Cellquest pro 4

Manufactured by BD

Sourced in United States

CellQuest Pro 4.0.2 is a software application for flow cytometry data acquisition and analysis. It provides a user interface for controlling and configuring flow cytometry instruments, as well as tools for visualizing and analyzing the collected data.

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Lab products found in correlation

Facscalibur, by BD (21 mentions) Facscan flow cytometer, by BD (3 mentions) Propidium iodide, by Merck Group (2 mentions) Facs accuri, by BD (2 mentions) Alexa fluor 647 conjugated goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Facscan, by BD (2 mentions) Modfit lt 3, by Verity Software House (2 mentions) Metamorph software, by Molecular Devices (2 mentions) Facscalibur flow cytometer, by BD (2 mentions) Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions) Cell cycle and apoptosis analysis kit, by Keygen Biotech (1 mentions) Apoptag peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions) Cm h2dcfda, by Thermo Fisher Scientific (1 mentions) Facs calibur e6147 cytometer, by BD (1 mentions) Antibody diluent, by Agilent Technologies (1 mentions) Facsverse, by BD (1 mentions) Winmdi 2, by BD (1 mentions) Propidium iodide, by BD (1 mentions) Rnase a, by AppliChem (1 mentions) Alexa fluor 647 conjugated goat anti human igg, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CellQuest™ Pro Software 4.0.2 CellQuest Pro Version 4.0.2 software FACSCalibur with CellQuest Pro 4.0.2 CellQuest Pro Software 6.0.4 BD CellQuest Pro CellQuest

36 protocols using cellquest pro 4

Cell Viability and Cell Cycle Analysis

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DF-1 cells (4,000 cells/well) were cultured in 96-well plates and when the cell density reached 70-80%, transfection was performed. A total of 10 µl Cell Counting Kit-8 (CCK-8; Dojindo Molecular Technologies, Inc.) reagent was added to each well 24, 48 and 72 h after transfection and then incubated at 37˚C for 4 h according to the manufacturer's instructions. The absorbance was recorded at 450 nm.
The cell cycle assay was performed in 6-well plates and when the cell density reached 70-80%, transfection was performed. Flow cytometry was used to evaluate the distribution of cells in different phases of the cell cycle, basing the evaluation on the DNA content of 500 µl 1% propidium iodide (PI; Thermo Fisher Scientific, Inc.)-stained nuclei. After transfection for 48 h, cells were trypsinized and resuspended in 75% ethanol at a density of 1x10⁶ cells/ml, overnight at 4˚C, resuspended in PBS and dyed using the cell cycle dye solution (Cell Cycle and Apoptosis Analysis kit; Nanjing KeyGen Biotech Co., Ltd.) for 30 min at 37˚C in the dark. A flow cytometry (BD FACScalibur; BD Bioscience) was used to evaluate the cell cycle. Each group contained three replicate wells and all of the experiments were repeated independently three times. The results were analyzed using CellQuest Pro 4.0.2 (BD Biosciences).

Zhang X., Liao Z., Wu Y., Yan Y., Chen S., Lin S., Chen F, & Xie Q. (2020). gga-microRNA-375 negatively regulates the cell cycle and proliferation by targeting Yes-associated protein 1 in DF-1 cells. Experimental and Therapeutic Medicine, 20(1), 530-542.

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Cell Cycle and Apoptosis Analysis

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For cell cycle analysis, the cells were fixed with 70% ethanol in PBS at 4 °C for 30 min. After washing twice with PBS, the cells were stained with 40 μg/ml PI (Sigma-Aldrich) plus RNase for 30 min at room temperature. The cells were analyzed using flow cytometry (FACSCalibur; BD Bioscience) with excitation set at 488 nm and emission detected in the FL-2 channel (565–610 nm). The samples were analyzed using CellQuest Pro 4.0.2 software (BD Biosciences), and quantification was performed using FlowJo (Tree Star, Inc., Ashland, USA). Small cell debris was excluded by gating on a forward scatter plot. For PI staining, the levels of apoptosis were reported and gated as percentages of sub-G₁. To observe nuclear condensation, DAPI (Sigma-Aldrich)-stained cells were observed using a fluorescence microscope (BX51; Olympus, Tokyo, Japan). To detect apoptosis in osteoclast-like cells, the cells were fixed with 3.7% formaldehyde for 30 min. Then, TUNEL staining was performed using an ApopTag Peroxidase in situ Apoptosis Detection Kit (Millipore, Billerica, MA) according to the manufacturer’s instructions. Several randomly selected areas per specimen were analyzed.

Tai T.W., Chen C.Y., Su F.C., Tu Y.K., Tsai T.T., Lin C.F, & Jou I.M. (2017). Reactive oxygen species are required for zoledronic acid-induced apoptosis in osteoclast precursors and mature osteoclast-like cells. Scientific Reports, 7, 44245.

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Evaluation of Lysosomal and Mitochondrial Activity

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Cell cultures were vitally stained with acridine orange (AO; Sigma-Aldrich) at a concentration of 5 μg/ml for 15 min and then washed with PBS. AO is a fluorescent cationic dye that can be employed to measure the levels of acidic vesicular organelles (lysosomes) within cells. The levels of acidic vesicular organelles were analyzed using the FACSCalibur (BD Biosciences) with excitation set at 488 nm, and emission from fluorescent AO was detected by FL-3 channel (670 nm).
The mitochondrial membrane potential was determined using tetramethylrhodamine methyl ester (TMRM). Cell cultures were vitally stained with 0.5 μM TMRM for 30 min at 37°C and then washed with PBS. The fluorescence intensity of TMRM was analyzed using flow cytometry with FL-2 channel (564~606 nm). Samples were analyzed using CellQuest Pro 4.0.2 software (BD Biosciences) and quantification was performed using WinMDI 2.9 software (Scripps Research Institute, La Jolla, CA, USA).

Lai M.C., Chang C.M, & Sun H.S. (2016). Hypoxia Induces Autophagy through Translational Up-Regulation of Lysosomal Proteins in Human Colon Cancer Cells. PLoS ONE, 11(4), e0153627.

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Cell Cycle Analysis by Flow Cytometry

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The cell cycle was analyzed using propidium iodide (PI; Sigma-Aldrich) staining, and then, the cells were analyzed using flow cytometry (FACSCalibur; BD Biosciences, San Jose, CA) with excitation at 488 nm and emission detected in the FL-2 channel (565–610 nm). The percentages of cells were analyzed by using CellQuest Pro 4.0.2 software (BD Biosciences).

Hsieh T.H., Tsai T.T., Chen C.L., Shen T.J., Jhan M.K., Tseng P.C, & Lin C.F. (2020). Senescence in Monocytes Facilitates Dengue Virus Infection by Increasing Infectivity. Frontiers in Cellular and Infection Microbiology, 10, 375.

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Quantifying Cellular Oxidative Stress

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Cells were seeded at 7 × 10⁴/well in 24-well plates overnight. After GAS infection as described above, cells were collected and then coincubated with a cellular reactive oxygen species detection dye, carboxymethyl-H₂-dichlorofluorescein diacetate (CM-H₂DCFDA) (C6827; Invitrogen), at a concentration of 5 μM for 5 min at 37°C in the dark. After washing, cells were collected and analyzed using flow cytometry (FACSCalibur; BD Biosciences) with excitation set at 488 nm. The emission was detected with the FL-1 channel, followed by analysis with CellQuest Pro 4.0.2 software (BD Biosciences).

Cheng Y.L., Kuo C.F., Lu S.L., Hiroko O., Wu Y.N., Hsieh C.L., Noda T., Wu S.R., Anderson R., Lin C.F., Chen C.L., Wu J.J, & Lin Y.S. (2019). Group A Streptococcus Induces LAPosomes via SLO/β1 Integrin/NOX2/ROS Pathway in Endothelial Cells That Are Ineffective in Bacterial Killing and Suppress Xenophagy. mBio, 10(5), e02148-19.

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Cell Death Induction and Analysis

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Cells were treated with 125 μM of H₂O₂ for 16 hours or serum starvation for 5 days to induce cell death. Cell cycle of sub-G1 analysis was performed and quantified using the FACS-Calibur E6147 Cytometer and CellQuest Pro 4.02 software (BD Biosciences) as described previously [21 (link)].

Chiang K.C., Tsui K.H., Lin Y.H., Hou C.P., Chang K.S., Tsai H.H., Shin Y.S., Chen C.C., Feng T.H, & Juang H.H. (2019). Antioxidation and Antiapoptosis Characteristics of Heme Oxygenase-1 Enhance Tumorigenesis of Human Prostate Carcinoma Cells. Translational Oncology, 13(1), 102-112.

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Flow Cytometry Analysis of sYFP2 Expression

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Cell samples were withdrawn during the exponential phase (OD₆₀₀ of ~0.4) and at indicated time points, washed with 1× PBS, fixed with paraformaldehyde (4% in 1× PBS) for 30 min on ice, and stored at 4 °C until measurements. Flow cytometry experiments were performed with a FACSCalibur (BD Bioscience, San Jose, CA, USA) using the CellQuest Pro 4.0.2 (BD) software. Samples were acquired using the forward scatter (Amp: 10², Amp gain: 1.00), side scatter (500 V, Amp gain: 1.00) to exclude debris and a fluorescence detector FL1-H (excitation: 488 nm, emission: 530 nm, 500 V, Amp gain: 1.00) for the relative quantification of sYFP2 signals. Analysis was performed with normalized data sets (DownSample 2.0.0 plugin; 10,000 events) using FlowJo v. 10.6.2 (BD). R package ggplot2 (version 3.3.2) with function geom_density and count variables was used to draw smoothed distribution plots.

Edelmann D., Leinberger F.H., Schmid N.E., Oberpaul M., Schäberle T.F, & Berghoff B.A. (2021). Elevated Expression of Toxin TisB Protects Persister Cells against Ciprofloxacin but Enhances Susceptibility to Mitomycin C. Microorganisms, 9(5), 943.

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Flow Cytometric Analysis of Galectin-3 Expression

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To detect galectin-3 expression, we fixed, stained, and analyzed cells. For the flow cytometric analysis, cells were stained with anti-galectin-3 antibodies, followed by incubation with a mixture of Alexa Fluor 488-conjugated goat anti-rabbit IgG. Cells were analyzed using flow cytometry (FACSCalibur; BD Biosciences, San Jose, CA) with excitation at 488 nm; emission was detected with the FL-1 channel (at 515∼545 nm). Samples were analyzed using CellQuest Pro 4.0.2 software (BD Biosciences), and quantification was performed using WinMDI 2.8 software (The Scripps Institute, La Jolla, CA). Small cell debris was excluded by gating on a forward scatter plot. After washing twice with PBS, tissue sections were incubated with primary antibodies in antibody diluents (DAKO, Carpentaria, CA) at 4°C overnight.

Tseng P.C., Chen C.L., Shan Y.S, & Lin C.F. (2016). An increase in galectin-3 causes cellular unresponsiveness to IFN-γ-induced signal transduction and growth inhibition in gastric cancer cells. Oncotarget, 7(12), 15150-15160.

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Antibody Staining Dynamics in T Cells

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Infected T cells were immunostained on ice with the primary anti-HA antibody (clone 16B12; BioLegend). Cells were then shifted to 37°C for the indicated durations, placed back on ice, and immunostained on ice with secondary Alexa Fluor 647-conjugated goat anti-mouse IgG (Invitrogen), followed by PFA fixation. Flow cytometry analysis was performed using the FACSCalibur or FACS Accuri system with BD CellQuest Pro 4.0.2 software (BD Pharmingen) and FlowJo V10 software (FlowJo).

Passos V., Zillinger T., Casartelli N., Wachs A.S., Xu S., Malassa A., Steppich K., Schilling H., Franz S., Todt D., Steinmann E., Sutter K., Dittmer U., Bohne J., Schwartz O., Barchet W, & Goffinet C. (2019). Characterization of Endogenous SERINC5 Protein as Anti-HIV-1 Factor. Journal of Virology, 93(24), e01221-19.

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Cell Cycle Analysis of Irradiated B16F10 Cells

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The cell cycle distributions were analyzed using flow cytometry. B16F10 cells
were first irradiated with the indicated doses of X-rays and C-ion beams. At 6,
12, 24, 48, and 72 h post-radiation, the cells were dispersed using EDTA-free
trypsin, washed with phosphate-buffered saline (PBS), then fixed using
prechilled 75% ethanol, and stored at −20°C overnight. Fixed cells were
centrifuged, washed, and incubated in RNAse and propidium iodide prior to
measurement of DNA-content using a BD FACSVerse (BD Biosciences, USA). The
minimum of 10⁴ ungated cells were analyzed using BD CellQuest Pro
4.0.2 (BD, Biosciences, U.S.A.) and the cell cycle phases were evaluated using
ModFit LT 3.0 (Verity Software House, Inc., U.S.A.).

Li S., Huang H., Xing M., Qin J., Zhang H., Liu Y., Zhang L., Zhang C., Tian Z., Gao X., Zhao R, & Mao A. (2022). Carbon Ion Induces Cell Death and G2/M Arrest Through pRb/E2F1Chk2/Cdc2 Signaling Pathway in X-ray Resistant B16F10 Melanoma Cells. Dose-Response, 20(2), 15593258221092364.

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