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Lsr fortessa with hts

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The LSR Fortessa with HTS (High Throughput Sampler) is a flow cytometry system designed for a wide range of applications. It features a compact footprint, advanced optics, and the HTS module for automated sample handling. The core function of this product is to provide a robust and efficient platform for flow cytometry analysis.

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Lab products found in correlation

Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) 7 aminoactinomycin d 7 aad, by BD (1 mentions) Pe cy7 anti human cd24, by BioLegend (1 mentions) Anti human cd133 pe, by Miltenyi Biotec (1 mentions) Aldefluor expression, by STEMCELL (1 mentions) 7 aad, by BD (1 mentions) Cytometric bead array cba, by BD (1 mentions)

Spelling variants of this product

Fortessa (1232 citations) Fortessa LSR (45 citations) BD LSR-Fortessa-HTS (9 citations)

5 protocols using lsr fortessa with hts

1

Annexin V-FITC Apoptosis Assay

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Cell apoptosis was determined by using a BD PharMingen FITC Annexin V Apoptosis Detection Kit I. Briefly, cells were harvested and washed with PBS twice. The pellets were resuspended in 1x Binding Buffer and incubated for 15 minutes at room temperature with 3 μL of FITC Annexin V and 3 μL of Propidium Iodide in the dark. Then 400 μL of 1x Binding Buffer was added to each tube prior to analysis. Samples were analyzed with a flow cytometer (LSR Fortessa with HTS, BD Biosciences, NJ, USA) and Summit (Beckman Coulter Inc.; Fullerton, CA).
Huang D., Chowdhury S., Wang H., Savage S.R., Ivey R.G., Kennedy J.J., Whiteaker J.R., Lin C., Hou X., Oberg A.L., Larson M.C., Eskandari N., Delisi D.A., Gentile S., Huntoon C.J., Voytovich U.J., Shire Z.J., Yu Q., Gygi S.P., Hoofnagle A.N., Herbert Z.T., Lorentzen T.D., Calinawan A., Karnitz L.M., Weroha S.J., Kaufmann S.H., Zhang B., Wang P., Birrer M.J, & Paulovich A.G. (2021). Multiomic analysis identifies CPT1A as a potential therapeutic target in platinum-refractory, high-grade serous ovarian cancer. Cell Reports Medicine, 2(12), 100471.
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2

Formaldehyde-based Cell Fixation and DAPI Staining

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Cells were harvested and washed twice with cold PBS. The pellets were resuspended in PBS with 1% formaldehyde and 0.2% Trition X-100. Then, the cells were incubated in PBS with DAPI for 2 h. The samples were analyzed with a flow cytometer (LSR Fortessa with HTS, BD Biosciences, San Jose, CA, USA) and Summit (Beckman Coulter Inc.; Fullerton, Brea, CA, USA).
Eskandari N., Senyuk V., Moore J., Kalik Z., Luan Q., Papautsky I., Moshiri A., Bocchetta M., Salami S.A., Oryan S, & Gentile S. (2021). Molecular Activation of the Kv11.1 Channel Reprograms EMT in Colon Cancer by Inhibiting TGFβ Signaling via Activation of Calcineurin. Cancers, 13(23), 6025.
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3

Characterization of Tumor Cells and NK Cells

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Pacific Blue anti-CD45 (HI30), allophycocyanin anti–HLA-ABC (G46-2.6), Alexa Fluor 488 anti-CD3 (UCHT1), and 7-aminoactinomycin D (7-AAD) were purchased from BD Biosciences (San Jose, CA). PE-Cy7 CD24 (ML5), Pacific Blue CD44 (IM7), biotin anti-MICA (6D4), biotin anti-Fas (DX2), and allophycocyanin-Cy7 anti-CD56 (HCD56) were purchased from BioLegend (San Diego, CA). PE anti-CD133 (AC133) was purchased from Miltenyi Biotec (Auburn, CA). PE anti-DR5 (DJR2-4) was purchased from eBioscience (San Diego, CA). We used a linked streptavidin secondary Ab for detection of biotinylated anti-MICA/B and anti-Fas Abs. Live tumor cells were gated and separated from NK cells by discriminating side scatter (SSC)high/CD45−/7-AAD− events. All samples were acquired on an LSRFortessa with HTS (BD Biosciences, San Jose, CA) and analyzed with FlowJo software (Tree Star, Ashland, OR).
Ames E., Canter R.J., Grossenbacher S.K., Mac S., Chen M., Smith R.C., Hagino T., Perez-Cunningham J., Sckisel G.D., Urayama S., Monjazeb A.M., Fragoso R.C., Sayers T.J, & Murphy W.J. (2015). NK Cells Preferentially Target Tumor Cells with a Cancer Stem Cell Phenotype. Journal of immunology (Baltimore, Md. : 1950), 195(8), 4010-4019.
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4

Analyzing Cancer Stem Cell Markers

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ALDEFLUOR™ expression (STEMCELL Technologies, Vancouver, BC, Canada) was determined according to the manufacturers’ instructions using diethylaminobenzaldehyde (DEAB) to inhibit ALDH activity and to control for background fluorescence (Additional file 1: Figure S1). Pacific Blue anti-human CD45 (HI30) and 7-AAD were purchased from BD Biosciences (San Jose, CA). PE-Cy7 anti-human CD24 and Pacific Blue anti-human CD44 were purchased from BioLegend (San Diego, CA). PE anti-human CD133 was purchased from Miltenyi Biotec (Auburn, CA). All samples were acquired on an LSR Fortessa with HTS (BD Biosciences, San Jose, CA) and analyzed with FlowJo software (TreeStar, Ashland, OR).
Canter R.J., Ames E., Mac S., Grossenbacher S.K., Chen M., Li C.S., Borys D., Smith R.C., Tellez J., Sayers T.J., Monjazeb A.M, & Murphy W.J. (2014). Anti-proliferative but not anti-angiogenic tyrosine kinase inhibitors enrich for cancer stem cells in soft tissue sarcoma. BMC Cancer, 14, 756.
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5

Cytokine Quantification from PBMC Supernatant

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Interleukin (IL)-1β, IL-2, IL-6, IL-8 and tumor necrosis factor TNF from PBMC culture supernatant were quantified using a BD™ Cytometric Bead Array (CBA) (BD Biosciences). The CBA assays were performed according to the manufacturer’s instruction using a five laser Special Order LSRFortessa™ with HTS (BD Biosciences). Cytokine concentrations (ng/mL) were calculated based on the sample MFI compared to the cytokine standard curves. BD™ FCAP Array software version 3.0 was used for data analysis. Graphs and statistical analysis were produced using GraphPad Prism version 8 (GraphPad Software Inc.).
Cobos C., Bansal P.S., Wilson D.T., Jones L., Zhao G., Field M.A., Eichenberger R.M., Pickering D.A., Ryan R.Y., Ratnatunga C.N., Miles J.J., Ruscher R., Giacomin P.R., Navarro S., Loukas A, & Daly N.L. (2022). Peptides derived from hookworm anti-inflammatory proteins suppress inducible colitis in mice and inflammatory cytokine production by human cells. Frontiers in Medicine, 9, 934852.
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