Cellquest software version 3

Manufactured by BD

Sourced in United States

CellQuest software version 3.3 is a data acquisition and analysis software for flow cytometry. It provides the core functionality for configuring and operating flow cytometry instruments to collect and analyze cell population data.

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Lab products found in correlation

Facscalibur flow cytometer, by BD (13 mentions) Facscalibur, by BD (11 mentions) Facscan, by BD (10 mentions) Facscan flow cytometer, by BD (7 mentions) Facscalibur system facscan, by BD (3 mentions) Facsort, by BD (2 mentions) Anti cd3 percp, by BD (2 mentions) Propidium iodide, by Merck Group (2 mentions) Facscalibur ow cytometer, by BD (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Facscalibur system, by BD (2 mentions) Annexin 5 fitc, by BD (2 mentions) Facscanto 2 flow cytometer, by BD (2 mentions) Cd146 pe, by BD (1 mentions) Igg1 fitc, by BD (1 mentions) Cd90 pe, by Bio-Rad (1 mentions) Cd105 pe, by Bio-Rad (1 mentions) Cd31 fitc, by Bio-Rad (1 mentions) Cd271 pe, by Miltenyi Biotec (1 mentions) Igg1 pe, by Bio-Rad (1 mentions)

Spelling variants of this product

CellQuest software (5116 citations) CellQuest (816 citations) CellQuest 3.0 software (157 citations) CellQuest version 3.3 (32 citations)

90 protocols using cellquest software version 3

Phenotypic profiling of MSCs and ECs

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Phenotypic characterisation was performed on culture expanded MSCs at different passages and on cultured ECs at p4 using: CD31-FITC (#MCA1738F), CD105-PE (#MCA1557PE), CD90-PE (#MCA90PE) (all from Serotec, Kidlington, UK), CD73-PE (#550257), CD146-PE (#550315) (both from BD Pharmingen, Oxford, UK), and CD271-PE (#130-091-885, Miltenyi Biotec). The isotype controls were IgG1-FITC (#550616, BD Pharmingen) and IgG1-PE (#MCA928PE, Serotec). A total of 2×10
⁵ cells was stained with 5 μl FITC- or PE-conjugated antibodies, and dead cells were excluded using 2 μg/ml propidium iodide (PI, #P1304MP, Invitrogen). Cells were acquired using FACScan equipped with CellQuest software version 3.1 (BD Biosciences) and the proportions of the different fractions were calculated as a percentage of total live cells.

Kouroupis D., Churchman S.M., McGonagle D, & Jones E.A. (2014). The assessment of CD146-based cell sorting and telomere length analysis for establishing the identity of mesenchymal stem cells in human umbilical cord. F1000Research, 3, 126.

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Apoptosis Induction in A549 Cells

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Following treatment with 10 µg/ml gemcitabine and 10 µM sorafenib at 37°C for 48 h, A549 cells were collected using centrifugation at 800 × g at 4°C for 10 min. Then, cells were stained with annexin V-fluorescein isothiocyanate (FITC) and PI using the Annexin V-FITC Apoptosis Detection kit (Nanjing KeyGen Biotech Co., Ltd.). Briefly, the cells were incubated with Annexin V-FITC/PI at 37°C for 20 min, and apoptotic cells were subsequently analyzed using a FACSCalibur flow cytometer (BD Biosciences); Annexin V⁺/PI⁺ and Annexin V⁺/PI⁻ cells were considered to be apoptotic. A549 cells treated with 100 nM docetaxel at 37°C for 48 h were used as positive control. The data was analyzed using CellQuest software version 3.1 (BD Biosciences).

Jiang S., Wang R., Zhang X., Wu F., Li S, & Yuan Y. (2020). Combination treatment of gemcitabine and sorafenib exerts a synergistic inhibitory effect on non-small cell lung cancer in vitro and in vivo via the epithelial-to-mesenchymal transition process. Oncology Letters, 20(1), 346-356.

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Tumor Cell Apoptosis Assay in Rabbits

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The tumor tissue from rabbits in the control, ADM-NP and ADM-NMC groups were minced into 5 mm³ pieces at the central and edge of the tumor and were used to prepare the monoplast suspension. Flow cytometry was then used to determine tumor cell apoptosis. The FITC Annexin V/PI kit (Thermo Fisher Scientific, Inc.) was used, following the manufacturer’s instructions. A flow cytometry analyzer FACSCalibur with Cell Quest software version 3.1 (BD Biosciences, Franklin Lakes, NJ, U.S.A.) was used to detect cellular apoptosis.

Chen K, & Zhang L. (2019). Effect of drug-loaded microbubbles combined with ultrasound on the apoptosis of cancer cells and the expression of Bax and Bcl-2 in a rabbit VX2 liver tumor model. Bioscience Reports, 39(5), BSR20181144.

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Mitochondrial ROS Evaluation in Cells

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Cells were seeded at a density of 1×10⁴/well into a 6-well plate in a 37°C incubator. After 24 h, 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA; 10 µM; Sigma-Aldrich; Merck KGaA) was added into the wells and was incubated at 37°C for 30 min. The cells were washed in PBS three times (2–3 min each time) to remove the DCFH-DA dye. The evaluation of mitochondrial reactive oxygen species (ROS) was performed using mitoSOX dye (Molecular Probes, USA), which selectively targets the mitochondrial matrix and emits red fluorescence when oxidized by ROS. The cells were stained with 2 µM mitoSOX dye and incubated for 15 min at 37°C. The cells were subsequently suspended in PBS and analyzed by flow cytometry. ROS levels were measured by FACSCalibur with Cell Quest software version 3.1 (BD Biosciences, San Jose, CA, USA).

Wang L., Lin R., Guo L, & Hong M. (2018). Rosuvastatin relieves myocardial ischemia/reperfusion injury by upregulating PPAR-γ and UCP2. Molecular Medicine Reports, 18(1), 789-798.

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Quantitative Analysis of Cellular Uptake

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To quantitatively examine the cellular uptake level of RPM/siRNA-Cy5, fluorescence-activated cell sorting (FACS) analysis was used. HUVECs were incubated with different RPM/siRNA complexes (Table 1) for 6 hours at 37°C. The incubation volume for all the groups was 1 mL, and the final concentration of siRNA-Cy5 was 100 nM (5 μL, 20 μM). Then, the cells were washed with phosphate-buffered saline (PBS) three times to remove any extracellular siRNA-Cy5, and then trypsinized and suspended in 10% FBS/DMEM to inhibit the trypsin activity. Cells were collected and suspended in 400 μL PBS within flow tubes, and cell fluorescence was determined with the BD FACSCalibur CellSorting System (model number BD FACSCALIBUR) (BD Biosciences, San Jose, CA, USA). Approximately 2×10⁵ cells were scanned per sample using the fluorescence channel (equipped with a 660 nm band pass filter) of the FACS system. Data were obtained and analyzed using CellQuest software (version 3.1) (BD Biosciences).

Liu L., Liu X., Xu Q., Wu P., Zuo X., Zhang J., Deng H., Wu Z, & Ji A. (2014). Self-assembled nanoparticles based on the c(RGDfk) peptide for the delivery of siRNA targeting the VEGFR2 gene for tumor therapy. International Journal of Nanomedicine, 9, 3509-3526.

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Cell Cycle and Apoptosis Analysis by Flow Cytometry

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Cells (2×10⁵/2 ml) were seeded in 6-well plates. Cells were centrifuged after 48 h of transfection and fixed with 70% ice-cold ethanol at 4°C for 24 h. Next, cells were harvested and washed with cold PBS twice [containing 100 mg/ml RNase A (EN0531; Thermo Fisher Scientific, Inc.) and 50 mg/ml propidium iodide (PI)] in the dark for 30 min at 4°C. The cell cycle distribution was analyzed using a FACScan laser flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) equipped with CellQuest software version 3.1 (BD Biosciences). All experiments were performed in triplicate. Similarly, cells were transfected for 48 and 96 h, then harvested and suspended in medium at a concentration of 2×10⁵ cells/2 ml. Annexin V-fluorescein isothiocyanate (5 µl) and PI (5 µl) were added to the cells, followed by incubation in the dark at 4°C for 15 min. Subsequently, cell apoptosis was measured using flow cytometry.

He F., Cheng X.M, & Gu W.L. (2018). Effects of cullin 4B on the proliferation and invasion of human gastric cancer cells. Molecular Medicine Reports, 17(4), 4973-4980.

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Evaluating Apoptosis using Annexin V/PI

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To assess apoptosis, an Annexin V/PI apoptosis kit (MultiSciences Biotech, Co., Ltd., Hangzhou, China) was used, according to the manufacturer's instructions. Briefly, following incubation in 6-well plates with osteoinductive medium in the presence of various concentrations of ZA (0, 0.01, 0.1, 1, 10 or 100 µM) for 1, 4 and 7 days, the cultured MC3T3-E1 cells were gently resuspended in binding buffer and incubated for 5 min at room temperature in the dark with 5 µl Annexin V-FITC and 10 µl PI. The AnnexinV-FITC and PI-labelled cells were analyzed using a flow cytometer (FACSort; BD Biosciences, Burlington, MA, USA). Using flow cytometry, dot plots of Annexin V-FITC, on the X-axis, against PI, on the Y-axis, were used to distinguish viable cells, which are negative for PI and Annexin V-FITC, early apoptotic cells (Annexin V-positive/PI-negative) and late apoptotic or necrotic cells (AnnexinV-FITC-positive/PI-positive staining). The resultant data was analyzed using CellQuest software version 3.1 (BD Biosciences, San Jose, CA, USA).

HUANG X., HUANG S., GUO F., XU F., CHENG P., YE Y., DONG Y., XIANG W, & CHEN A. (2015). Dose-dependent inhibitory effects of zoledronic acid on osteoblast viability and function in vitro. Molecular Medicine Reports, 13(1), 613-622.

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Flow Cytometry of T-Cell Subsets

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Peripheral blood T-cell subsets were measured by flow cytometry at diagnosis, using the following directly labeled antibodies: anti-CD3-PerCP, anti-CD4-FITC, anti-CD8-PE and anti-CD25-PE (BD Biosciences, San Jose, CA, USA). Data acquisition and analysis were carried out on a FACSCalibur flow cytometer, with CellQuest software version 3.1 (BD Biosciences).

Wang H., Tu M., Fu R., Wu Y., Liu H., Xing L, & Shao Z. (2014). The Clinical and Immune Characteristics of Patients with Hepatitis-Associated Aplastic Anemia in China. PLoS ONE, 9(5), e98142.

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Characterization of Hepatic Immune Cells

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Hepatic immune cells obtained from liver were stained with CD11b and Ly6G antibodies (eBioscience) to identify neutrophils or CD3 and NK1.1 antibodies (eBioscience) to identify iNKT cells. They were then subjected to flow cytometric analysis with the use of a FACS Calibur (BD Biosciences, San Jose, CA). Data were analyzed with CellQuest software version 3.1 (BD Biosciences) or FlowJo software version 7/8 (TreeStar Inc., Ashland, OR).

Kim J.W., Roh Y.S., Jeong H., Yi H.K., Lee M.H., Lim C.W, & Kim B. (2018). Spliceosome-Associated Protein 130 Exacerbates Alcohol-Induced Liver Injury by Inducing NLRP3 Inflammasome-Mediated IL-1β in Mice. The American journal of pathology, 188(4).

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Hepatic NPC Apoptosis Profiling

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Hepatic NPCs obtained from livers were stained with fluorescein isothiocyanate anti-CD4 antibody and allophycocyanin anti-CD25 antibody (e-Biosciences, San Diego, CA) on ice for 30 minutes followed by fixation with permeabilization concentrate buffer (e-Biosciences) at 4 C for 6 hours. After fixation, cells were washed twice with permeabilization buffer and stained with phosphatidylethanolamine anti-Foxp3 antibody in permeabilization buffer at room temperature for 30 minutes. For detection of early and late apoptosis, the liver NPCs were stained with anti-CD4 antibody and anti-CD25 antibody and washed with phosphate-buffered saline. Then, cells were suspended in binding buffer and supplemented with annexin V (AV) and propidium iodide (PI) for 15 minutes at room temperature without light exposure. The cells with fluorescence AV þ /PI À were considered to be early apoptotic cells, and those with fluorescence AV þ /PI þ were considered to be late apoptotic cells. Data were analyzed using CellQuest software version 3.1 (BD Biosciences, Franklin Lakes, NJ) or FlowJo software version 7.8 (FlowJo, Ashland, OR).

Roh Y.S., Kim J.W., Park S., Shon C., Kim S., Eo S.K., Kwon J.K., Lim C.W, & Kim B. (2018). Toll-Like Receptor-7 Signaling Promotes Nonalcoholic Steatohepatitis by Inhibiting Regulatory T Cells in Mice. The American journal of pathology, 188(11).

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