports, 39(8), 110858.
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3

Memory CD4+ T Cell Sorting and RNA Extraction

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Frozen PBMC were thawed and stained with fixable viability dye eFluor506 (eBiosciences) and various combinations of the antibodies listed in Supplementary file 1 as described in the flow cytometry section above. Memory CD4 T cell sorting (see gating strategy Figure 1—figure supplement 1A) was performed on a BD Aria III/Fusion cell sorter (BD Biosciences). 100,000 memory CD4+ T cells were sorted into TRIzol LS reagent (Invitrogen) for RNA extraction.
Burel J.G., Pomaznoy M., Lindestam Arlehamn C.S., Weiskopf D., da Silva Antunes R., Jung Y., Babor M., Schulten V., Seumois G., Greenbaum J.A., Premawansa S., Premawansa G., Wijewickrama A., Vidanagama D., Gunasena B., Tippalagama R., deSilva A.D., Gilman R.H., Saito M., Taplitz R., Ley K., Vijayanand P., Sette A, & Peters B. (2019). Circulating T cell-monocyte complexes are markers of immune perturbations. eLife, 8, e46045.
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4

Quantifying SARS-CoV-2 Antibody Responses

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The recombinant SARS-CoV-2 prefusion stabilized S protein and RBD were biotinylated using an EZ-Link Micro Sulfo-NHS-LC Biotinylation Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Probes were generated by coupling biotinylated proteins to fluorophore-conjugated streptavidin (SA) molecules for detection by flow cytometry (SA-BV421, SA-APC, and SA-PE, BioLegend). Isolated PBMCs were stained by a panel of antibodies listed in Table S3 as well as S protein and RBD probes (S-APC, S-PE, and RBD-BV421, 100 ng each). The samples were analyzed on a BD Aria III Fusion cell sorter (weeks 6 and 30) or a BD LSRFortessa flow cytometer (all other time points). At weeks 6 and 30, memory B cells (CD3− CD11c− CD14− CD16− CD123− HLA-DR+ CD20+ IgM− IgG+) double-positive for S protein binding were single cell sorted into 96-well plates and frozen immediately on dry ice for subsequent B cell receptor (BCR) amplification. Data were analyzed using FlowJo v.10.7.1 (FlowJo).
Lenart K., Hellgren F., Ols S., Yan X., Cagigi A., Cerveira R.A., Winge I., Hanczak J., Mueller S.O., Jasny E., Schwendt K., Rauch S., Petsch B, & Loré K. (2022). A third dose of the unmodified COVID-19 mRNA vaccine CVnCoV enhances quality and quantity of immune responses. Molecular Therapy. Methods & Clinical Development, 27, 309-323.
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