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Facsort

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The FACSort is a flow cytometry instrument designed for the analysis and sorting of cells. It is capable of detecting and measuring various characteristics of individual cells, such as size, granularity, and the presence of specific proteins or markers. The FACSort provides researchers with a tool to isolate and purify specific cell populations from complex samples for further study or applications.

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Lab products found in correlation

Cellquest software, by BD (16 mentions) Modfit software, by Verity Software House (7 mentions) Propidium iodide, by Merck Group (3 mentions) Fitc annexin 5 apoptosis detection kit, by BD (3 mentions) Cellquest, by BD (3 mentions) Rnase a, by Merck Group (3 mentions) Cellquest 3, by BD (3 mentions) Fluorescent activated cell sorter facscount system, by BD (2 mentions) Yoyo 1, by Thermo Fisher Scientific (2 mentions) Ams 1 loader, by Cytek (2 mentions) Cellquest pro software, by BD (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Annexin 5 fitc apoptosis detection kit, by Thermo Fisher Scientific (2 mentions) Cellquest software version 3, by BD (2 mentions) Annexin 5 fitc pi apoptosis detection kit, by Beyotime (2 mentions) Propidium iodide, by Keygen Biotech (2 mentions) Facscanto 2, by BD (2 mentions) Cellquest pro, by BD (2 mentions) Annexin 5 fitc pi kit, by Merck Group (1 mentions) Fortessa x20 flow cytometer, by BD (1 mentions)

Close spelling variants of this product

FACSort flow cytometer (169 citations) FACSort cytometer (13 citations) FACSort system (5 citations) FACSort instrument (5 citations) FACStar Plus dual laser FACSort system (4 citations) FACSort Cellquect software (3 citations) Becton-Dickinson FACSort flow cytometer (2 citations) FACSort machine (2 citations) FACSort CellQuest (2 citations) FACSort device (2 citations)

172 protocols using facsort

1

Osteosarcoma Cell Surface Marker and miR-335 Expression Analysis

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For analysis of cell surface markers, osteosarcoma cells were harvested and resuspended in PBS/0.5% normal rabbit serum (Sigma-Aldrich), and blocked on ice for 15 min. Cells were subsequently labelled with Alexa Fluor® 647 anti-human Stro-1 antibody (BioLegend) and PE anti-human CD117 (c-kit) antibody (BioLegend) for 60 min and maintained on ice until analysis. The expression was assessed by flow cytometry and data were analyzed. The double positive (DP) and double negative (DN) cells were then sorted and collected using a Becton–Dickinson FACSort (San Jose).
In order to divide the cells into different subpopulations according to their miR-335 expression status, we incubated the cells with miR-335-5p Hu-Cy5 SmartFlare™ RNA Detection Probe (Millipore), overnight for 16 h. The fluorescence was then detected and cells were sorted into miR-335-high and miR-335-low subpopulation using a Becton–Dickinson FACSort (San Jose). The collected cells were then send for subsequent experiments.
Guo X., Yu L., Zhang Z., Dai G., Gao T, & Guo W. (2017). miR-335 negatively regulates osteosarcoma stem cell-like properties by targeting POU5F1. Cancer Cell International, 17, 29.
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2

Cell Cycle Analysis by Flow Cytometry

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Cells were harvested (48 h post-transfection) in flow cytometry tubes and centrifuged at 1,000× g for 5 min to pellet the cells. The pellets were washed with PBS and fixed with 70% cold ethanol at 4°C overnight. After washing again with PBS, the cells were incubated with RNAaseA (0.1 mg/mL) for 30 min, followed by incubation with propidium iodide (50 µg/mL) for 30 min at room temperature. Cell cycle analysis was performed using a flow cytometer (FACSort; Becton Dickinson, Franklin Lakes, NJ, USA). The percentage of cells at G0/G1, S, and G2/M phases was analyzed using the FlowJo software 3.1 (TreeStar, Inc., Ashland, OR, USA).
Wang R., Ma Q., Ji L., Yao Y., Ma M, & Wen Q. (2018). miR-622 suppresses tumor formation by directly targeting VEGFA in papillary thyroid carcinoma. OncoTargets and therapy, 11, 1501-1509.
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3

Annexin V-FITC/PI Cell Death Assay

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Cell death, including apoptosis and necrosis, was assessed by staining with an annexin V–FITC/PI kit (Sigma), according to the manufacturer’s instructions. Briefly, the cells were cultured with various concentrations of bicyclol for 48 h, and then 1 × 106 cells were harvested and washed twice with ice-cold PBS. The apoptotic (Annexin V+/PI-) or necrotic cells (Annexin V+/PI+) were evaluated by double staining with annexin V–FITC and PI in binding buffer using flow cytometry (FAC sort, Becton Dickinson).
Wang Y., Nie H., Zhao X., Qin Y, & Gong X. (2016). Bicyclol induces cell cycle arrest and autophagy in HepG2 human hepatocellular carcinoma cells through the PI3K/AKT and Ras/Raf/MEK/ERK pathways. BMC Cancer, 16, 742.
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4

Mitochondrial Dysfunction Assay with Zey

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Cells were incubated in the presence of different concentrations of Zey (HeLa: 0, 3.27, 6.54 and 13.08 µM; CaSki: 0, 1.64, 3.27 and 6.54 µM) for 24 h and were then stained for 20 min with 10 µg/mL JC-1, which accumulates in mitochondria in a potential-dependent manner. The cells were collected, washed with PBS, and analyzed using a FACSort flow cytometer (Becton-Dickinson, CA, USA). Production of green fluorescence and reduction of the red fluorescence indicates loss of mitochondrial transmembrane potential.
Zhang L., Huo X., Liao Y., Yang F., Gao L, & Cao L. (2017). Zeylenone, a naturally occurring cyclohexene oxide, inhibits proliferation and induces apoptosis in cervical carcinoma cells via PI3K/AKT/mTOR and MAPK/ERK pathways. Scientific Reports, 7, 1669.
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5

Apoptosis Induction by AMRI-59 and IR

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Cells were seeded at a density of 1 × 105 cells and incubated with or without 10 or 30 nM AMRI-59. After 16 h incubation, cells were exposed to 3 Gy IR and incubated for 48 h. Treated cells were harvested via trypsinization, washed twice with cold PBS, and resuspended in 300 μL of a 5 μg/mL propidium iodide (PI, Sigma-Aldrich) solution. The apoptotic fraction was evaluated using a FACSort flow cytometer (Becton Dickinson).
Hong W.G., Kim J.Y., Cho J.H., Hwang S.G., Song J.Y., Lee E., Chang T.S., Um H.D, & Park J.K. (2017). AMRI-59 functions as a radiosensitizer via peroxiredoxin I-targeted ROS accumulation and apoptotic cell death induction. Oncotarget, 8(69), 114050-114064.
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6

Flow Cytometric Analysis of Liposomes

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Cell suspensions were analyzed with a FORTESSA X20 flow cytometer (Beckton Dickinson, Franklin Lakes, NJ, USA). The extracellular fluorescence of the NBD-PE lipid present in the liposomes or the FITC conjugated to the nucleic acid was quenched using trypan blue as in [23 (link),44 (link),45 (link)]. The cell-associated fluorescence was then measured with a flow cytometer (FACSort; Becton Dickinson, Franklin Lakes, New Jersey) with λex = 488 nm; λem = 530 ± 30 nm. The fluorescence intensity was expressed as the mean fluorescence intensity of 10,000 events.
Perche F., Le Gall T., Montier T., Pichon C, & Malinge J.M. (2019). Cardiolipin-Based Lipopolyplex Platform for the Delivery of Diverse Nucleic Acids into Gram-Negative Bacteria. Pharmaceuticals, 12(2), 81.
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7

Stem Cell Characterization by Flow Cytometry

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For TRA-1-81 analysis, each 100 μl of cell suspension (106 cells) was used. Cells were incubated with the primary antibody mouse anti-TRA-1-81 (Santa Cruz) and then the secondary Alexa Fluor 568-labelled Goat anti-Mouse IgM (Invitrogen). For CD133 analysis, each 100 μl of cell suspension (106 cells) was used. Cells were incubated with the primary antibody CD133/1(AC133)-PE (Miltenyi Biotec). For cell cycle analysis, cells were processed with the Click-iT EdU Flow Cytometry Assay Kits (Invitrogen) according to the manufacturer's instructions. Briefly, the cells were harvested after incubation with EdU (10 μM) in medium. 100 μl of cell suspension (106 cells) for each sample were used. For the detection of cell apoptosis, each sample (106 cells) was subjected to Annexin V-PE staining20 (link). Cells were examined by fluorescence-activated cell sorting (FACS) using a flow cytometer (FACSort; Becton, Dickinson and Company), and the cell cycle populations were determined using ModFit software (Verity Software House, Inc.).
Duan S., Yuan G., Liu X., Ren R., Li J., Zhang W., Wu J., Xu X., Fu L., Li Y., Yang J., Zhang W., Bai R., Yi F., Suzuki K., Gao H., Esteban C.R., Zhang C., Belmonte J.C., Chen Z., Wang X., Jiang T., Qu J., Tang F, & Liu G.H. (2015). PTEN deficiency reprogrammes human neural stem cells towards a glioblastoma stem cell-like phenotype. Nature Communications, 6, 10068.
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8

Antibody-Mediated Dengue Virus Neutralization Assay

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Antibodies (1 mg/mL) were serially diluted in ten-fold steps and mixed with virus suspension at the multiplicity of infection of 0.01 for one hour at 37°C before adding to 2 × 105 U-937 cells/well in a 24-well cell culture plate. After virus adsorption, U-937 cells were incubated in a 5% CO2 incubator at 37°C for four days without removing the virus-antibody complexes. The presence of antibodies during incubation increased the sensitivity of the assay with minimal effect on the background level. Clarified culture media were collected and extracellular viruses were titrated in focus forming units (FFU) using the focus immunoassay titration method (Keelapang et al., 2004 (link)). For the enumeration of cell-associated viruses, infected cells were lysed by suspending cells in RPMI supplemented with 20% FBS, followed by three freeze-thaw cycles. Infectious viruses in clarified lysates were then titrated using PS cells.
Dengue virus-infected U-937 cells were enumerated by washing cells with PBS, fixing with 4% paraformaldehyde-PBS for one hour, permeabilizing cells with 0.1% Triton X-100-PBS, and then reacting sequentially with 4G2 and FITC-conjugated rabbit anti-mouse F(ab´)2 antibodies. For each sample, 20,000 events were acquired by using a fluorescence-activated cell sorter (FACSort, Becton Dickinson).
Keelapang P., Supasa P., Sriburi R., Puttikhunt C., Cardosa J., Kasinrerk W., Malasit P, & Sittisombut N. (2022). A group of infection-enhancing and focus size-reducing monoclonal antibodies recognized an ‘a and c’ strands epitope in the pr domain of Dengue Virus prM. Virus Research, 323, 199015.
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9

Almonertinib Effect on Calcein Accumulation

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The effect of almonertinib on the intracellular accumulation of fluorescent calcein (485 nm excitation and 535 nm emission), a known substrate of ABCB1 [45 (link)], was determined according to the method described by Gribar et al. [46 (link)]. In brief, cells were first trypsinized and resuspended in IMDM containing 5% FCS before calcein was added to 3 × 105 cells in 4 mL of IMDM in the presence of DMSO (control), 20 μM of almonertinib or tariquidar. The relative fluorescence intensity was detected and analyzed using a FACSort flow cytometer equipped with the CellQuest software (Becton-Dickinson) and the FlowJo software (Tree Star, Inc., Ashland, OR, USA), as described previously [47 (link), 48 (link)].
Wu C.P., Hung T.H., Lusvarghi S., Chu Y.H., Hsiao S.H., Huang Y.H., Chang Y.T, & Ambudkar S.V. (2021). The third-generation EGFR inhibitor almonertinib (HS-10296) resensitizes ABCB1-overexpressing multidrug-resistant cancer cells to chemotherapeutic drugs. Biochemical pharmacology, 188, 114516.
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10

Apoptosis and Cell Surface Marker Analysis

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Cells were fixed in 70% ethanol and stained in PBS containing 0.1% Triton X-100, 50 μg/mL RNase, and 50 μg/mL propidium iodide. Samples were analyzed by, flow cytometry and the hypodiploid peak constituted the apoptotic population in this analysis. DNA content was measured on a FACSort flow cytometer (Becton Dickinson, Franklin Lakes, NJ), and data were analyzed using ModFit software (Verity Software House, Topsham, ME). At least 1x106 cells were analyzed per sample. For Annexin V Labeling, samples were stained with fluorescein-labeled Annexin V and propidium iodide (Trevigen) according to the manufacturer’s protocol. The two Annexin V positive quadrants of the analysis were taken as the apoptotic fraction. Anoikis Assays were carried out as described previously [39 (link)]. To perform the analysis of cell surface markers (CD44+/CD24-, CD49+/CD24-), MCF10A cells were detached with trypsin, washed in blocking buffer (PBS containing 3% FBS), then stained with anti-CD24-PE (BD Biosciences, San Diego, CA, USA) and anti-CD44-PE-Cy5 (BD Biosciences) or anti-CD49-FITC (BD Biosciences) using 1 ml of antibody per 106 cells, and incubated at room temperature for 1 hr. Following incubation, cells were washed twice with 1 ml PBS. Cells were re-suspended in 1 ml PBS and then analyzed.
Maimouni S., Lee M.H., Sung Y.M., Hall M., Roy A., Ouaari C., Hwang Y.S., Spivak J., Glasgow E., Swift M., Patel J., Cheema A., Kumar D, & Byers S. (2019). Tumor suppressor RARRES1 links tubulin deglutamylation to mitochondrial metabolism and cell survival. Oncotarget, 10(17), 1606-1624.
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