Cytexpert v2

Manufactured by Beckman Coulter

Sourced in United States, Germany

The CytExpert v2.3 software is a data analysis tool designed for use with Beckman Coulter flow cytometry instruments. It provides users with the ability to analyze and interpret data generated from flow cytometry experiments.

Automatically generated - may contain errors

Lab products found in correlation

Cytoflex flow cytometer, by Beckman Coulter (17 mentions) Cytoflex, by Beckman Coulter (6 mentions) Anti brdu primary antibody, by Agilent Technologies (4 mentions) Cytoflex s, by Beckman Coulter (4 mentions) Alexa 488 conjugated anti mouse antibody, by Thermo Fisher Scientific (3 mentions) Propidium iodide, by Merck Group (3 mentions) Cytoflex lx flow cytometer, by Beckman Coulter (3 mentions) Cytoflex lx, by Beckman Coulter (3 mentions) Fluorescence activated cell sorting (facs), by BD (2 mentions) Fitc annexin 5 apoptosis detection kit 2, by BD (2 mentions) Annexin 5 fitc pi apoptosis detection kit, by BD (2 mentions) Cytoflex flow cytometry, by Beckman Coulter (2 mentions) Annexin 5 fitc apoptosis detection kit, by Elabscience (2 mentions) Rnase a, by Merck Group (2 mentions) Cd34 pe vio770 clone ac136, by Miltenyi Biotec (1 mentions) Cd73 pe clone rea804, by Miltenyi Biotec (1 mentions) Cd90 fitc clone rea897, by Miltenyi Biotec (1 mentions) Cd45 pe vio770 clone rea747, by Miltenyi Biotec (1 mentions) Synergy h1, by Agilent Technologies (1 mentions) Kaluza v1, by Beckman Coulter (1 mentions)

50 protocols using cytexpert v2

Cell Cycle and Apoptosis Analysis

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For cell cycle analysis, adherent cells seeded in six-well plates were treated with control or TRF paradigm for 2 days, and then, the cells were collected, washed with precooled PBS, and fixed with precooled 75% ethanol. Cell cycle analysis was conducted by propidium iodide (PI) staining according to the manufacturer’s protocols (P4170, Sigma Aldrich, St Louis, USA). Cell cycle assessments were performed with a CytoFLEX flow cytometer (Beckman Coulter, CA, USA) and CytExpert V2.3.0.84 software (Beckman Coulter, USA). Three replicates for each sample were detected.
For apoptosis analysis, adherent cells treated with control feeding or TRF for 2 days were collected, washed, and resuspended in precooled PBS. FITC Annexin V Apoptosis Detection Kit I (E-CK-A211, E-CK-A252, Elabscience, Wuhan, China) was used according to the manufacturer’s instructions. Annexin V staining was used to identify early apoptosis, while PI and DAPI staining were used to identify late apoptosis. Cell apoptosis assessments were performed with a CytoFLEX flow cytometer (Beckman Coulter, CA, USA) and CytExpert V2.3.0.84 software (Beckman Coulter, USA). Three replicates for each sample were detected.

Shi D., Fang G., Chen Q., Li J., Ruan X, & Lian X. (2023). Six-hour time-restricted feeding inhibits lung cancer progression and reshapes circadian metabolism. BMC Medicine, 21, 417.

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Flow Cytometric Immunophenotyping of MSCs

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Flow cytometry was used to score positive or negative MSC or hemato/endothelial markers (antibodies: CD34-PE Vio770 clone AC136, CD73-PE clone REA804, CD90-FITC clone REA897, CD105-PerCP Vio700 clone REA794 and CD45-PE Vio770 clone REA747; Miltenyi Biotec, Bergisch Gladbach, Germany. CD44-PerCP clone 44PP2; Immunostep, Salamanca, Spain) with a CytoFLEX flow cytometer (Beckman Coulter, Fullerton, CA, USA), collecting a minimum of 30,000 events. CytExpert v2.3 (Beckman Coulter) software was used to process data. Antibodies were used in the following combinations: CD73/90/105/45 and CD34/44.

Ragni E., Piccolo S., Taiana M., Visconte C., Grieco G, & de Girolamo L. (2024). Inflammation and Starvation Affect Housekeeping Gene Stability in Adipose Mesenchymal Stromal Cells. Current Issues in Molecular Biology, 46(1), 842-855.

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Evaluating Artificial QS and PSD Systems

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To evaluate the artificial QS system and the PSD system, the sfGFP gene was harnessed as the reporter. At the population level, the sfGFP output was measured by the microplate reader (Synergy H1, BioTek Co.) (excitation at 485 nm; emission at 528 nm). The single-cell fluorescence analysis was performed by the flow cytometer (CytoFLEX, Beckman Coulter, Inc.). The single-cell fluorescence analysis (excitation at 488 nm, emission at 525 nm) was performed by the flow cytometer (CytoFLEX, Beckman Coulter Inc.). Approximately 50,000 cells per sample were detected and analyzed by CytExpert (v2.3) software (Beckman Coulter, Inc.).

Li F.H., Tang Q., Fan Y.Y., Li Y., Li J., Wu J.H., Luo C.F., Sun H., Li W.W, & Yu H.Q. (2020). Developing a population-state decision system for intelligently reprogramming extracellular electron transfer in Shewanella oneidensis. Proceedings of the National Academy of Sciences of the United States of America, 117(37), 23001-23010.

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Single-Cell Immune Profiling of Endometrium

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The method of human endometrium or mouse uterine being dissected and digested into single cells was the same as that for single‐cell RNA sequencing above. Then, the released cells were resuspended in FACS staining buffer (0.1% BSA in PBS). To examine immune cell subsets, cells were stained for viability (FVS510) and the indicated antibodies for 15 min. Data were acquired using CytoFLEX (Beckman Coulter, USA) and analysis was performed using CytExpert v2.3 (Beckman Coulter, USA). The antibodies used were shown in Appendix Table S6.

Lv H., Sun H., Wang L., Yao S., Liu D., Zhang X., Pei Z., Zhou J., Wang H., Dai J., Yan G., Ding L., Wang Z., Cao C., Zhao G, & Hu Y. (2023). Targeting CD301 + macrophages inhibits endometrial fibrosis and improves pregnancy outcome. EMBO Molecular Medicine, 15(9), e17601.

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Flow Cytometry Data Analysis Workflow

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All flow cytometry data were analyzed using CytExpert v2.3 and Kaluza v1.5 (Beckman Coulter, Brea, CA). First, the compensation matrix was fine-tuned for the samples, if applicable. Next, the population gates were adjusted for each sample, a statistics table was prepared, and the population data were then exported to prepare charts and graphs using Excel 2013 (Microsoft, Redmond, WA). DLS data were analyzed using DelsaMax Software v1.6.1.17 (Beckman Coulter, Brea, CA).

Brittain GC I.V., Chen Y.Q., Martinez E., Tang V.A., Renner T.M., Langlois M.A, & Gulnik S. (2019). A Novel Semiconductor-Based Flow Cytometer with Enhanced Light-Scatter Sensitivity for the Analysis of Biological Nanoparticles. Scientific Reports, 9, 16039.

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Cell Cycle Analysis by Flow Cytometry

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Cells were grown to 70–80% confluency and were treated or transduced as indicated in the figures. After treatment, cells were harvested and washed with cold PBS before being fixed in 70% ethanol for two hours on ice. Cells were stained with PI/RNase solution (Invitrogen, F10797) for fifteen minutes before being analyzed via flow cytometry. Data was collected with Beckman Coulter CytExpert v2.3. Data was analyzed in FlowJo v10.8.1. First, debris was removed via gating under FSC-A/SSC-A, singlets were gated with FSC-A/FSC-H, and then cell cycle phases were determined using the Watson (Pragmatic) model (Supplementary Fig. 7).

Mittal K., Cooper G.W., Lee B.P., Su Y., Skinner K.T., Shim J., Jonus H.C., Kim W.J., Doshi M., Almanza D., Kynnap B.D., Christie A.L., Yang X., Cowley G.S., Leeper B.A., Morton C.L., Dwivedi B., Lawrence T., Rupji M., Keskula P., Meyer S., Clinton C.M., Bhasin M., Crompton B.D., Tseng Y.Y., Boehm J.S., Ligon K.L., Root D.E., Murphy A.J., Weinstock D.M., Gokhale P.C., Spangle J.M., Rivera M.N., Mullen E.A., Stegmaier K., Goldsmith K.C., Hahn W.C, & Hong A.L. (2024). Targeting TRIP13 in favorable histology Wilms tumor with nuclear export inhibitors synergizes with doxorubicin. Communications Biology, 7, 426.

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Apoptosis Quantification by Flow Cytometry

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The cells were harvested after intervention, washed twice with PBS, and then resuspended in 500 mL of PBS. The cells were transferred to 1.5 mL EP tubes, stained using the FITC Annexin V Apoptosis Detection kit II (BD Biosciences (San Jose, CA, USA), cat. No. 556570), and incubated in the dark for 10 min at room temperature, according to the manufacturer’s protocol. Subsequently, 10 µL of propidium iodide (PI, Beyotime Institute of Biotechnology) was added to the stained cells, and apoptotic cells were distinguished using a fluorescence-activated cell sorting analyzer (FACS; BD Biosciences). The results were analyzed with the Cytexpert V2.3 (Beckman Coulter, Inc., Brea, CA, USA) software. The apoptotic rate was calculated with the percentage of early + late apoptotic cells.

Sun Y., Liu X., Tong H., Yin H., Li T., Zhu J., Chen J., Wu L., Zhang X., Gou X, & He W. (2023). SIRT1 Promotes Cisplatin Resistance in Bladder Cancer via Beclin1 Deacetylation-Mediated Autophagy. Cancers, 16(1), 125.

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Annexin V-FITC/PI Apoptosis Assay

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Cellular apoptosis was analyzed using an Annexin V-FITC/PI Apoptosis Detection kit (BD Biosciences), according to the manufacturer's protocol. Briefly, cells were incubated with the indicated concentrations of osimertinib, ABT-263, osimertinib and ABT-263, or DMSO as the negative control for 24 h. The cells were subsequently trypsinized, washed and resuspended in 200 µl binding buffer, then stained with 5 µl PE and FITC for 15 min in the dark at room temperature. Finally, the stained cells were subjected to analysis by flow cytometry (Cytomics FC 500 MPL; Beckman Coulter, Inc.). The obtained data were analyzed by CytExpert v2.3 (Beckman Coulter, Inc.). The early apoptosis was evaluated based on the percentage of cells with Annexin V⁺/PI⁻, while the late apoptosis was that of cells with Annexin V⁺/PI⁺. The results are presented as the mean ± SD of three independent experiments.

Lu Y., Bian D., Zhang X., Zhang H, & Zhu Z. (2020). Inhibition of Bcl-2 and Bcl-xL overcomes the resistance to the third-generation EGFR tyrosine kinase inhibitor osimertinib in non-small cell lung cancer. Molecular Medicine Reports, 23(1), 48.

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Cisplatin-Induced Apoptosis in Bladder Cancer

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siRNA-transfected UMUC-3 and T24 cells were seeded into 6-well plates and incubated with cisplatin (0, 5 or 10 µg/ml) for 48 h. After washing twice with PBS, cells (~1×10⁶/ml) were resuspended in 500 ml PBS. Cells were transferred to 1.5 ml Eppendorf tubes and stained using the FITC Annexin V Apoptosis Detection kit II (BD Biosciences, cat. no. 556570) and incubated in the dark for 10 min at room temperature according to the manufacturer's protocol. Subsequently, 10 µl propidium iodide (PI, Beyotime Institute of Biotechnology) was added to the stained cells and apoptotic cells were distinguished by a fluorescence-activated cell sorting analyzer (FACS; BD Biosciences). The results were analyzed by Cytexpert V2.3 (Beckman Coulter, Inc.) software. The apoptotic rate was calculated by the percentage of early + late apoptotic cells.

Qin Z., Tong H., Li T., Cao H., Zhu J., Yin S, & He W. (2021). SPHK1 contributes to cisplatin resistance in bladder cancer cells via the NONO/STAT3 axis. International Journal of Molecular Medicine, 48(5), 204.

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Apoptosis and Cell Cycle Analysis via Flow Cytometry

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An annexin V/FITC and PI apoptosis detection kit and flow cytometry were adopted to evaluate cell apoptosis. The transfection of cells with shRNA was performed, which was followed by three washes with PBS. An Annexin V FITC-PI apoptosis detection kit (BD Biosciences, NJ, USA) was adopted to perform apoptosis staining on basis of the producer's guideline. A CytoFLEX flow cytometer (Beckman Coulter, USA) was employed to analyze stained cells. CytExpert V2.3.0.84 software (Beckman Coulter, USA) was used for data acquisition. For the assessment of the cell cycle allocation, PBS was adopted to wash the cells transfected with shRNA twice, Then, the overnight fixing of cells was performed with 75% ethanol (absolute ethanol/PBS ratio = 3:1) at -4 °C. After 30-minute incubation with propidium iodide (PI; 100 μg/ml) solution at room temperature (RT) in the dark, a CytoFLEX flow cytometer (Beckman Coulter, USA) was employed to measure the cells. CytExpert V2.3.0.84 software was adopted for the analysis of the cell cycle distribution.

Zhang Z., Zhong L., Dan W., Chu X., Liu C., Luo X., Wan P., Liu Z., Lu Y., Wang X, & Liu B. (2022). ZFP91 promotes cell proliferation and inhibits cell apoptosis in AML via inhibiting the proteasome-dependent degradation of RIP1. International Journal of Medical Sciences, 19(2), 274-285.

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