Facsscan flow cytometer

Manufactured by BD

Sourced in United States, Germany

The FACSscan flow cytometer is a laboratory instrument used for the analysis of cells or other particles in a fluid suspension. It is designed to measure and analyze the physical and fluorescent characteristics of these particles as they pass through a laser beam. The FACSscan is capable of detecting and quantifying multiple parameters simultaneously, providing researchers with detailed information about the properties of the samples under investigation.

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Lab products found in correlation

Facsdiva software, by BD (8 mentions) Fitc annexin 5 apoptosis detection kit, by BD (4 mentions) Cellquest software, by BD (3 mentions) Propidium iodide, by Thermo Fisher Scientific (3 mentions) Modfit software, by Verity Software House (2 mentions) Fitc labeled annexin 5 and pi, by BD (2 mentions) Alexa fluor 488 annexin 5 dead apoptosis kit, by Thermo Fisher Scientific (2 mentions) Axiovert 200m, by Zeiss (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (2 mentions) Flow cytometer facs aria, by BD (2 mentions) Cytomics fc500 flow cytometer, by Beckman Coulter (2 mentions) Vegfr2 antibody, by BD (2 mentions) Fcr blocking reagent, by Miltenyi Biotec (2 mentions) Annexin 5 fitc apoptosis detection kit, by Enzo Life Sciences (2 mentions) Propidium iodide, by BD (2 mentions) Rf 5301pc spectrofluorophotometer, by Shimadzu (1 mentions) Cellquest pro software, by BD (1 mentions) Anti hla dr, by BD (1 mentions) Anti cd8, by BD (1 mentions) Anti cd19, by BD (1 mentions)

71 protocols using facsscan flow cytometer

Cell Cycle Analysis of ZINC69391 Treatment

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Synchronized cell populations were treated with 50 μM of ZINC69391 or 0.05% (v/v) DMSO (vehicle) for 15h. After treatment, cells were harvested, washed with ice-cold PBS, fixed overnight by addition of 70% (v/v) ethanol and stored at -20°C for a minimum of 24h. On the day of flow cytometry analysis, cell suspensions were washed with ice-cold PBS and re-suspended in 50 μl RNase A (100 μg/ml) at room temperature for 15 min.
Propidium Iodide was added to a final concentration of 20 μg/ml and incubated in dark at room temperature for 20 min. Cell cycle phase distributions were analyzed by FACS Scan Flow Cytometer (Beckton-Dickinson CA, USA). Data from at least three independent experiments were analyzed using ModFit software (VeritySoftware House Inc., Topsham, ME, USA) to determine the fractions of cells in the subG0/G1, G0/G1, S and G2/M phases from cell cycle distribution.

Cabrera M., Echeverria E., Lenicov F.R., Cardama G., Gonzalez N., Davio C., Fernández N, & Menna P.L. (2017). Pharmacological Rac1 inhibitors with selective apoptotic activity in human acute leukemic cell lines. Oncotarget, 8(58), 98509-98523.

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Mitochondrial Membrane Potential Assay

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The ΔΨm was determined in phosphate based-buffer, pH 7.4. Washed platelets (1 mg protein/mL) were incubated at 37°C with 0.25 μM rhodamine 6G [19 ] or 2 μM 5,5´,6,6-tetrachloro-1,1´,3,3´-tetraethyl-benzimidazolyl-carbocyanine iodide (JC-1) [20 (link)]. For rhodamine, fluorescence was registered in a RF 5301PC spectrofluorophotometer (Shimadzu Scientific Instruments, Maryland, USA) using 480 nm and 565 nm for excitation and emission wavelengths, respectively. Maximal fluorescence indicating total ΔΨm was attained for each experiment after adding 2.5 μM CCCP. For JC-1 assays, washed platelets (1x10⁴ platelets/μL) were analyzed in a FACS Scan flow cytometer (Beckton Dickinson, San Jose, CA) at 37°C, the excitation wavelength was 488 nm and the emission wavelengths were 535 nm for green (FL1) and 590 nm for red (FL2). Following 2.5 μM CCCP treatment the mitochondrial membrane potential of the cells was eliminated, as demonstrated by the increase of the monomer emission [20 (link)].

Corona de la Peña N., Gutiérrez-Aguilar M., Hernández-Reséndiz I., Marín-Hernández Á, & Rodríguez-Enríquez S. (2017). Glycoprotein Ib activation by thrombin stimulates the energy metabolism in human platelets. PLoS ONE, 12(8), e0182374.

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Cell Cycle Analysis by PI Staining

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Cell cycle was analyzed by Propidium Iodide (PI) staining. Briefly, 1,000,000 cells were fixed with 2 ml cold ethanol for 1 hour at 4°C. After washed with ice-cold PBS for two times, the cells were suspended with 1 ml of PI staining solution (40 μg/ml in PBS), supplemented with 50 μl of RNase A stock solution (10 μg/ml) and incubated 2 hour at 4°C. Then cell cycle was analyzed using FACSScan flow cytometer (Becton Dickinson, San Jose, CA).

Gu C., Peng H., Lu Y., Yang H., Tian Z., Yin G., Zhang W., Lu S., Zhang Y, & Yang Y. (2017). BTK suppresses myeloma cellular senescence through activating AKT/P27/Rb signaling. Oncotarget, 8(34), 56858-56867.

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Phenotypic Characterization of AT-MSCs

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AT-MSCs (10⁶) were trypsinized and incubated with fluorescein isothiocyanate (FITC)-conjugated CD34, CD45, CD90, phycoerythrin (PE) -conjugated human leukocyte antigen (HLA)-DR, CD11b, CD29, CD105 (mouse, monoclonal, 1:200; eBioscience, San Diego, CA, USA) or PE-conjugated CD3, CD73, CD117, (mouse, monoclonal, 1:200; BD Biosciences, San Jose, CA, USA) antibodies for 30 minutes at RT, followed by three washes. Staining with unconjugated anti-programmed death ligand (PDL) 1 (rabbit, polyclonal, 1:100; BD Biosciences), FITC-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (Ab) (goat, polyclonal, 1:200; BD Biosciences) was used as a secondary antibody. The fluorescent labeled cells were analyzed on a FACSscan flow cytometer (Becton-Dickinson, Franklin Lakes, NJ, USA) using CELLQuest Pro software (Becton-Dickinson).

YIN L., ZHU Y., YANG J., NI Y., ZHOU Z., CHEN Y, & WEN L. (2014). Adipose tissue-derived mesenchymal stem cells differentiated into hepatocyte-like cells in vivo and in vitro. Molecular Medicine Reports, 11(3), 1722-1732.

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Cell Cycle Analysis by Flow Cytometry

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CT26 cells (1 × 10⁶) were harvested and washed twice with cold phosphate-buffered saline (PBS). Cells then were stained with 500 µl of propidium iodide (PI) staining solution (50 mg/ml PI, 0.1% Triton X-100, 200 mg/ml DNase-free RNase in PBS) for 30 min at room temperature in the dark. Ten thousand events per sample were acquired using a FACS-scan flow cytometer (Becton-Dickinson, San Jose, CA, USA), and the percentage of cells in G₀/G₁, S, G₂/M, and sub-G₂/M phases of the cell cycle were determined using CELL Quest software (Becton-Dickinson).

Zhou C., Zhang P., Xu G.C., Wu D.M., Liu R.Y., Zeng Q, & Wang C.T. (2015). RNA Interference of Biot2 Induces G1 Phase Arrest and Apoptosis in Mouse Colorectal Cancer Cell Line. Oncology Research, 22(2), 93-103.

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Immunophenotypic Analysis of Peripheral Lymphocytes in COVID-19 Patients

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Peripheral blood white blood cells (WBC) of all participants were isolated from their hemogram tubes with EDTA taken for their routine tests by using erythrocyte lysing solution (155 mM NH4Cl; 10 mM KHCO3; 0,1 mM EDTA; pH:7.3). The following fluorochrome labeled monoclonal antibodies (mAb) and isotype-matched controls were used for two–three color phenotypic analysis: anti-IgG1, anti-IgG2a, anti-CD45, anti-CD3; anti-CD4; anti-CD8; anti-CD16; anti-CD56; anti-CD19; anti-HLA-DR; anti-CD69, (Becton&Dickinson Corp, San Jose, CA, USA). Cells were acquired and analyzed using CellQuest software on a FACS scan flow cytometer (Becton Dickinson Inc, San Jose, CA, USA). Lymphocytes were gated according to their forward and side scatter characteristics and their specific CD markers. Markers were evaluated as percentages. In 13 of 20 patients, peripheral lymphocyte subset analysis was repeated 1 week later. A total of 10 KTR without COVID-19 was used as control group.

Ozkok A., Alpay N., Alan S., Bakan N.D., Soysal F., Yazici H., Ekşioğlu-Demiralp E, & Yildiz A. (2021). Immunological parameters associated with the severity of COVID-19 pneumonia in kidney transplant recipients. International Urology and Nephrology, 54(5), 1105-1116.

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Cell Cycle Analysis of Fe3O4@Glu/BTSC

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The effect of Fe₃ O₄@Glu/BTSC on G0/G1, S and G2/M phases was evaluated according to the method described by Davy and Doorbar.^{26 (link)} After culture, the cells were treated with concentrations of 10–20 μL of Fe₃ O₄@Glu/BTSC and 0.05% DMSO (as negative control) for 15 hours. Rinsing was performed with PBS and kept in a -20°C freezer for 24 hours with the addition of 70% alcohol overnight. To perform flow cytometry, the cells were washed again with cold PBS and suspended for 15 minutes at room temperature by adding 50 µL of RNase A 100 μg/mL. In the next step, propidium iodide was added to the suspension to reach the final concentration of 20 μg/mL. The suspension was placed at room temperature for 20 minutes. Cell cycle phases were analyzed by FACS Scan Flow Cytometer (Becton-Dickinson CA, USA). Data were analyzed with three replications, and different phases of the cell cycle were analyzed using flowing software 2.5.1.

Habibi A., Bakhshi N., Moradi shoili Z, & Amirmozafari N. (2022). Iron Oxide Nanoparticles Conjugated to Thiosemicarbazone Reduce the Survival of Cancer Cells by Increasing the Gene Expression of MicroRNA let-7c in Lung Cancer A549 Cells. Archives of Iranian medicine, 25(12), 807-816.

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Apoptosis Assay of Drug-Treated Cells

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Cells were seeded into 6-well plates and incubated for 48 hours with a drug-containing medium when the cell confluency reached 70%. Then, 1 × 10⁶ cells were collected and stained according to the steps of the apoptosis kit instructions, and the percentage of apoptotic cells was determined using a FACSscan flow cytometer (Becton-Dickinson, NJ, USA).

Li H., Xiao J., Li X., Huang Q., Liu Q, & Zhang Q. (2023). Mechanism of morusin on breast cancer via network pharmacology and in vitro experiments. Medicine, 102(28), e34300.

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Receptor Expression in Diabetic Cells

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To assess the surface expression of receptors, CD34⁺ cells from diabetics and non-diabetics were treated with FcR blocking reagent (anti-CD16/32, TruStain, BioLegend) and then stained with Zenon Alexa Fluor 488-(Invitrogen) tagged primary mouse monoclonal anti-human VEGFR1, antihuman VEGFR2, and CXCR4 antibodies (R&D systems), or isotype control antibodies for 20 minutes on ice. Cells were analyzed by a FACS Scan flow cytometer (Becton Dickinson).

Jarajapu Y.P., Hazra S., Segal M., LiCalzi S., Jhadao C., Qian K., Mitter S.K., Raizada M.K., Boulton M.E, & Grant M.B. (2014). Vasoreparative Dysfunction of CD34+ Cells in Diabetic Individuals Involves Hypoxic Desensitization and Impaired Autocrine/Paracrine Mechanisms. PLoS ONE, 9(4), e93965.

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Par-4 Mutants Modulate TRAIL-Induced Apoptosis

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Cells transfected with Par-4 mutants were either treated with recombinant His-TRAIL (500 ng/ml) or Paclitaxel 200 nM (Calbiochem). Cells were harvested and fixed with 0.2% paraformaldehyde in presence of 1 mg/ml of DAPI or Hoetsch 33342 (Sigma-Aldrich) to stain the nuclei. Apoptosis was measured by counting cells that are condensed and fragmented. Images were acquired using the Cell Observer station (Zeiss, Göttingen, Germany). Briefly, the system consists of an inverted microscope AxioVert 200M (Zeiss) equipped for fluorescence with a CCD camera. All the system is motorized and controlled by the Axiovision software (Zeiss). Apoptosis was also assessed by the determination of PARP and caspase 8 cleavage by western blotting using anti-mouse caspase-8 from R&D and rabbit PARP (detection of the short cleaved form) or mouse PARP (detection of the large fragment) antibodies (Santa Cruz).
For FACS analysis, cells stained with APO and FITC–annexin V conjugate were analyzed by flow cytometry using a FACS Scan flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). Caspase-3 activity was measured by FACS using the fluorochrome benzyloxycarbonyl-Asp-Glu-Val-Asp (OMe) fluoromethylketone (PromoKine Caspase-3 kit, PromoCell, Heidelberg, Germany).

de Thonel A., Hazoumé A., Kochin V., Isoniemi K., Jego G., Fourmaux E., Hammann A., Mjahed H., Filhol O., Micheau O., Rocchi P., Mezger V., Eriksson J.E., Rangnekar V.M, & Garrido C. (2014). Regulation of the proapoptotic functions of prostate apoptosis response-4 (Par-4) by casein kinase 2 in prostate cancer cells. Cell Death & Disease, 5(1), e1016-.

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