Cytoflex lx

Manufactured by Beckman Coulter

Sourced in United States, Germany

The CytoFLEX LX is a flow cytometer designed for advanced research applications. It offers high-performance detection and analysis of cells and particles in a compact and user-friendly platform.

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Lab products found in correlation

Cytexpert software, by Beckman Coulter (13 mentions) Cytoflex s, by Beckman Coulter (9 mentions) Propidium iodide, by Thermo Fisher Scientific (9 mentions) Cytexpert, by Beckman Coulter (8 mentions) Lsrfortessa, by BD (8 mentions) Facscanto 2, by BD (7 mentions) Cd11a apc, by BioLegend (6 mentions) Cd8 pe cy7, by BD (6 mentions) Kaluza 2, by Beckman Coulter (6 mentions) Cytofix cytoperm kit, by BD (5 mentions) Tryple, by Thermo Fisher Scientific (5 mentions) Pd 1 fitc, by BioLegend (5 mentions) Cx3cr1 apc cy7, by BioLegend (5 mentions) Granzyme b percp, by Novus Biologicals (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (5 mentions) Cytexpert 2, by Beckman Coulter (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions) Bim pe, by Cell Signaling Technology (4 mentions) Zombie aqua fixable viability kit, by BioLegend (4 mentions)

Close spelling variants of this product

CytoFLEX LX cytometer CytoFLEX LX Analyzer CytoFLEX LX flow cytometer CytoFLEX LX Flow Cytometry System CytoFLEX LX flow cytometry CytoFLEX S/LX CytoFLEX LX FACS CYTOFLEX LX platform CytoFLEX LX device CytoFLEX LX system

476 protocols using cytoflex lx

Apoptosis and Cell Cycle Analysis

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Cell apoptosis analysis was performed using the Annexin V-APC/propidium iodide (PI) apoptosis detection kit (KGA1030-50, KeyGEN BioTECH, Nanjing, China). According to the manufacturer’s instructions, HCC cells were harvested, washed with PBS, resuspended in binding buffer, and then stained with Annexin V-APC and PI buffer for 30 minutes at room temperature in the dark. Then, the ratios of apoptotic cells were measured by flow cytometry (CytoFLEX LX, Beckman Coulter, CA).
Cell cycle analysis was done using the cell cycle kit (KGA512, KeyGEN BioTECH, Nanjing, China) following the manufacturer’s instructions. Hepatocellular carcinoma cells were collected and washed with PBS for 3 times. Then, cells were carefully resuspended and fixed with 70% ethanol overnight at 4°C. Cells were stained with PI/RNase buffer for 20 minutes at room temperature and shielded from light. Cell cycle of the stained HCC cells were then measured immediately by flow cytometry (CytoFLEX LX, Beckman Coulter, CA) and percentages of cells in different cell cycle phases were calculated.

Hu X., Liu T., Li L., Gan H., Wang T., Pang P, & Mao J. (2023). Fibulin-2 Facilitates Malignant Progression of Hepatocellular Carcinoma. The Turkish Journal of Gastroenterology, 34(6), 635-644.

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Immunophenotyping of Peripheral Blood Lymphocytes

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Flow cytometry was employed to examine the immunophenotype of lymphocytes found in peripheral blood. The process involved obtaining a whole blood sample, which was then treated with a set of monoclonal human antibodies comprising anti-CD4 BV421, anti-CD3 PerCp, anti-CD8 BV605, anti-CD19 FITC, anti-CD45 Alexa Fluor 700, anti-CD56 BV650, anti-CD16 BV650, anti-TLR-2 APC, anti-TLR-4 PE, anti-TLR-7 PE, anti-TLR-8 APC, anti-TLR-3 PE, and anti-TLR-9 APC (Biolegend, San Diego, CA, United States). Following the antibody staining, a lysing buffer (BD, Franklin Lakes, NJ, United States) was applied to eliminate red blood cells, and the resulting cells were thoroughly washed and evaluated using a CytoFLEX LX instrument (Beckman Coulter, Indianapolis, IN, United States). Subsequently, data analysis was carried out utilizing the Kaluza Analysis program (sample analysis in Figure 1—CLL and Figure 2—CVID) The CytoFLEX LX flow cytometer was internally quality controlled using CytoFLEX Ready to Use Daily QC Fluorosphere reagents (Beckman Coulter, Indianapolis, IN, United States).

Mertowska P., Smolak K., Mertowski S, & Grywalska E. (2023). Unraveling the Role of Toll-like Receptors in the Immunopathogenesis of Selected Primary and Secondary Immunodeficiencies. Cells, 12(16), 2055.

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Akata-EBV Infection Assay for Neutralization

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Akata-EBV, which is GFP positive, was prepared using the Akata cell line as previously described (27 (link), 29 (link), 46 (link)). Five thousand HNE1 cells were seeded in each well of 96-well plates. We allowed cells to grow overnight. We prepared a 2-fold serum dilution series with 10% FBS DMEM containing antibiotics. We added an equal amount of Akata EBV to each well. We allowed serum binding to virus for 1 h at 37°C with 5% CO₂. We discarded medium of HNE1 and replaced it with the serum-virus mix. We allowed virus infection for 2 h at 37°C with 5% CO₂. We replaced it with fresh 10% FBS DMEM containing antibiotics. Twenty-four hours later, the cells were digested with trypsin, and GFP signal was detected by flow cytometry (CytoFLEX LX; Beckman).
For B cell line neutralization evaluation, 10,000 Akata(−) cells in 50 μL FBS-containing medium were seeded in each well of 96-well plates. We prepared 100 μL serum-virus mix/well as previously described. We allowed continuous virus infection for 24 h. GFP signal was detected by flow cytometry (CytoFLEX LX; Beckman).

Kong X.W., Zhang X., Bu G.L., Xu H.Q., Kang Y.F., Sun C., Zhu Q.Y., Ma R.B., Liu Z., Zeng Y.X., Zeng M.S, & Hu Z.L. (2022). Vesicular Stomatitis Virus-Based Epstein-Barr Virus Vaccines Elicit Strong Protective Immune Responses. Journal of Virology, 96(9), e00336-22.

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Flow Cytometric Immunophenotyping of Blood Lymphocytes

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The analysis of lymphocyte immunophenotype in peripheral blood was performed through the use of flow cytometry, a precise and accurate approach to cell analysis. A whole blood sample was collected and treated with a set of monoclonal human anti-bodies consisting of anti-CD45 AF700, anti-CD3 PerCp, anti-CD4 BV421, anti-CD8 BV605, anti-CD19 FITC, anti-CD56 BV650, and anti-CD16 BV650, as well as anti-TLR2 APC, anti-TLR3 PE, anti-TLR4 PE, anti-TLR7 PE, anti-TLR8 APC, and anti-TLR9 APC antibodies (BioLegend, San Diego, CA 92121, USA). Subsequently, a lysing buffer was utilized to remove any red blood cells, and the remaining cells were thoroughly washed and assessed through the use of a CytoFLEX LX instrument, which is a sophisticated flow cytometer (Beckman Coulter, Indianapolis, IN, USA). The resulting data were analyzed using the Kaluza Analysis program, as demonstrated in Figure 2. The CytoFLEX LX flow cytometer was subjected to daily quality control using CytoFLEX Ready to Use Daily QC Fluorospheres reagents (Beckman Coulter, Indianapolis, IN, USA).

Smok-Kalwat J., Mertowska P., Mertowski S., Góźdź S., Korona-Głowniak I., Kwaśniewski W, & Grywalska E. (2024). Analysis of Selected Toll-like Receptors in the Pathogenesis and Advancement of Non-Small-Cell Lung Cancer. Journal of Clinical Medicine, 13(10), 2793.

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Quantification of Mitochondrial ROS Levels

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Staining of isolated mitochondrial mtROS levels was measured using CytoFLEX LX (Beckman Coulter)³⁸ . Mitochondrial assay solution [MAS, 3 × ; 210 mmol/L sucrose, 660 mmol/L mannitol, 30 mmol/L KH₂PO₄, 15 mmol/L MgCl₂, 6 mmol/L HEPES, 3 mmol/L EGTA, and 0.6% (w/v) fatty acid-free BSA, pH 7.2] was used for staining. Rotenone (2 μmol/L), malonate (10 mmol/L), and antimycin A (4 μmol/L) were used as inhibitors of mitochondrial complexes I, II, and III, respectively. Mitochondria were isolated by the method described earlier. Succinate (10 mmol/L) and mitochondrial inhibitors were incubated with MitoSOX (1 μmol/L) and mitochondria in MAS (diluted to 1 ×, pH 7.2) at room temperature (20 °C) for 1 h. Afterward, mitochondria were washed by MAS with substrate and inhibitors to clear the surplus dye, resuspended in MAS with substrate and inhibitors, and measured using CytoFLEX LX (Beckman Coulter).
mtROS level of MRC-5 cells was measured using Synergy H1 Hybrid Multi-Mode Reader (Bio-Tek Instruments). Cells were incubated with or without chlorpropamide for 1, 3, 6, 9, 12, and 24 h. These cells were then washed by PBS, stained by MitoSOX (5 μmol/L) for 30 min, washed by PBS again, and measured using a Synergy H1 Hybrid Multi-Mode Reader (Bio-Tek Instruments). 12 wells were measured for each group and MitoTracker® Green FM (200 nmol/L) was used for normalization.

Mao Z., Liu W., Huang Y., Sun T., Bao K., Feng J., Moskalev A., Hu Z, & Li J. (2021). Anti-aging effects of chlorpropamide depend on mitochondrial complex-II and the production of mitochondrial reactive oxygen species. Acta Pharmaceutica Sinica. B, 12(2), 665-677.

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Acoustic Cavitation-Mediated Gene Transfection

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The pCMV- IRES-EGFP (pEGFP) plasmid was used to create the pEGFP-GVs complexes, which were then incubated with HEK293 cells for 6 h (pEGFP-GVs@HEK293 cells). The resulting pEGFP-GVs@HEK293 cells were then subjected to 1 min of acoustic irradiation at 0, 0.1, 0.2, or 0.5 MPa pressure, followed by observation under a fluorescence microscope (EVOS M7000, Thermo Fisher, USA) after 48 h or quantitative analysis of their transfection efficiencies by flow cytometry (CytoFLEX LX, Beckman, USA). To compare the gene transfection efficiencies mediated by intracellular cavitation and extracellular cavitation, pEGFP-GVs complexes were incubated with HEK293 cells, B1610 cells, MC38 cells, and C6 cells for 6 h, yielding pEGFP-GVs@HEK293 cells, pEGFP-GVs@B1610 cells, pEGFP-GVs@MC38 cells and pEGFP-GVs@C6 cells, respectively. For 1 min, these pEGFP-GVs@cells were irradiated at 0.5 MPa of acoustic pressure. In terms of extracellular cavitation, equivalent pEGFP-GVs complexes were added to these HEK HEK293 cells, B1610 cells, MC38 cells, and C6 cells and immediately treated with acoustic irradiation at 0.5 MPa for 1 min. These irradiated cells were examined under a fluorescence microscope (EVOS M7000, Thermo Fisher, USA) after 48 h and quantitatively analyzed using flow cytometry (CytoFLEX LX, Beckman, USA).

Xie L., Wang J., Song L., Jiang T, & Yan F. (2023). Cell-cycle dependent nuclear gene delivery enhances the effects of E-cadherin against tumor invasion and metastasis. Signal Transduction and Targeted Therapy, 8, 182.

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Mesangial Cell Proliferation Assays

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Cell proliferation was assessed using Cell Counting Kit-8 (CCK-8) (Dojindo, Japan) and a 5-ethynyl-2-deoxyuridine (EdU) labeling/detection kit (BeyoClick™ EdU Cell Proliferation Kit with Alexa Fluor 555, Beyotime, China). For CCK-8 assay, mesangial cells were seeded in a 96-well plate exposed to high glucose (30 mM) with or without pretreatment of different concentrations of SA for 24 h. Next, 10 µl CCK-8 reagent was added to each well containing 100 µl DMEM medium and the plate was incubated for 2 h at 37°C. Finally, a microplate reader (BioTek, United States) was used to detect the absorbance at 450 nm.
For EdU assay, cells were cultured in a 12-well plate and incubated with EdU for 2 h. Trypsin was added into the microcentrifuge tube to digest the cells, which were then fixed with 4% paraformaldehyde for 15 min and permeated with 0.3% Triton X-100 for another 15 min. Next, cells were incubated with the Click Reaction Mixture for 30 min at room temperature in a dark place following the manufacturer’s instructions. Finally, cells were re-suspended in phosphate buffered saline (PBS) for flow cytometric analysis on the Beckman Coulter CytoFLEX LX (Beckman Coulter, United States). Data were analyzed using FlowJo software version 10 (Treestar).

Wang Z., Chen Z., Wang X., Hu Y., Kong J., Lai J., Li T., Hu B., Zhang Y., Zheng X., Liu X., Wang S., Ye S., Zhou Q, & Zheng C. (2022). Sappanone a prevents diabetic kidney disease by inhibiting kidney inflammation and fibrosis via the NF-κB signaling pathway. Frontiers in Pharmacology, 13, 953004.

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Isolation and Characterization of Murine Corneal Cells

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Dissected mouse corneal stroma and epithelium were separated by treatment with dispase (1 hour at RT in supplemental hormonal epithelial medium [SHEM] containing 50 mg/mL dispase-II and 100 mM sorbitol). Single-cell suspensions of mouse corneal stromal cells were obtained by incubating dissected stroma in collagenase for 45 minutes and filtering through a 35-µm cell strainer cap. The filtered cells were stained for viability and fluorescent antibodies that detected CD11b (integrin αM, expressed by many leukocytes, including monocytes, neutrophils, natural killer cells, granulocytes, and macrophages), CD11c (integrin αX, a marker of dendritic cells), Ly6G (lymphocyte antigen 6 family member G, a marker for monocytes, granulocytes, and neutrophils) (all from BioLegend, San Diego, CA, USA), F4/80 (a marker of murine macrophages), Ly6C (lymphocyte antigen 6 family member C, a marker for monocytes, macrophages, and endothelial cells) (BD Pharmingen, Franklin Lakes, NJ, USA); washed with PBS with 2% fetal bovine serum; fixed in 2% paraformaldehyde; filtered through a mesh; and analyzed using Beckman Coulter Cytoflex LX (Beckman Coulter, Brea, CA, USA) as earlier.¹⁵ (link)

Swamynathan S., Campbell G., Sohnen P., Kaur S., St. Leger A.J, & Swamynathan S.K. (2024). The Secreted Ly6/uPAR-Related Protein 1 (Slurp1) Modulates Corneal Angiogenic Inflammation Via NF-κB Signaling. Investigative Ophthalmology & Visual Science, 65(1), 37.

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Isolation and Analysis of Hematopoietic Stem Cells

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For flow cytometry analysis and isolation of hematopoietic stem and progenitor cells, cells were stained with relevant antibodies^{51 (link)} in PBS with 2% fetal bovine serum for 30−45 min on ice. Antibody clones that were used: Sca-1-PE-Cy7, c-Kit-APC, CD150-PE, CD48-BV421, CD45.1-FITC, CD45.2 PE-Cy5, Gr-1-PE-Cy5, Mac1-PE-Cy5, IgM-PE-Cy5, CD3-PE-Cy5, CD4- PE-Cy5, CD8-PE-Cy5, CD45R-PE-Cy5 and Ter-119-PE-Cy5. Detailed antibody information is summarized in Supplementary Table S6. HSPCs were stained with a lineage antibody cocktail (Gr-1, Mac1, CD3, CD4, CD8, CD45R, TER119 and B220), Sca-1, c-Kit, CD150 and CD48. Cell types were defined as followed: LSK compartment (Lin⁻Sca-1⁺c-Kit⁺), LT-HSC (LSK CD150⁺CD48⁻), ST-HSC (LSK CD150⁻CD48⁻), MPP2 (LSK CD150⁺CD48⁺) and MPP3/4 (LSK CD150⁻CD48⁺). B cells (CD45.2⁺Mac1⁻Gr-1⁺B220⁺), T cells (CD45.2⁺Mac1⁻Gr-1⁺CD3⁺) and myeloid cells (CD45.2⁺Mac1⁺Gr-1⁻). Samples were analyzed on a flow cytometer (CytoFLEX LX, Beckman). For sorting HSCs, lineage antibody cocktail-conjugated paramagnetic microbeads and MACS separation columns (Miltenyi Biotec) were used to enrich Lin⁻ cells before sorting. Stained cells were re-suspended in PBS with 2% FBS and sorted directly using the Beckman moflo Astrios EQ (Beckman). Flow cytometry data were analyzed by FlowJo (BD) software.

Jiang L., Ye Y., Han Y., Wang Q., Lu H., Li J., Qian W., Zeng X., Zhang Z., Zhao Y., Shi J., Luo Y., Qiu Y., Sun J., Sheng J., Huang H, & Qian P. (2024). Microplastics dampen the self-renewal of hematopoietic stem cells by disrupting the gut microbiota-hypoxanthine-Wnt axis. Cell Discovery, 10, 35.

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Mitochondrial Membrane Potential Assay

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To detect mitochondrial membrane potential, cardiomyocytes were incubated with JC-1 (Beyotime, Shanghai, China) for 20 min at 37 °C. Cardiomyocytes were analyzed within 1 h using flow cytometry (CytoFLEX LX, Beckman, USA). All data were analyzed using EXPO32 ADC analysis software (Beckman, Washington, USA). For flow cytometry analysis, unstained cells were used as controls to identify the forward scatter/side scatter (FSC/SSC) gates used in the same experiments. A total of 10,000 cells was detected in each sample [30 (link)].

Jiang M.Y., Man W.R., Zhang X.B., Zhang X.H., Duan Y., Lin J., Zhang Y., Cao Y., Wu D.X., Shu X.F., Xin L., Wang H., Zhang X., Li C.Y., Gu X.M., Zhang X, & Sun D.D. (2023). Adipsin inhibits Irak2 mitochondrial translocation and improves fatty acid β-oxidation to alleviate diabetic cardiomyopathy. Military Medical Research, 10, 63.

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