Fortessa flow cytometer

Manufactured by Tree Star

The Fortessa flow cytometer is a versatile and powerful instrument used for the analysis and sorting of cells and other biological particles. It utilizes the principles of flow cytometry to rapidly measure and analyze multiple physical and fluorescent characteristics of individual cells or particles passing through a laser-based detection system. The Fortessa provides high-performance data acquisition and analysis capabilities, enabling researchers to gain insights into various cell populations and their properties.

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Lab products found in correlation

Mitotracker deep red, by Thermo Fisher Scientific (1 mentions) Zombie aqua, by BioLegend (1 mentions) Mitotracker green, by Thermo Fisher Scientific (1 mentions) Lsr 2, by BD (1 mentions) Foxp3 staining kit, by Thermo Fisher Scientific (1 mentions) Anti cd4 percp cy5.5 clone rm4 5, by BD (1 mentions) Anti cd44 apc cy7 clone im7, by BD (1 mentions) Ki67 pe clone b56, by BD (1 mentions)

Close spelling variants of this product

Fortessa (6 citations) LSR-II/Fortessa flow cytometer (6 citations)

6 protocols using fortessa flow cytometer

Isolation and Analysis of Murine Splenocytes

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The study was approved by and mouse handling complies with all relevant ethical regulations of the Australian National University Ethics Comittee. Spleens were isolated as single-cell suspensions after red blood cell lysis. To stain for surface markers, we incubated cells in the antibody mixture diluted in ice-cold staining buffer (2% fetal calf serum in phosphate-buffered saline). Ramos cells were loaded with Indo-1 AM at 37 °C for 2 h before being stimulated at 37° with anti-Fab(2) antibody. An LSRII or Fortessa Flow Cytometer with FACSDiva software were used for flow cytometry acquisition, and FlowJo (Tree Star) was used for analysis.

Jiang S.H., Athanasopoulos V., Ellyard J.I., Chuah A., Cappello J., Cook A., Prabhu S.B., Cardenas J., Gu J., Stanley M., Roco J.A., Papa I., Yabas M., Walters G.D., Burgio G., McKeon K., Byers J.M., Burrin C., Enders A., Miosge L.A., Canete P.F., Jelusic M., Tasic V., Lungu A.C., Alexander S.I., Kitching A.R., Fulcher D.A., Shen N., Arsov T., Gatenby P.A., Babon J.J., Mallon D.F., de Lucas Collantes C., Stone E.A., Wu P., Field M.A., Andrews T.D., Cho E., Pascual V., Cook M.C, & Vinuesa C.G. (2019). Functional rare and low frequency variants in BLK and BANK1 contribute to human lupus. Nature Communications, 10, 2201.

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Isolation and Analysis of Retinal Cells

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Cell suspensions were prepared from freshly dissected retinal tissue. SoFa1 embryos (72 hpf) were transferred to cold Ca²⁺-free medium [116.6 mM NaCl, 0.67 mM KCl, 4.62 mM Tris; 0.4 mM EDTA (pH 7.8) (Harris and Messersmith, 1992 (link))] supplemented with 100 µg/ml of heparin and 0.04% MS-222. Fifteen to 20 eyes were transferred to glass Petri dishes. Without disturbing the retinas, the Ca²⁺-free medium was removed and 0.25% Trypsin-EDTA was added. Following a 10 min incubation at 37°C, the trypsin was removed, and the retinas were mechanically dissociated by pipetting using flame-pulled glass Pasteur pipettes. For confocal imaging, single cell suspensions were plated into 35 mm imaging dishes, seeded for 1 h at 28.5°C and imaged. For flow cytometry, single cell suspensions were analysed using a Fortessa flow cytometer and analysed using FlowJo (Tree Star). Retinal cells were discriminated based on the forward (FSC-A) and side-scatter (SSC-A) profiles. Single-colour RFP, CFP and GFP cells were used to generate a compensation matrix.

Almeida A.D., Boije H., Chow R.W., He J., Tham J., Suzuki S.C, & Harris W.A. (2014). Spectrum of Fates: a new approach to the study of the developing zebrafish retina. Development (Cambridge, England), 141(9), 1971-1980.

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Intracellular Cytokine Staining of Splenocytes

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Two weeks after the final immunization or eight weeks after live genital C. muridarum immunization, mice were sacrificed and splenocytes were stimulated with 2 μg/ml antibody to CD28 and HK-EBs (5 × 10⁵ IFU/ml) in complete RPMI 1640 for 4 h at 37°C. Brefeldin A was added at a final concentration of 1 μg/ml, and cells were incubated for an additional 12 h before intracellular cytokine staining. The protocol of intracellular cytokine staining was previously described [24 (link)]. We acquired 200,000 live lymphocytes per sample by using a Fortessa flow cytometer and analyzed the data by using the FlowJo software program (Tree Star).

Yu H., Karunakaran K.P., Jiang X, & Brunham R.C. (2014). Evaluation of a multisubunit recombinant polymorphic membrane protein and major outer membrane protein T cell vaccine against Chlamydia muridarum genital infection in three strains of mice. Vaccine, 32(36), 4672-4680.

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Phenotypic Characterization of Porcine BAL Cells

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Pig BAL cells or migrated cells from pig skin biopsies were incubated for 30 min on ice in saturation buffer made of PBS supplemented with 5% horse serum and 5% swine serum. For analysis of biot-anti-CD11c binding on BAL cells, biot-anti-CD11c and biot-ISC were used at 2 µg/ml followed by Alexa 647-conjugated streptavidin. For labeling with VC, VC were first made on ice, and VC corresponding to 1.5 µg/ml biot-mAb + 6 µg/ml SA-NP were added on the cells for 30 min. The cells were then washed and successively incubated with rabbit anti-NP IgG and anti-MHC class 2 (anti-SLA-DR, MSA3 mAb, IgG2a, 2 µg/ml) and followed by highly specific Alexa 647-conjugated goat anti-rabbit IgG (10 µg/ml) and Alexa 488-conjugated goat anti-mouse IgG2a (2 µg/ml). Dead cells were excluded by DAPI labeling. Flow cytometry acquisition was done with a Becton Dickinson Fortessa Flow cytometer and the acquired data were analyzed using FlowJo software (version X.0.6, Tree Star).

Bernelin-Cottet C., Deloizy C., Stanek O., Barc C., Bouguyon E., Urien C., Boulesteix O., Pezant J., Richard C.A., Moudjou M., Da Costa B., Jouneau L., Chevalier C., Leclerc C., Sebo P., Bertho N, & Schwartz-Cornil I. (2016). A Universal Influenza Vaccine Can Lead to Disease Exacerbation or Viral Control Depending on Delivery Strategies. Frontiers in Immunology, 7, 641.

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Mitochondrial Staining of Splenic B Cells

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After various days of activation, splenic B cells were washed with prewarmed MACS buffer (PBS, pH 7.2, supplemented with 0.5% BSA and 2 mM EDTA) and 1 × 10⁶ cells were resuspended in 100 μL of staining buffer containing MitoTracker Green (20 nM; M7514; Invitrogen) and MitoTracker DeepRed (20 nM; M22426; Invitrogen), and were incubated at 37°C for 10 min. Stained cells were washed with 1 mL of prewarmed MACS buffer and kept on ice to quench further staining. Samples were analyzed by flow cytometry within 30 min. In some experiments, cells were first stained with anti-IgG1 (PE Rat Anti-Mouse IgG1, Clone A85-1, Cat # 550083; BD Bioscience) anti-CD138 (BV421 Rat Anti-Mouse CD138, Clone 281–2, Cat #562610; BD Bioscience), or anti-CD69 (APC Hamster anti-mouse CD69 clone H1.2F3; BD Bioscience) antibodies for 30 min followed by MitoTracker dyes. Other antibodies used and analyses of germinal center cells are described in our previously published work [4 (link)]. Data were collected on a BD LSR II or Fortessa flow cytometer and analyzed with FlowJo software (Tree Star). Exclusion by Zombie Aqua™ (Biolegend) or DAPI was used to identify live cells and doublets were excluded by forward and side scatter height versus width analysis.

Nandi S., Liang G., Sindhava V., Angireddy R., Basu A., Banerjee S., Hodawadekar S., Zhang Y., Avadhani N.G., Sen R, & Atchison M.L. (2020). YY1 control of mitochondrial-related genes does not account for regulation of immunoglobulin class switch recombination in mice. European journal of immunology, 50(6), 822-838.

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Multi-parameter flow cytometry analysis of T cells

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Blood was collected by retro-orbital puncture. Spleen cells were obtained by mechanical disruption. After Fc receptor blocking (clone 2.4G2, 1:1,000; BD Biosciences), cells were stained with anti-CD4 PerCP-Cy5.5 (clone RM4-5; 1:800; BD Biosciences) or CD4 BV785 (clone RM4-5; 1:800; BioLegend), anti-TCR-β APC (clone H57-597; 1:200; BD Biosciences) or PE (clone H57-597; 1:100; BD Biosciences), anti-NKp46 FITC (clone 29A1.4; 1:300; eBiosciences), anti-CD8 Alexa700 (clone 53-6.7; 1:1,600; BioLegend), anti-CD25 PE-Cy7 (clone PC61; 1:300; eBiosciences), anti-CD62L BV605 (clone MEL-14; 1:800; BioLegend) and anti-CD44 APC-Cy7 (clone IM7; 1:400; BD Biosciences) in PBS/3% FCS. Then anti-Foxp3 eFluor450 (clone FJK-16 s; 1:150; eBioscience) and Ki67-PE (clone B56; 1:50; BD Biosciences) or Ki67-Alexa647 (clone B56; 1:50; BD Biosciences) staining was performed using the eBioscience Foxp3 staining kit according to the manufacturer's instructions. Cells were acquired on a Fortessa flow cytometer and analysed with FlowJo software (TreeStar).

Pérol L., Lindner J.M., Caudana P., Nunez N.G., Baeyens A., Valle A., Sedlik C., Loirat D., Boyer O., Créange A., Cohen J.L., Rogner U.C., Yamanouchi J., Marchant M., Leber X.C., Scharenberg M., Gagnerault M.C., Mallone R., Battaglia M., Santamaria P., Hartemann A., Traggiai E, & Piaggio E. (2016). Loss of immune tolerance to IL-2 in type 1 diabetes. Nature Communications, 7, 13027.

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