Facscan analytic flow cytometer

Manufactured by BD

Sourced in United States, Canada

The FACScan Analytic Flow Cytometer is a laboratory instrument designed for the analysis of cells and particles in a fluid sample. It utilizes the principles of flow cytometry to detect and measure various characteristics of the cells, such as their size, granularity, and fluorescence properties. The core function of the FACScan is to provide researchers and scientists with a tool for high-throughput, quantitative analysis of cell populations.

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Lab products found in correlation

Modfit software, by Bio-Rad (1 mentions) Fcs express software, by De Novo Software (1 mentions) Anti cd44 fitc, by BD (1 mentions) Cd44 fitc, by BD (1 mentions) Pkh67 fluorescent dye, by Merck Group (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Jsm 6700f sem, by JEOL (1 mentions) Tcs sp mp inverted confocal microscope, by Leica (1 mentions) Pe conjugated goat anti human igg, by Jackson ImmunoResearch (1 mentions)

7 protocols using facscan analytic flow cytometer

Cell Cycle and Apoptosis Analysis

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Cells were fixed overnight at 4°C in 70% ice-cold ethanol. For cell cycle analysis, cells were stained with PI from Cell cycle kits (Beyotime, China) based on the manufacturer’s instructions. Cells were analyzed on FACScan Analytic Flow Cytometer (BD Bioscience, San Jose, California, USA). Cell cycle distribution was analyzed by using the Modfit software (Bio-Rad, Hercules, CA, USA).
For cell apoptosis analysis, cells were fixed and then stained with Annexin V-FITC from Annexin-V-FITC cell apoptosis kit (Beyotime, China) and then stained with PI. Cells were analyzed on a flow cytometer (BD Bioscience). Cell apoptotic rate was calculated by the ratio of early and late apoptotic cells to total cells.

Zhang Q., Zhang J., Zhang J., Aerxiding P., Quhai A., Chen C, & Shan L. (2019). The Biological-Behavioral Effect Of Neuritin On Non-Small Cell Lung Cancer Vascular Endothelial Cells Via VEGFR And Notch1. OncoTargets and therapy, 12, 9747-9755.

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Phenotyping 2D and 3D Cultured Cells

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Cells were harvested from both 2D (conventional culture) and 3D (scaffold) culture, pelleted via centrifugation, washed in FACS buffer (sterile PBS supplemented with 1% ES-FBS, 2% BSA, and 0.25% saponin), and stained with primary antibodies (smooth muscle marker: SM-myosin; endothelial cell marker: CD31). The cells were gated by forward scatter versus side scatter to eliminate debris. A minimum of 10,000 events was counted for each analysis. All analyses were performed using a Becton Dickinson FACS can analytic flow cytometer (BD Biosciences) with FCS Express software (DeNovo Software, Thornhill, ON, Canada) at the UCLA Flow Cytometery Laboratory.

Gluck J.M., Delman C., Chyu J., MacLellan W.R., Shemin R.J, & Heydarkhan-Hagvall S. (2014). Microenvironment influences vascular differentiation of murine cardiovascular progenitor cells. Journal of biomedical materials research. Part B, Applied biomaterials, 102(8), 1730-1739.

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Flow Cytometry Analysis of Cell Viability

Cited 2 times

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The CCL-23 and UM-SCC-1 cell lines were treated with different combinations of cisplatin, liposome, liposomal curcumin, and liposomal CDF, and stained with CD44-FITC (Cat #555478, BD Biosciences, San Jose, CA) or Annexin-PE (Oncogene Research products, Boston, MA). Intracellular P16 was stained using intracellular staining kit protocol (Cat #415, BD Biosciences, San Jose, CA) prior to staining with extracellular membrane staining with anti-CD44-FITC or anti-Annexin-PE. Stained cells were analyzed using a Becton Dickinson FACScan analytic flow cytometer (Becton Dickinson, Franklin Lakes, NJ). Samples were protected from light and maintained on ice and analyzed within an hour of staining [6 (link), 10 (link), 11 (link)].

Basak S.K., Zinabadi A., Wu A.W., Venkatesan N., Duarte V.M., Kang J.J., Dalgard C.L., Srivastava M., Sarkar F.H., Wang M.B, & Srivatsan E.S. (2015). Liposome encapsulated curcumin-difluorinated (CDF) inhibits the growth of cisplatin resistant head and neck cancer stem cells. Oncotarget, 6(21), 18504-18517.

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Evaluating anti-EMP2 and anti-HER2 effects on PBMC-mediated cytotoxicity

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Peripheral blood mononuclear cells (PBMCs) were isolated from blood using Ficoll-Paque plus (GE Healthcare, Pittsburg, PA) from three volunteers. Blood donors had given informed consent before for obtaining a peripheral venous blood sample for PBMC assays. These experiments were done according to the rules of the Ethical Committee of University of California, Los Angeles. PBMCs were resuspended in DMEM with 10% FCS. SK-BR-3 cells were initially labeled using PKH67 fluorescent dye (Sigma Aldrich) and then plated in 6 well plates. Cells were pre-incubated with anti-EMP2 IgG1 or trastuzumab (anti-HER2/neu; Genentech, San Francisco, CA) as a positive control overnight and then incubated with different ratios of PBMCs for 4–8 hours at 37°C. The percentage of cell death was quantitated by propidium iodide staining using a Becton Dickinson FACScan Analytic Flow Cytometer (Becton Dickinson) at the UCLA Jonsson Comprehensive Cancer Center (JCCC) and Center for AIDS Research Flow Cytometry Core Facility. Experiments were performed in duplicates, normalized to an untreated negative control, and then averaged.

Fu M., Maresh E.L., Helguera G.F., Kiyohara M., Qin Y., Ashki N., Daniels-Wells T.R., Aziz N., Gordon L.K., Braun J., Elshimali Y., Soslow R.A., Penichet M.L., Goodglick L, & Wadehra M. (2014). Rationale and pre-clinical efficacy of a novel anti-EMP2 antibody for the treatment of invasive breast cancer. Molecular cancer therapeutics, 13(4), 902-915.

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Characterization of Polymer Nanocapsules

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Example 11

IR spectra of the polymer nanocapsules were obtained on a PerkinElmer Paragon 1000 FT-IR spectrometer. UV-Visible spectra were acquired with a GeneSys 6 spectrometer (Thermo Scientific). Fluorescence spectra were obtained with a QuantaMaster Spectrofluorimeter (Photon Technology International). TEM images of nanocapsules were obtained on a Philips EM120 TEM at 100000× (see, e.g., FIG. 3, FIG. 4, and FIG. 5).

Before observation, siRNA nanocapsules were negatively stained using 1% pH 7.0 phosphotungstic acid (PTA) solution. Zeta potential and particle size distribution were measured with a Malvern particle sizer Nano-ZS. SEM images of nanocapsules were obtained with a JEOL JSM-6700F SEM. Dry samples on a silicon surface were sputter-coated with gold before measurement. Fluorescent images of cells were obtained with either Zeiss Axio Observer.Z1 fluorescence microscope or Leica TCS SP MP Inverted Confocal Microscope. Cellular fluorescent intensity distribution was determined with Becton Dickinson FACScan Analytic Flow Cytometer. A 488 nm argon laser was used as the excitation light.

US10568844B2. RNAi molecule delivery platform based on single-siRNA and shRNA nanocapsules (2020-02-25). THE REGENTS OF THE UNIVERSITY OF CALIFORNIA [US]. Inventors: Yunfeng Lu [US], Irvin S. Y. Chen [US], Ming Yan [US], Min Liang [US], Masakazu Kamata [US], Jing Wen [US].

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Measurement of Cellular and Mitochondrial ROS

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Cells (4x10⁵ per well) were seeded into 6-well plates in complete medium. The next day, cells were cultured with either the complete medium or medium without glucose for 4 h. For measurement of cellular ROS levels, cells were incubated with 2.5 μM DCFDA for 30 min before analysis using a Becton Dickinson FACScan analytic flow cytometer. For measurement of mitochondrial ROS levels, the cells were incubated with 2.5 μM MitoSOX for 30 min before the flow cytometer analysis. Cells were gated using forward scatter and side scatter to remove debris and dead cells, and 10⁴ live cell events were recorded. To quantify changes in DCFDA or mitoSOX signal, mean fluorescent intensity after gating was used.

Hong J., Maacha S., Pidkovka N., Bates A., Salaria S.N., Washington M.K, & Belkhiri A. (2022). AXL Promotes Metformin-Induced Apoptosis Through Mediation of Autophagy by Activating ROS-AMPK-ULK1 Signaling in Human Esophageal Adenocarcinoma. Frontiers in Oncology, 12, 903874.

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Flow Cytometry Analysis of EMP2 Expression

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EMP2-positive cells (HEC1a/EMP2, HEC1a,(15 (link)) or 4T1 cells) or EMP2-negative cells (EL4 or Ramos) were resuspended at a concentration of 10⁶ cells in 1 mL of cold PBS+0.2% BSA buffer (flow buffer). The cell suspension was centrifuged for 5 min at 500g, at 4°C. Cells were then incubated with 1 µg of recombinant anti-EMP2 IgG1 for 2 h at 4°C. An IgG specific for the hapten dansyl, 5-dimethylamino naphthalene-1-sulfonyl chloride (DNS) was used as a non-targeted antibody negative control (26 (link)). Cells were washed three times and then incubated for 30min at 4°C with PE-conjugated goat anti-human IgG (Jackson Immunoresearch). Cells were washed and resuspended in flow buffer. Flow cytometry was immediately performed with a Becton Dickinson FACScan Analytic Flow Cytometer (Becton Dickinson, Franklin Lakes, NJ) in the UCLA Jonsson Comprehensive Cancer Center and Center for AIDS Research Flow Cytometry Core Facility.

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