Facsdiva data analysis software

Manufactured by BD

Sourced in United States

FACSDiva is a data analysis software developed by BD (Becton, Dickinson and Company) for flow cytometry applications. The software provides tools for data acquisition, analysis, and visualization of cell populations based on their physical and fluorescent characteristics.

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Lab products found in correlation

Collagenase type 1, by Merck Group (3 mentions) 40 μm cell strainer cap, by BD (3 mentions) Anti mouse cd3 fitc or percp clone 145 2c11, by Miltenyi Biotec (2 mentions) Anti mouse foxp3 apc clone mf23, by BD (2 mentions) Facsaria, by BD (2 mentions) Percp conjugated anti cd45 clone 30 f11, by BioLegend (2 mentions) Alexa fluor700 gr 1 clone rb6 8c5, by BioLegend (2 mentions) Facsaria cytometer, by BD (1 mentions)

Close spelling variants of this product

FACSDiva software (5426 citations) FACSDiva (1830 citations) FACSDiva analysis software (32 citations)

7 protocols using facsdiva data analysis software

Purification of GFP-Expressing OBSCs

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OBSCs were plated at a density of 20,000–25,000 cells/cm² and, after 2 days in culture, neurospheres were disaggregated and analyzed by flow cytometry gating of propidium iodide-negative cells to determine the proportion of OBSCs expressing GFP. A FACSAria cytometer (BD Biosciences, San Diego, CA) equipped with a double Argon (488 nm) and Helium-Neon laser (633 nm) was used. Data were collected using a linear digital signal process. The emission filters used were BP 530/30 for GFP (FL1) and BP 616/23 for Propidium Iodide (FL3).
Cells expressing GFP were separated and purified by fluorescent activated cell sorting (FACS) using 20 psi pressure and a 100 μm nozzle aperture. GFP⁺ cells were collected directly in culture medium in autoclaved 5 mL polystyrene tubes. The cells were immediately counted and incubated in Trizol reagent to extract RNA or seeded in cell culture (see below). The purity of the isolated cells was confirmed by flow cytometry. Debris, dead cells and duplets were always excluded from the analysis.
Data were analyzed with FACSDiva data analysis software (BD Biosciences) and displayed using biexponential scaling.

Vergaño-Vera E., Díaz-Guerra E., Rodríguez-Traver E., Méndez-Gómez H.R., Solís Ó., Pignatelli J., Pickel J., Lee S.H., Moratalla R, & Vicario-Abejón C. (2014). Nurr1 Blocks the Mitogenic Effect of FGF-2 and EGF, Inducing Olfactory Bulb Neural Stem Cells to Adopt Dopaminergic and Dopaminergic-GABAergic Neuronal Phenotypes. Developmental neurobiology, 75(8), 823-841.

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Multiparametric Flow Cytometry of T Cells

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Multi-parameter flow cytometry was used to determine T cell subpopulations and their activation status. Cells were stained with anti-mouse CD3-FITC or -PerCP (clone 145-2C11, Miltenyi Biotec), CD4-PacificBlue (clone RM4-5), CD8-APC-Cy7 (clone 53-6.7), CD69 – PE (clone H1.2F3) and anti-mouse Foxp3-APC (clone MF23) (BD Biosciences). All antibodies were diluted according to manufacturers’ instructions. Gating was set using an isotype-matched control antibody (Supplementary Figure 2). Stained cells were analyzed on a LSR II flow cytometer using the FACSDiva data analysis software (BD Biosciences).

Budiu R.A., Vlad A.M., Nazario L., Bathula C., Cooper K.L., Edmed J., Thaker P.H., Urban J., Kalinski P., Lee A.V., Elishaev E.L., Conrads T.P, & Flint M.S. (2016). Restraint and Social Isolation Stressors Differentially Regulate Adaptive Immunity and Tumor Angiogenesis in a Breast Cancer Mouse Model. Cancer and clinical oncology, 6(1), 12-24.

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Corneal Cell Isolation and Flow Cytometry

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Cells were isolated from corneas as previously described [18 (link)]. Briefly, corneas were excised at 18 and 23 dpi and incubated in PBS-EDTA at 37°C for 15 minutes at 37°C. Stromas were separated from overlying epithelium and digested in 84 U collagenase type 1 (Sigma-Aldrich, St. Louis, MO) per cornea for 2 hours at 37°C and then were triturated to form a single-cell suspension. Suspensions were filtered through a 40-μm cell strainer cap (BD Labware, Bedford, MA) and washed and then stained. Suspensions were stained with PerCP-conjugated anti-CD45 (30-F11) and Alexa Fluor700-Gr-1 (RB6-8C5) (from BioLegend, San Diego, CA); FITC conjugated anti-CD4 (RM4-5), PE-conjugated anti-CD8α (53–6.7), PE-Cy7-conjugated anti-CD11c (HL3) (all BD PharMingen); eFluor450-conjugated CD11b (M1/70) (from eBiosciences, San Diego, CA). Cells were then analyzed on a flow cytometer (FACSAria with FACSDIVA data analysis software; BD Biosciences).

Yin X.T., Keadle T.L., Hard J., Herndon J., Potter C.A., Del Rosso C.R., Ferguson T.A, & Stuart P.M. (2015). Impaired Fas-Fas Ligand Interactions Result in Greater Recurrent Herpetic Stromal Keratitis in Mice. Journal of Immunology Research, 2015, 435140.

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Corneal Cell Isolation and Analysis

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Cells were isolated from corneas as previously described (21 (link)). Briefly, corneas were excised at 18 and 23 dpi and incubated in PBS-EDTA at 37°C for 15 minutes at 37°C. Stromas were separated from overlying epithelium and digested in 84 U collagenase type 1 (Sigma-Aldrich, St. Louis, MO) per cornea for 2 hours at 37°C and then were triturated to form a single-cell suspension. Suspensions were filtered through a 40-μm cell strainer cap (BD Labware, Bedford, MA) and washed and then stained. Suspensions were stained with: PerCP-conjugated anti-CD45 (clone 30-F11) and Alexa Fluor700-Gr-1 (clone RB6-8C5) (from BioLegend, San Diego, CA); FITC conjugated anti-CD4 (clone RM4–5), PE-conjugated anti-CD8α (clone 53–6.7), PE-Cy7-conjugated anti-CD11c (clone HL3), (all BD PharMingen); eFluor450-conjugated CD11b (clone M1/70) (from eBiosciences, San Diego, CA). Cells were then analyzed on a flow cytometer (FACSAria with FACSDIVA data analysis software; BD Biosciences).

West D.M., Del Rosso C.R., Yin X.T, & Stuart P.M. (2014). CXCL1, but not IL-6 is required for recurrent herpetic stromal keratitis. Journal of immunology (Baltimore, Md. : 1950), 192(4), 1762-1767.

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Isolation and Characterization of Corneal Immune Cells

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Cells were isolated from corneas as previously described (21 (link)). Briefly, corneas were excised at 18 and 23 dpi and incubated in PBS-EDTA at 37°C for 15 minutes at 37°C. Stromas were separated from overlying epithelium and digested in 84 U collagenase type 1 (Sigma-Aldrich, St. Louis, MO) per cornea for 2 hours at 37°C and then were triturated to form a single-cell suspension. Suspensions were filtered through a 40-μm cell strainer cap (BD Labware, Bedford, MA) and washed and then stained. Suspensions were stained with: PerCP-conjugated anti-CD45 (clone 30-F11), Alexa Fluor700-Gr-1 (clone RB6-8C5) and APC F4/80 (clone BM8) (from BioLegend, San Diego, CA); FITC conjugated anti-CD4 (clone RM4–5), PE-conjugated anti-CD8α (clone 53–6.7), PE-Cy7-conjugated anti-CD11c (clone N418), (all BD PharMingen); eFlour 450 anti-NK1.1 (clone PK136), PE-conjugated CD11b (clone M1/70) (from eBiosciences, San Diego, CA). The strategy for analysis was to initially gate on live cells and then the CD45⁺ cells. These CD45⁺ cells were further evaluated for T cell markers CD4 and CD8, or for macrophage markers F4/80⁺CD11b⁺GR-1⁻, or neutrophil markers GR-1⁺,CD11b⁺F4/80⁻, or dendritic cell marker CD11c⁺,F4/80⁻. Cells were then analyzed on a flow cytometer (FACSAria with FACSDIVA data analysis software; BD Biosciences.

Tajfirouz D., West D.M., Yin X.T., Potter C.A., Klein R, & Stuart P.M. (2017). CXCL9 compensates for the absence of CXCL10 during recurrent Herpetic Stromal Keratitis. Virology, 506, 7-13.

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Cell Cycle Analysis by Flow Cytometry

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Cells were treated with paclitaxel and stress hormones as described above. Briefly, cells were trypsinised, washed with PBS and fixed with 100% ethanol at 4 °C for 1 h. Cell suspensions were then incubated with 50 ng μl⁻¹ of RNase A for 15 min at 37 °C followed by 50 ng μl⁻¹ of propidium iodide for 30 min at 4 °C in the dark. Cells were analysed on a LSR II flow cytometer using the FACSDiva data analysis software (BD Biosciences, San Jose, CA, USA). Cell cycle distributions were generated using BD FACS Diva software version 6.1.1.

Reeder A., Attar M., Nazario L., Bathula C., Zhang A., Hochbaum D., Roy E., Cooper K.L., Oesterreich S., Davidson N.E., Neumann C.A, & Flint M.S. (2015). Stress hormones reduce the efficacy of paclitaxel in triple negative breast cancer through induction of DNA damage. British Journal of Cancer, 112(9), 1461-1470.

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Multiparametric Flow Cytometry of T Cells

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