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Flow cytometry buffer

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Flow cytometry buffer is a specialized liquid solution designed to support the operation and maintenance of flow cytometry equipment. It assists in the preparation, analysis, and preservation of biological samples for flow cytometry applications.

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Lab products found in correlation

Dnase, by Thermo Fisher Scientific (4 mentions) Ec800 cell analyzer, by Sony (3 mentions) 7 aminoactinomycin d, by Thermo Fisher Scientific (1 mentions) Homogenization solution, by Promega (1 mentions) Accumax, by Merck Group (1 mentions) Aria 2, by BD (1 mentions) 7 aminoactinomycin d, by BD (1 mentions) Macsquant software, by Miltenyi Biotec (1 mentions) Macsquant 10 flow cytometer, by Miltenyi Biotec (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Merck Group (1 mentions) Cd29 pe, by Thermo Fisher Scientific (1 mentions) Cd105 pe, by Thermo Fisher Scientific (1 mentions) Cd90 pe, by Thermo Fisher Scientific (1 mentions) Cd146 pe, by Thermo Fisher Scientific (1 mentions) Cd44 pe, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Flow cytometry staining buffer (22 citations) Flow Cytometry Fixation Buffer (15 citations) Flow Cytometry Permeabilization/Wash Buffer I (5 citations) Flow Cytometry Permeabilization buffer (4 citations) Flow Cytometry Fixation & Permeabilization Buffer Kit I (3 citations) Flow Cytometry Staining Buffer (1X) (3 citations) Flow cytometry fixation and permeabilization buffer kit I (2 citations) Flow Cytometry Fixation/Permeabilization Buffer I (2 citations) Flow Cytometry Human Lyse Buffer (2 citations)

7 protocols using flow cytometry buffer

1

Phenotypic Characterization of Cells

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Cells were trypsinised, counted and re-suspended in flow cytometry buffer (R & D systems) for 15 minutes and approximately 1X105 cells aliquoted into 1.5 mL microcentrifuge tubes, centrifuged at 300 g for 10 minutes and the supernatant discarded. Phycoerythrin conjugated antibodies (CD14, CD19, CD34, CD45, CD73, CD90 and CD105, HLA-DR, IgG1 and IgG2a isotype controls (Miltenyi Biotech)) in flow cytometry buffer were used to re-suspend cell pellets followed by incubation in the dark at 4°C for 10 minutes. A 10X volume of buffer was added and cells centrifuged at 300 g for 10 minutes, supernatant discarded and the cells re-suspended in flow cytometry buffer. At least 50,000 events were acquired on a Beckton Dickinson FSC500 flow cytometer. Percentage positive events were determined using gates to exclude 99% of the appropriate isotype control events.
Dale T.P., de Castro A., Kuiper N.J., Parkinson E.K, & Forsyth N.R. (2015). Immortalisation with hTERT Impacts on Sulphated Glycosaminoglycan Secretion and Immunophenotype in a Variable and Cell Specific Manner. PLoS ONE, 10(7), e0133745.
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2

Islet Cell Isolation and FACS

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Islets were dissociated in Accumax (Sigma-Aldrich, A7089) containing 10 units/mL DNase (Invitrogen, AM2222). After filtration (35-μm strainer) and centrifugation, cells were resuspended in Flow Cytometry Buffer (R&D Systems, FC001) containing 2 units/mL DNase, 0.5 mol/L EDTA, and 7-aminoactinomycin D (1:1,000; ThermoFisher, A1310). GFP+/7-aminoactinomycin D– cells were analyzed and isolated using an Aria II (BD Biosciences) and collected in Homogenization Solution (Promega, TM351; containing 1-thioglycerol).
Stancill J.S., Cartailler J.P., Clayton H.W., O’Connor J.T., Dickerson M.T., Dadi P.K., Osipovich A.B., Jacobson D.A, & Magnuson M.A. (2017). Chronic β-Cell Depolarization Impairs β-Cell Identity by Disrupting a Network of Ca2+-Regulated Genes. Diabetes, 66(8), 2175-2187.
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3

Evaluating Fibroblast Viability on Hydrogel Coatings

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Dermal fibroblasts were harvested from skin biopsies collected from the experimental animals, as previously described [20 (link)]. Cells were cultured under controlled atmosphere (37 °C, 5% CO2, humidity) using Dulbecco’s modified Eagle medium (DMEM) enriched with 10% fetal bovine serum and 1% of antibiotic-antimycotic cocktail for cell culture (Life Technologies, Carlsbad, CA, USA). To assess the cytocompatibility of the hydrogels, fibroblasts were seeded in 6-well plates (2.5 × 105 cells per well) with uncoated meshes or meshes coated with HApN or Rif-HApN (n = 3 each). Following a 24-h incubation, cells were harvested with 0.25% trypsin-EDTA (Life Technologies) and centrifuged at 200 g for 7 min. The resulting pellets were resuspended in 2 mL of flow cytometry buffer (R&D Systems, Minneapolis, MN, USA) and centrifuged again. Cells were resuspended in 400 μL of buffer mixed with 10 μL of propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) and the rate of cell viability was determined using a MACSQuant 10 flow cytometer equipped with a 488 nm laser (Miltenyi Biotec, Bergisch Gladbach, Germany). Data were analyzed using the MACSQuant software provided by the manufacturer (Miltenyi Biotec, Bergisch Gladbach, Germany).
Pérez-Köhler B., Pascual G., Benito-Martínez S., Bellón J.M., Eglin D, & Guillaume O. (2020). Thermo-Responsive Antimicrobial Hydrogel for the In-Situ Coating of Mesh Materials for Hernia Repair. Polymers, 12(6), 1245.
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4

Cell Surface Antigen Expression Analysis

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The expression of cell surface antigens on 2-14, 2-23, and 2-52 cells was analyzed by flow cytometry. Cells (2 × 105/tube) prepared as a single cell suspension by trypsin/EDTA digestion and resuspended in flow cytometry buffer (R&D Systems) were incubated with antibodies (10 mg/ml) specific for surface markers or isotype control antibodies (10 mg/ml) on ice for 45 minutes. Anti-CD105-PE and anti-CD146-PE antibodies (eBioscience, San Diego, CA) and mouse IgG-PE isotype control were used. The cells were washed with flow cytometry staining buffer and analyzed using an EC800 cell analyzer (Sony Biotechnology, Tokyo, Japan).
Hasegawa D., Hasegawa K., Kaneko H., Yoshida S., Mitarai H., Arima M., Tomokiyo A., Hamano S., Sugii H., Wada N., Kiyoshima T, & Maeda H. (2020). MEST Regulates the Stemness of Human Periodontal Ligament Stem Cells. Stem Cells International, 2020, 9672673.
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5

Multiparametric Stem Cell Profiling

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iNC-Unt, iNC-siCont, and iNC-siPAX9 were detached with Accutase (ReproCELL) and adjusted to a density of 5 × 105 cells/tube. Next, cells were incubated with R-phycoerythrin-conjugated mouse anti-human CD 34, CD45, CD90, CD105, CD166 (BioLegend, San Diego, CA, USA), IgG1, or IgG2a (iCyte, Tokyo, Japan) for 45 min at 4 °C. Cells were then washed with flow cytometry buffer (R&D Systems, Minneapolis, MN, USA) and percentages of positive cells were measured by flow cytometry (EC800 Cell Analyzer; Sony, Tokyo, Japan). Data were analyzed using Eclipse software (Sony, Tokyo, Japan).
Sugiura R., Hamano S., Tomokiyo A., Hasegawa D., Yoshida S., Sugii H., Fujino S., Adachi O., Kadowaki M., Yamashita D, & Maeda H. (2022). PAX9 Is Involved in Periodontal Ligament Stem Cell-like Differentiation of Human-Induced Pluripotent Stem Cells by Regulating Extracellular Matrix. Biomedicines, 10(10), 2366.
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6

Single-cell RBC deformability analysis

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Discarded de-identified patient blood samples were obtained from the Hematology and Oncology Division of University Hospitals under institutional review board (IRB) approval. Blood samples were collected in ethylenediaminetetraacetic acid (EDTA) anticoagulant vacutainer tubes and injected into microchannels, using disposable syringes, at precisely controlled flow shear stresses by using a syringe pump (New Era Inc., Farmingdale, NY). Whole blood samples without any dilution or pre-processing were used in the experiments. First, blood samples were introduced into the channels at 15.4 dyne/cm2 until the microchannel was completely filled with blood (Fig. 1c); then, 15 μL of blood was pumped into the channel at a shear stress of 1.54 dyne/cm2. After blood flow, channels were rinsed with flow cytometry buffer (R&D Systems, Minneapolis, MN) to remove non-adherent cells. For deformability analysis of single RBCs, an imaging area with adhered RBCs was selected and controlled shear stress with stepwise increments of 1 dyne/cm2, up to 50 dyne/cm2 was applied until the detachment of the RBCs was recorded. Microfluidic system design allowed us to precisely control fluid flow shear stress in a closed system, mimicking the physiological conditions of microvasculature. All the experiments were conducted at room temperature.
Alapan Y., Matsuyama Y., Little J.A, & Gurkan U.A. (2016). Dynamic deformability of sickle red blood cells in microphysiological flow. Technology, 4(2), 71-79.
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7

Flow Cytometry Characterization of 2-14 Cells

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The expression of cell surface antigens in 2-14 cells was analyzed by flow cytometry. 2-14 cells (2 × 10 5 cells/tube), prepared as single cell suspension by trypsin/EDTA digestion and resuspension in flow cytometry buffer (R&D Systems), were incubated with antibodies (10 mg/ml) specific for surface markers or isotype control antibodies (10 mg/ml) on ice for 45 min. Antibodies reactive to CD29-PE, CD44-PE, CD90-PE, CD105-PE, CD146-PE (eBioscience, San Diego, CA, USA), and mouse IgG isotype control-PE, were used. The cells were washed with flow cytometry staining buffer and analyzed using an EC800 cell analyzer (Sony Biotechnology, Tokyo, Japan).
Arima M., Hasegawa D., Yoshida S., Mitarai H., Tomokiyo A., Hamano S., Sugii H., Wada N, & Maeda H. (2019). R-spondin 2 promotes osteoblastic differentiation of immature human periodontal ligament cells through the Wnt/β-catenin signaling pathway. Journal of periodontal research, 54(2).
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