Facsdiva v6

Manufactured by BD

Sourced in United States, Belgium

The FACSDiva v6.1.3 is a flow cytometry software that enables data acquisition, analysis, and sorting. It provides a user interface for configuring and controlling flow cytometry instruments.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (25 mentions) Facscanto 2, by BD (24 mentions) Facsaria 2, by BD (10 mentions) Lsr 2 flow cytometer, by BD (6 mentions) Lsrfortessa flow cytometer, by BD (5 mentions) Lsrii flow cytometer, by BD (3 mentions) Facsaria 3, by BD (3 mentions) Fixation buffer, by BioLegend (3 mentions) Phosphate buffered saline (pbs), by Merck Group (3 mentions) Lsr 2, by BD (3 mentions) Propidium iodide, by Merck Group (3 mentions) Cytofix cytoperm kit, by BD (3 mentions) 7 aad, by BD (3 mentions) Fcs express v4, by De Novo Software (3 mentions) Lsrfortessa cell analyzer flow cytometer, by BD (3 mentions) Facs lysing solution, by BD (3 mentions) Ideas software, by BD (2 mentions) Facsuite v 1, by BD (2 mentions) Cytometer setup and tracking beads, by BD (2 mentions) Compbead, by BD (2 mentions)

Close spelling variants of this product

BD FACSDIVA software (V6.1) BD FACSDiva Software v6.1.3 FACSDIVA V6. software program FACSDiva

153 protocols using facsdiva v6

Isolation and Characterization of NK and NKT Cells from RA Patients

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The peripheral blood or synovial uid of RA patients was collected to isolate MNCs as described above.
The sample was centrifuged, and the pellet was resuspended in PBS. PerCP anti-human CD38 and PE anti-human CD3 FITC/(CD16 + CD56) (10 µg per 10^7 cells in 500 µl volume) (1:1) were added, and the mixture was incubated for 30 min at 4 °C. The cells were washed, collected by centrifuging, and resuspended in PBS. The labeled cells were sorted using a BD FACS ARIA II with BD FACS Diva V6 software (BD Biosciences, USA). The CD38 and CD16 + CD56-positive and CD16 + CD56-negative gates were established, and the CD38 + NK and CD38 + NKT cells were differentiated by CD3+/-in the CD38 + and CD16 + CD56 + cell populations. The cell population in the negative gate was composed of MNCs depleted of CD38 + CD16 + CD56 + cells. The CD38 + NK cells contained a CD3-CD38 + CD16 + CD56 + population, and the CD38 + NKT cells contained a CD3 + CD38 + CD16 + CD56 + population. The sorted populations were collected into RPMI-1640 media (Gibco) containing 10% human heat-inactivated pooled AB serum (Gibco). The viability of the sorted cells was determined by trypan blue exclusion (Thermo Fisher Scienti c) by routine protocol. After sorting, the samples were tested for purity by a BD FACS ARIA II with BD FACS Diva V6 software (BD Biosciences, USA).

Wang H., Fang K., & Chang X. (2020). High Levels of CD38+ NK Cells Contribute to Immune Imbalance of Th1/Th2 and Th17/Treg in Rheumatoid Arthritis.

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Quantification of Immune Cell Subsets

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Quantification of T cells, B cells and plasmablasts was performed on 100 µl heparinized whole blood exactly as described before⁴ (link)
^,¹⁹ using monoclonal antibodies towards CD3 (clone SK7), CD19 (clone HIB19), CD27 (clone L128), CD38 (clone HB7), and IgD (clone IA6-2). Cells were analyzed using flow cytometry (FACS-Canto-II or FACSLyric) and FACS-Diva-V6.1.3 or FlowJo software (BD Biosciences). Among total lymphocytes, T and B cells were identified by expression of CD3 and CD19, respectively. Plasmablasts were gated as CD38 positive cells among IgD-CD27+ CD19 positive switched-memory B cells as described before.⁴ (link) Differential blood counts were used to calculate absolute lymphocyte numbers.

Schmidt T., Klemis V., Schub D., Schneitler S., Reichert M.C., Wilkens H., Sester U., Sester M, & Mihm J. (2022). Cellular immunity predominates over humoral immunity after homologous and heterologous mRNA and vector-based COVID-19 vaccine regimens in solid organ transplant recipients. American Journal of Transplantation, 21(12), 3990-4002.

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Quantifying Endothelial Cell Adhesion Markers

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At the basal state and after stimulations, non-permeabilized cells were detached by 5 mM EDTA, washed, and re-suspended in 0.5% BSA solution. The cells were treated and incubated with anti-VCAM-1 PE conjugate (Santa Cruz Biotechnology, sc-13160, 1:100) and with anti-ICAM-1 FITC conjugate (Santa Cruz Biotechnology, sc-107, 1:100) as previously described (Ucci et al., 2019 (link)). One test tube for the basal state and one for the TNF-α-treated condition were incubated with anti-VCAM-PE and anti-ICAM-FITC isotypes (normal mouse IgG2a FITC-conjugated and normal mouse IgG1 PE-conjugated, Santa Cruz Biotechnology, 1:100) as negative controls. Flow cytometry analysis was performed on a BD FACS Canto II flow cytometer (BD Bioscences) and for each sample 1 × 10⁴ events were analyzed using FACSDiva v 6.1.3 (BD Biosciences) and FlowJo 8.3.3 software (Tree Star Inc., Ashland, United States). All the results are expressed as mean fluorescence intensity (MFI) ratio ±standard deviation (SD). Each value was calculated by dividing the MFI of positive events by the MFI of negative events (MFI of secondary antibody). The experiment was carried out on four different strains for C-HUVECs and four different strains for GD-HUVECs, each in technical triplicate.

Pipino C., Bernabé-García Á., Cappellacci I., Stelling-Férez J., Di Tomo P., Santalucia M., Navalón C., Pandolfi A, & Nicolás F.J. (2022). Effect of the Human Amniotic Membrane on the Umbilical Vein Endothelial Cells of Gestational Diabetic Mothers: New Insight on Inflammation and Angiogenesis. Frontiers in Bioengineering and Biotechnology, 10, 854845.

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Optimizing Flow Cytometry Setup for EV and Liposome Analysis

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To broadly setup scatter parameters for EV and liposome detection, Megamix-Plus beads (Biocytex, Marseille, France) were acquired according to manufacturer’s instructions (threshold on FITC channel). Height (H) signals and logarithmic or bi-exponential modes were selected for all parameters.
Instrument performances were monitored by the Cytometer Setup and Tracking Module and further validated by the acquisition of Rainbow Beads (BD Biosciences, San Jose, CA, USA ) [38 (link),59 (link),60 (link)]. Data were analyzed using FACSDiva v 6.1.3 (BD Biosciences, San Jose, CA, USA), FACSuite v 1.0.6.5230 (BD Biosciences, San Jose, CA, USA) and FlowJo v 10.0.7 (TreeStar, now Becton, Dickinson and Company, Ashland, OR, USA) software.

Simeone P., Celia C., Bologna G., Ercolino E., Pierdomenico L., Cilurzo F., Grande R., Diomede F., Vespa S., Canonico B., Guescini M., Stocchi V., Lotti L.V., Guagnano M.T., Stellin L., Papa S., Trubiani O., Marchisio M., Miscia S, & Lanuti P. (2020). Diameters and Fluorescence Calibration for Extracellular Vesicle Analyses by Flow Cytometry. International Journal of Molecular Sciences, 21(21), 7885.

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Multimodal Data Acquisition Protocol

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FACS data acquisition were performed in FACSDiva v6.1.3 (BD) and analyzed in FlowJo v7.6.3 (FLOWJO LLC). Quantitative PCR data acquisition were performed in QuantStudio Real-Time PCR Software v1.3 (Applied Biosystems). Force measurements were acquired using LabChart v6.1.1 (ADInstruments). Immunostaining data were acquired using ZEN v2.3 pro (Zeiss). Fiber counting and measurements were performed with ImageJ v2.0.0-rc-65/1.52a (NIH).

Chan S.S., Arpke R.W., Filareto A., Xie N., Pappas M.P., Penaloza J.S., Perlingeiro R.C, & Kyba M. (2018). Skeletal muscle stem cells from PSC-derived teratomas have functional regenerative capacity. Cell stem cell, 23(1), 74-85.e6.

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Intracellular Phospho-Protein Analysis in CRC

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To observe STAT phosphorylation, single-cell suspensions of CRC tumor tissue and PBMC were stimulated with 10 ng IL-2, IL-6, and IL-10 for 15 min at 37 °C. For intracellular phospho-protein staining for flow cytometry, stimulated and unstimulated cells were fixed with a final concentration of 1.5% paraformaldehyde in culture medium for 10 min and subsequently pelleted. Thoroughly resuspended cells were permeabilized in 1 mL ice-cold MeOH per 10⁶ cells and stored at −70 °C for approximately two weeks before staining. Cells were washed three times with PBS containing 0.5% BSA, and resuspended in staining media at 0.5–1 × 10⁶ cells per 100 μL. Anti-phospho-STAT-1 (pY701), anti-phospho-STAT-3 (pY705), and anti-phospho-STAT-5 (pY694) antibodies (BD Biosciences, San Diego, CA, USA), as well as monoclonal antibodies against lineage markers, were added and incubated in the dark for 30 min. Cells were subsequently washed with staining media and pelleted. Finally, samples were resuspended in 200 μL staining media and analyzed by FACSCanto II using FACSDiva v6.1.3 (BD Biosciences, San Diego, CA, USA).

Yun J.W., Lee S., Kim H.M., Chun S., Engleman E.G., Kim H.C, & Kang E.S. (2019). A Novel Type of Blood Biomarker: Distinct Changes of Cytokine-Induced STAT Phosphorylation in Blood T Cells Between Colorectal Cancer Patients and Healthy Individuals. Cancers, 11(8), 1157.

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Protein Expression Quantification by Flow Cytometry

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Protein expression of ALP, OPN, OC, and COL1a1 was evaluated by flow cytometry analysis, following a previous protocol [35 (link)]. Samples were processed using a FACS Canto II flow cytometer (BD Biosciences, San Jose, CA, USA), and data were analyzed using FACSDiva v6.1.3, IDEAS software (BD Biosciences, San Jose, CA, USA), and FlowJo v8.3.3 software (Tree Star Inc., Ashland, OR, USA). Data are indicated as a mean fluorescence intensity (MFI) ratio calculated by dividing the MFI of positive events by the MFI of negative events (MFI of secondary antibody).

Pelusi L., Mandatori D., Di Pietrantonio N., Del Pizzo F., Di Tomo P., Di Pietro N., Buda R., Genovese S., Epifano F., Pandolfi A., Fiorito S, & Pipino C. (2022). Estrogen Receptor 1 (ESR1) and the Wnt/β-Catenin Pathway Mediate the Effect of the Coumarin Derivative Umbelliferon on Bone Mineralization. Nutrients, 14(15), 3209.

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Comparative Flow Cytometry Analysis of Anthracycline-Treated Cells

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Doxorubicin- and epirubicin-treated MDA-MB-231 cells were analyzed for their intrinsic fluorescence, acquiring 20,000 events/test using different flow cytometry platforms (FACSCanto II, FACSVerse—both from BD Biosciences; CytoFLEX—Beckman Coulter, Fullerton, CA, USA) as well as an AMINS ImageStream (Luminex Corporation, Austin, TX), equipped with a 488 nm solid-state laser (40 mW) and Inspire software (v 4.1.434.0) [6 (link)].
To obtain comparable results, flow cytometry analyses were standardized by daily running quality controls, including check-ups with Cytometer Setup and Tracking Beads (CS&T, BD). Debris and doublets were excluded from the analysis, and single events were analyzed for different purposes. Optimal photomultiplier (PMT) voltages were established for each channel [30 (link),31 (link)]. FACSDiva v 6.1.3 and FACSSuite v 1.0.6.5230 were used for data acquisition, and FlowJo v 10.8.1 Software (BD Biosciences) was used for data analysis.

Catitti G., De Fabritiis S., Brocco D., Simeone P., De Bellis D., Vespa S., Veschi S., De Lellis L., Tinari N., Verginelli F., Marchisio M., Cama A., Patruno A, & Lanuti P. (2022). Flow Cytometry Detection of Anthracycline-Treated Breast Cancer Cells: An Optimized Protocol. Current Issues in Molecular Biology, 45(1), 164-174.

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CRISPR-Cas9 Genetic Manipulation of ATG7

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TSA cells were transfected with a control commercial CRISPR-cas9 plasmid (#CRISPR06-1EA, from Millipore Sigma) or with a CRISPR-cas9 plasmid specific for Atg7 (custom-made by Millipore Sigma based on #CRISPR06-1EA), using the TransIT-CRISPR® reagent (#T1706, Sigma Aldrich). GFP⁺ clones were sorted on an FACSAria II Sorter operated by FACSDiva™ v. 6.1.3 (from BD Biosciences) into 96-well plates, followed by clone selection and confirmation of ATG7 status by immunoblotting.

Buqué A., Bloy N., Perez-Lanzón M., Iribarren K., Humeau J., Pol J.G., Levesque S., Mondragon L., Yamazaki T., Sato A., Aranda F., Durand S., Boissonnas A., Fucikova J., Senovilla L., Enot D., Hensler M., Kremer M., Stoll G., Hu Y., Massa C., Formenti S.C., Seliger B., Elemento O., Spisek R., André F., Zitvogel L., Delaloge S., Kroemer G, & Galluzzi L. (2020). Immunoprophylactic and immunotherapeutic control of hormone receptor-positive breast cancer. Nature Communications, 11, 3819.

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Quantifying Sub-G0 Cell Population

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To measure sub-G0 cell population, cells were trypsinized, washed with ice-cold PBS, and fixed in 70% ethanol at −20°C. Cells were washed with PBS and incubated with RNase 20 μg ml⁻¹ and propidium iodide (PI) 0.5 μg ml⁻¹ (Sigma-Aldrich). All analyses were performed with LSR II Flow Cytometer and FACSDiva v.6.1.3 software (BD Biosciences).

Wickström M., Dyberg C., Milosevic J., Einvik C., Calero R., Sveinbjörnsson B., Sandén E., Darabi A., Siesjö P., Kool M., Kogner P., Baryawno N, & Johnsen J.I. (2015). Wnt/β-catenin pathway regulates MGMT gene expression in cancer and inhibition of Wnt signalling prevents chemoresistance. Nature Communications, 6, 8904.

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