Fortessa 3

Manufactured by BD

Sourced in United States

The Fortessa 3 is a high-performance flow cytometer designed for advanced cell analysis. It features a compact design and multiple laser configurations to accommodate a wide range of applications. The Fortessa 3 enables researchers to perform accurate and reliable cell counting, sorting, and analysis.

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Lab products found in correlation

Celltrace violet, by Thermo Fisher Scientific (2 mentions) Anti v5 fitc, by Thermo Fisher Scientific (1 mentions) Cellstripper, by Corning (1 mentions) Ribonuclease a, by Merck Group (1 mentions) Propidium iodide, by Merck Group (1 mentions) Celltrace cfse, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Fortessa 3 flow cytometer (2 citations) 3-or 4-Laser Fortessa (2 citations) BD LSR Fortessa III (8 citations) Fortessa (1232 citations)

4 protocols using fortessa 3

Flow Cytometry Binding Assay for T cells

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For binding assays, cancer cells were detached using CellStripper (Corning) and spleen or blood was freshly harvested from a BALB/c mouse. Red blood cells were lysed from whole blood by a 13 min. incubation in red blood cell lysis buffer containing 0.155 M ammonium chloride, 0.01 M potassium hydrogen carbonate, and 0.1 mM EDTA.
Cells were washed in PBS with 2% FBS before each incubation and kept on ice throughout. First incubation was with relevant proteins with/without 0.5 mg/mL CSA (Sigma) for 30 min, followed by incubation with anti-V5 (FITC, Invitrogen) or anti-penta-HIS (Alexa Fluor 488, Qiagen) antibodies, together with anti-CD4 (GK1.5, APC/Cy7, BioLegend) and anti-CD8 (53–6.7, APC, BD Biosciences) if detecting T cells. All flow cytometry was analyzed using LSR-II, Fortessa-3, or -5 (BD Biosciences) immediately, or the next day after cell fixation in 4% paraformaldehyde.

Skeltved N., Nordmaj M.A., Berendtsen N.T., Dagil R., Stormer E.M., Al-Nakouzi N., Jiang K., Aicher A., Heeschen C., Gustavsson T., Choudhary S., Gögenur I., Christensen J.P., Theander T.G., Daugaard M., Salanti A, & Nielsen M.A. (2023). Bispecific T cell-engager targeting oncofetal chondroitin sulfate induces complete tumor regression and protective immune memory in mice. Journal of Experimental & Clinical Cancer Research : CR, 42, 106.

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Evaluating E1 Antigen Presentation

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HEK293T cells were transfected with 2.5-3.5 µg of pURVac encoding E1, which was C-terminally extended by the SIINFEKL Ovalbumin derived CD8⁺ T cell epitope and a myc-tag and optionally fused to the Ii T cell adjuvant (25 (link)–27 (link)) at its N-terminus together with a pUC57 encoding H2Kb and β-2-microglobulin (β2m). A transfection with H2Kb alone was included as a negative control, and transfection with the H2Kb plasmid and a pUC57 plasmid encoding SIINFEKL fused directly to β2m was included as a technical positive control. 48 hours post transfection, cells were stained with PE anti-H2Kb-SIINFEKL (25-D1.16, 1:160, Invitrogen, Carlsbad, USA) and presence of SIINFEKL-H2Kb presentation on cell surfaces was detected on the LSRII or Fortessa 3 (BD Biosciences, Franklin Lakes, USA) flow cytometers, as a proxy for E1 presentation. All samples were run in biological 6-plicates, and the experiment was repeated at least two times.

Neckermann P., Boilesen D.R., Willert T., Pertl C., Schrödel S., Thirion C., Asbach B., Holst P.J, & Wagner R. (2021). Design and Immunological Validation of Macaca fascicularis Papillomavirus Type 3 Based Vaccine Candidates in Outbred Mice: Basis for Future Testing of a Therapeutic Papillomavirus Vaccine in NHPs. Frontiers in Immunology, 12, 761214.

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Analyzing Cell Cycle by Flow Cytometry

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HeLa S3 cells were harvested by centrifugation (400 × g for 4 min) and washed once with 1× PBS. Subsequently cells were fixed with 70% ice-cold ethanol and incubated for at least 30 min at 4°C. Fixed cells were washed two times with 1% BSA in PBS and incubated with 100 μg/ml ribonuclease A (Sigma, R6513) and propidium iodide (20 μg/ml) (Sigma, P4864) at 37°C for 1 h. DNA content was determined by flow cytometry with Fortessa 3 lasers (BD Biosciences) and the analysis was performed with the Flowjo software.

Pladevall-Morera D., Munk S., Ingham A., Garribba L., Albers E., Liu Y., Olsen J.V, & Lopez-Contreras A.J. (2019). Proteomic characterization of chromosomal common fragile site (CFS)-associated proteins uncovers ATRX as a regulator of CFS stability. Nucleic Acids Research, 47(15), 8004-8018.

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Peptide-specific Cytotoxicity Assay

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The assay was performed similarly to what was previously described (37 (link)). Briefly, splenocytes from naïve BALB/c mice were incubated with MfPV3 E1, E2, E6 or E7 peptide pools (same as used for immune response analyses, see above) or no peptide for 30 minutes at 37°C, 5% CO₂, 2.5 µg of each peptide/mL, and subsequently stained with combinations of 0.4 or 5 µM CellTrace CFSE and 0.2 and 2.5 µM CellTraceViolet (CTV; ThermoFisher, Waltham, USA) for 10 minutes at 37°C, 5% CO₂. Pulsed and stained splenocytes were mixed at a 1:1:1:1:1 ratio, and a total of 2.5 x10⁷ cells were injected intravenously into rAd-vaccinated recipient BALB/c mice. As the assay requires adoptive transfer of syngeneic target cells, it was necessary to use inbred mice in order to have HLA-matching. 5 hours later, spleens were harvested, and target cells were identified on the Fortessa 3 (BD Biosciences, Franklin Lakes, USA) flow cytometer by CFSE/CTV staining. The percentage of killing was calculated using the following equation:

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