Facscalibur cytometer

Manufactured by BD

Sourced in United States, Germany, France, Italy, Spain, United Kingdom, China

The FACSCalibur cytometer is a flow cytometry instrument designed for multiparameter analysis of cell populations. It utilizes a dual-laser excitation system to detect and measure various cellular characteristics, including size, granularity, and fluorescence intensity.

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Lab products found in correlation

Cellquest software, by BD (117 mentions) Cellquest pro software, by BD (61 mentions) Propidium iodide, by Merck Group (39 mentions) Cellquest pro, by BD (37 mentions) Rnase a, by Merck Group (14 mentions) Cellquest, by BD (11 mentions) Annexin 5 fitc, by BD (10 mentions) Cytofix cytoperm, by BD (9 mentions) Cytofix cytoperm kit, by BD (9 mentions) Fitc annexin 5 apoptosis detection kit, by BD (8 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions) Cellquest pro 5, by BD (8 mentions) Mca2226f, by Bio-Rad (8 mentions) Annexin 5 fitc apoptosis detection kit, by Beyotime (7 mentions) Facscanto 2, by BD (7 mentions) Trypsin edta, by Thermo Fisher Scientific (7 mentions) Facsdiva software, by BD (6 mentions) Bovine serum albumin (bsa), by Merck Group (6 mentions) Propidium iodide, by Thermo Fisher Scientific (6 mentions) Cell quest pro version 6, by BD (6 mentions)

Close spelling variants of this product

LSR-II or FACSCalibur flow cytometers Vantage BD FACSCalibur flow cytometer FACSCalibur Flow Cytometer system FACScalibur Becton Dickinson cytometer BD FACSCalibur fluorescent-activated flow cytometer FACSCalibur benchtop cytometer Single-laser FACScalibur cytometer FACSCalibur™ fow cytometer FACSCalibur dual laser flow-cytometer Becton Dickinson FACSCalibur flow cytometer

1 030 protocols using facscalibur cytometer

Quantifying MSC Differentiation and Phenotype

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Flow cytometry was used to examine the coating efficiency VCAM 1 antibody. We stained 10⁶ V-MSCs and MSCs with 5 μL ALEXA-labeled anti-mouse CD106 (Becton Dickinson, San Jose, CA, USA) at room temperature for 20 min. Unstained cells were used as a control. The cells were then washed twice, resuspended in 500 μL PBS, and examined using a BD FACS Calibur cytometer (Becton Dickinson).
To examine the differentiation and phenotypes of MSCs, the MSCs were seeded in the wells of 24-well plates at a concentration of 5×10⁴ cells/well. Differentiation of MSCs into osteogenic and adipogenic lineages was assayed as reported previously [10 (link)]. Briefly, MSCs were harvested, digested with trypsin-EDTA, and stained with ALEXA-labeled anti-mouse CD105, phycoerythrin (PE)-conjugated anti-mouse CD90, fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD45, and PE Cy-conjugated anti-mouse CD11b (Becton Dickinson) and analyzed on a BD FACS Calibur cytometer.

Chen Q., Li Y., Chen Z., Du H, & Wan J. (2019). Anti-VCAM 1 Antibody-Coated Mesenchymal Stromal Cells Attenuate Experimental Colitis via Immunomodulation. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 4457-4468.

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Apoptosis and Cell Cycle Analysis of CAP-Treated Glioblastoma Cells

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After treating U-87 MG cells with CAPs IC50 doses obtained from the MTT assay, the cells were incubated for 48 h and 72 h in the helium and argon gas plasma groups, respectively. The Annexin V-FITC/PI apoptosis detection kit (Bender MedSystems, Austria) was utilized to quantify early and late apoptosis of U-87 MG cells after CAP treatment, following the manufacturer's instructions. In brief, after treatment, cells were washed in PBS, then resuspended in 100 μl of binding buffer and stained with 5 μl FITC-conjugated Annexin-V for 15 min in darkness at room temperature. Subsequently, samples were washed, resuspended in 250 μl binding buffer, and incubated with 5 μl Propidium Iodide (PI) from Sigma-Aldrich, MO, USA for 10 min. The results were analyzed using the BD FACSCalibur cytometer. Additionally, the DNA content was determined to quantify the G1, S, and G2/M phases of the cell cycle. For this purpose, following treatment, the cells were detached using 0.25% trypsin–EDTA (GIBCO, NY, USA). After detachment, the cells underwent PBS washes, followed by fixation with ice-cold 70% ethanol for 2 h. Subsequently, the cells were washed using PBS, treated with 50 μg/ml RNase A (Bio Basic, Canada) for 30 min, and then incubated with Propidium Iodide (PI; Sigma-Aldrich, USA). Flow cytometry analysis was carried out using the BD FACSCalibur cytometer.

Bakhtiyari-Ramezani M., Nohekhan M., Akbari M.E., Abbasvandi F., Bayat M., Akbari A, & Nasiri M. (2024). Comparative assessment of direct and indirect cold atmospheric plasma effects, based on helium and argon, on human glioblastoma: an in vitro and in vivo study. Scientific Reports, 14, 3578.

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Exendin-4 Modulates Mesenchymal Stem Cell Characteristics

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The CXCR4 expression, apoptotic rate and cell cycle distribution of MSC were evaluated using flow cytometry. After treatment with different concentrations of Ex-4 (0–20 nm/L) for 24 h, MSC at P₃ were used to detect CXCR4 expression. After being washed in PBS, each group of cells was stained with FITC-labeled CXCR4 (Bioss, China) or FITC-labeled IgG for 30 min at room temperature.
The number of apoptotic cells was analyzed quantitatively using the Annexin V–FITC/PI Apoptosis Detection Kit (BD Biosciences, USA). After treatment, the cells were harvested, resuspended in 200 μl of binding buffer, and then incubated with 5 μl of Annexin V–FITC/binding buffer mixture (30 min, 37 °C) in the dark. Subsequently, the cells were incubated with 10 μl of propidium iodide for 5 min and immediately analyzed by bivariate flow cytometry using a BD FACSCalibur cytometer.
For the cell cycle analysis, MSC were treated with Ex-4 (0–20 nm/L) for 24 h. The cells were then resuspended in PBS and fixed with ice-cold 70% ethanol for 24 h. The fixed cells were rinsed and resuspended in 50 μg/ml propidium iodide for 30 min in the dark. Flow cytometric analyses were performed using a BD FACSCalibur cytometer.

Zhou H., Li D., Shi C., Xin T., Yang J., Zhou Y., Hu S., Tian F., Wang J, & Chen Y. (2015). Effects of Exendin-4 on bone marrow mesenchymal stem cell proliferation, migration and apoptosis in vitro. Scientific Reports, 5, 12898.

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Measuring Homologous Recombination Capacity

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To measure HR capacity, PDCs were co-transfected with pDR-GFP + pCBASceI (a gift from Dr. Maria Jasin, Addgene #26475 and #26477) or pDR-GFP + pFUGW-RFP (transfection efficiency control) (1:1) for a total of 2.5 μg DNA using lipofectamine LTX with Plus Reagent (Invitrogen) according to the manufacturer’s protocol in OptiMEM (Gibco) in 6-well plates (Corning 3506). Six hours post-transfection, media was replaced with media containing vehicle or inhibitors. Cells were incubated for an additional 48 h, collected, and analyzed for GFP and RFP expression using a FACSCalibur cytometer with CellQuest Pro software (BD Biosciences).
To evaluate the effects of DDR inhibitors on HR capacity, PDCs were transfected with pDR-GFP and selected with 1–10 μg/ml puromycin 48 h later. Post-selection, PDR-GFP stable lines were seeded in 6-well plates, allowed to attach overnight, and transfected with pCBASceI or pFUGW-RFP using lipofectamine LTX as above. Six hours post-transfection, media was replaced with media containing vehicle or inhibitors. Cells were incubated for an additional 72 h, collected and analyzed for GFP expression using a FACSCalibur cytometer with CellQuest Pro software (BD Biosciences).

El Touny L.H., Hose C., Connelly J., Harris E., Monks A., Dull A.B., Wilsker D.F., Hollingshead M.G., Gottholm-Ahalt M., Alcoser S.Y., Mullendore M.E., Parchment R.E., Doroshow J.H., Teicher B.A, & Rapisarda A. (2021). ATR inhibition reverses the resistance of homologous recombination deficient MGMTlow/MMRproficient cancer cells to temozolomide. Oncotarget, 12(21), 2114-2130.

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Cell Apoptosis and Cell Cycle Analysis

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50 000 cells of each line were cultured in a 6-wells plates with onvansertib (Selleckchem, NMS-P937, NMS1296937) and/or cisplatin for 48 h.
Apoptotic and dead cells were stained with Annexin V-APC (Biolegend, 8 μg/ml) and propidium iodide (PI, Biolegend, 0.5 mg/ml) for 15 min at 4 °C and analyzed with FACSCalibur cytometer (BD Biosciences).
To analyze cell cycle and polyploidy, 200 000 cells were cultured with onvansertib or cisplatin for 24 h and conserved in ethanol 70% at -20 °C. They were incubated in PBS containing 3 μg/ml RNase A and 40 μg/ml PI for 30 min at 4 °C and analyzed with FACSCalibur cytometer (BD Biosciences).

Hagege A., Ambrosetti D., Boyer J., Bozec A., Doyen J., Chamorey E., He X., Bourget I., Rousset J., Saada E., Rastoin O., Parola J., Luciano F., Cao Y., Pagès G, & Dufies M. (2021). The Polo-like kinase 1 inhibitor onvansertib represents a relevant treatment for head and neck squamous cell carcinoma resistant to cisplatin and radiotherapy. Theranostics, 11(19), 9571-9586.

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Apoptosis and Mitochondrial Potential Assays

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2 × 10 4 per well cells were seeded in 6-well plates and left for 24-hour growth in 5% CO2 incubator at 37°C. The cells were then exposed to the drug for indicated times and concentrations. At the end of the incubation period, the culture medium was removed and the cells were washed twice with cold phosphate-buffered saline (PBS; Gibco, 10010023) and harvested through trypsinization. FACS Calibur cytometer (BD, San Jose, USA) was used to analyze the apoptosis rate using a FITC Annexin V Apoptosis Detection Kit I (MultiSciences (Lianke) Biotech, AP101) according to the manufacturer's instructions.
Mitochondrial membrane potential detection.
Cells were treated and harvested as mentioned above, and then resuspended in 1mL PBS and moved to 1.5 mL centrifuge tube. 10 μg/mL JC-1 (Sigma-Aldrich, T4069) was added and the cells were incubated for 30 min at 37°C without light. After the incubation period, the cells were washed twice with PBS to remove the unbinding JC-1 and then resuspended in 500 μL PBS. FACS Calibur cytometer (BD, San Jose, USA) was used to analyze the change of mitochondrial membrane potential.

Jiang L.Y., Zeng Y., Ai L., Yan H., Yang X., Luo P.H., Yang B., Xu Z., & He Q. (2022). Reduced HMGB1 expression contributed to lapatinib-induced cutaneous injury.

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Apoptosis and Cell Cycle Analysis in Cultured Cells

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Cells (5×10⁵ cells/well) were grown in 6-well plates for 72 h. Apoptosis was measured using the Caspase-3 active form (cleaved Caspase-3) mAb Apoptosis Kit, FITC (BD bioscience). The signals were detected by a FACScalibur cytometer (BD biosciences). The percentage of apoptotic cells was determined using FlowJo software (Tree Star).
Cell cycle analysis was performed using APC BrdU flow kit (BD bioscience). The signals were detected a by FACScalibur cytometer (BD biosciences) and analyzed using FlowJo software (Tree Star).

Garancher A., Lin C.Y., Morabito M., Richer W., Rocques N., Larcher M., Bihannic L., Smith K., Miquel C., Leboucher S., Herath N.I., Dupuy F., Varlet P., Haberler C., Walczak C., El Tayara N., Volk A., Puget S., Doz F., Delattre O., Druillennec S., Ayrault O., Wechsler-Reya R.J., Eychène A., Bourdeaut F., Northcott P.A, & Pouponnot C. (2018). NRL and CRX define photoreceptor identity and reveal subgroup-specific dependencies in medulloblastoma. Cancer cell, 33(3), 435-449.e6.

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Apoptosis Quantification by Flow Cytometry

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Cells (approximately 1 × 10⁶ cells per well) were collected and centrifuged at 1,000 × g for 5 min at room temperature, resuspended in ice-cold PBS, centrifuged at 1,000 × g for 5 min, and washed. The cells were resuspended by adding 500 μL of 1 × binding buffer. Subsequently, 5 μL annexin V–fluorescein isothiocyanate (FITC) staining solution and 5 μL PI staining solution were added to the suspension, mixed well, and incubated for 30 min at room temperature. Fluorescence intensity was measured using a BD FACSCalibur cytometer (Becton–Dickinson), and the apoptotic rates of cells were analyzed using the FlowJo software (Tree Star). The cells in Q1 (left upper quadrant), Q2 (right upper quadrant), Q3 (right lower quadrant), and Q4 (left lower quadrant) represent dead cells, late apoptotic cells, early apoptotic cells, and living cells, respectively. Three independent repeats of experiments were performed.

Shi W., Zhang S., Ma D., Yan D., Zhang G., Cao Y., Wang Z., Wu J, & Jiang C. (2020). Targeting SphK2 Reverses Acquired Resistance of Regorafenib in Hepatocellular Carcinoma. Frontiers in Oncology, 10, 694.

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Phagocyte Oxidative Activity Quantification

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Quantification of the phagocytes’ oxidative activity (monocytes and neutrophils) was performed by flow cytometry from heparinized whole blood using BURSTTEST kit, Phagoburst, (IVD) (BD Pharmingen). The kit contains unlabeled opsonized E. coli bacteria, PMA and fMLP as particulate, high and low physiological stimuli. It quantifies the percentage of leukocytes (monocytes and neutrophils) which oxidize the fluorogenic substrate dihydrorhodamine (DHR123) to rhodamine 123, and their enzymatic activity (amount of rhodamine 123). The test was performed using BD FACSCalibur cytometer (Becton Dickinson) and CellQuest software (BD); daily check-up of cytometer performances was performed using BD Calibrite 3 Beads kit (BD Biosciences) and BD FACSComp software (BD). The percentage of cells that have produced ROS (% gated) and their oxidative activity (mean fluorescence, GeoMean) were analyzed. When presented as stimulation index, the results were compared to the given stimuli (fMLP, E. coli and PMA) based on unstimulated control samples (ratio between the values obtained for stimulated and unstimulated samples). Test was performed according to the manufacturer and published studies.17 (link)

Munteanu A.N., Surcel M., Isvoranu G., Constantin C, & Neagu M. (2022). Healthy Ageing Reflected in Innate and Adaptive Immune Parameters. Clinical Interventions in Aging, 17, 1513-1526.

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Cell Cycle Analysis of Inducible Protein Expression

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Flp-In T-REx-293-ZC3H12B and Flp-In T-REx-293-ZC3H12B-D196A were treated with 1μg/ml doxycycline for induction of recombinant protein expression for 72 h. Then, the cells were trypsynized, washed with PBS two times, resuspended in 2 mL of PBS and fixed by addition of 6 mL of ice-cold 96% ethanol. After fixation the cells were washed two times with PBS and stained with PI/SAP solute ion (0.05 mg/mL propidium iodide, 1 mg/mL RNase A, 0.02% saponin, 0.0001% Triton-X-100 in PBS) by incubation for 15 min at 37°C. Immediately after staining, the cells (200,000 of FSC/SSC gated cells/sample) were analyzed for propidium iodide fluorescence intensity by flow cytometry using the BD FACSCalibur cytometer (Becton Dickinson). BD CellQuest Pro software (Becton Dickinson) was used for data acquisition. The percentage of cells in each cycle was calculated with the FlowJo (v10.4.2, FlowJo) Cell Cycle analysis platform using the Watson (pragmatic) model.

Wawro M., Wawro K., Kochan J., Solecka A., Sowinska W., Lichawska-Cieslar A., Jura J, & Kasza A. (2019). ZC3H12B/MCPIP2, a new active member of the ZC3H12 family. RNA, 25(7), 840-856.

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