Cellometer cell counter

Manufactured by Revvity

Sourced in United States

The Cellometer cell counter is a laboratory instrument designed to count and analyze cells. It utilizes automated image-based detection and counting to provide accurate cell concentration measurements.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Fv1000, by Olympus (1 mentions) Histopaque 1083, by Merck Group (1 mentions) Anti biotin macsibead particles, by Miltenyi Biotec (1 mentions) Macsimag separator, by Miltenyi Biotec (1 mentions) Texmacs medium, by Miltenyi Biotec (1 mentions) Starch, by Merck Group (1 mentions) Epoch 2 plate reader, by Agilent Technologies (1 mentions) M1778, by Merck Group (1 mentions) Tamarind xyloglucan, by Megazyme (1 mentions)

9 protocols using cellometer cell counter

Cell Counting with Cellometer

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Cells were counted with the aid of a Cellometer Cell Counter (Nexcelom Bioscience).

Ratajczak H.V, & Sothern R.B. (2015). Measurement in saliva from neurotypical adults of biomarkers pertinent to autism spectrum disorders. Future Science OA, 1(4), FSO70.

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Yeast Knockout Collection Strain Culture

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The wild‐type strain BY4742 (Mat α, his3Δ1, leu2Δ0, lys2Δ0, ura3Δ0) and its isogenic derivatives deleted for SSA1 (YAL005C), SSB1 (YDL229W), and CPR6 (YLR216C) were purchased from the Thermo Yeast Knockout (YKO) Collection. Strains were grown in Yeast Nitrogen Base (YNB) medium (20 g/L glucose) supplemented with amino acids arginine (10 mL/L), leucine (20 mL/L), uracil (10 mL/L), histidine (3 mL/L) and lysine (10 mL/L) on a shaker (30°C, 300 rpm) in biological quadruplicates. The WT strain was SILAC‐labelled by growing in YNB medium supplemented with ‘light’ amino acids leucine (20 mL/L), uracil (10 mL/L), histidine (3 mL/L) and ‘heavy’ ¹³C₆‐lysine (10 mL/L) and ¹³C₆‐arginine (10 mL/L). Cells were harvested in exponential growth phase at OD₆₀₀ = 2 (± 10%) by centrifugation at 4000 rpm for 10 min at 4°C, aliquoted and stored at –80°C until further use. Cells were counted for each biological replicate using a Cellometer cell counter (Cellometer AUTOM10 by Nexcelom. http://www.nexcelom.com).

Jarnuczak A.F., Eyers C.E., Schwartz J., Grant C.M, & Hubbard S.J. (2015). Quantitative proteomics and network analysis of SSA1 and SSB1 deletion mutants reveals robustness of chaperone HSP70 network in Saccharomyces cerevisiae. Proteomics, 15(18), 3126-3139.

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Cell Growth Inhibition by JQ1

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HeLa, HCT116, and RNase H-inducible HeLa cells were treated with DMSO or 500 nM JQ1 for 72 h. Cells were harvested and cell counts performed from 3 separate experiments using a Nexcelom cellometer cell counter.

Lam F.C., Kong Y.W., Huang Q., Vu Han T.L., Maffa A.D., Kasper E.M, & Yaffe M.B. (2020). BRD4 prevents the accumulation of R-loops and protects against transcription–replication collision events and DNA damage. Nature Communications, 11, 4083.

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Measurement of Cell Dimensions

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Cell diameters of live REF52 intact cells and cytoplasts were measured using a Cellometer cell counter (Nexcelom). For volume measurements, cells were suspended in PBS (Ca²⁺/Mg²⁺ free) and stained with calcein-AM and Hoechst 33342 dyes for 15 min at 37°C. Cells were seeded on glass-bottomed culture dishes that were coated with 0.5% BSA. Cells sedimented to the glass bottom and remained nonadherent. Using an Olympus FV1000 with a 40× objective, confocal fluorescent image stacks were generated for mixed populations of intact cells and cytoplasts. Image stacks were analyzed in ImageJ based on 3D projections of masked calcein-AM and Hoechst 33342 channels. Voxels were measured for each masked image.

Graham D.M., Andersen T., Sharek L., Uzer G., Rothenberg K., Hoffman B.D., Rubin J., Balland M., Bear J.E, & Burridge K. (2018). Enucleated cells reveal differential roles of the nucleus in cell migration, polarity, and mechanotransduction. The Journal of Cell Biology, 217(3), 895-914.

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Paclitaxel Resistance in T47D Cells

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T47D_PAC cells were derived from T47D_PAR cells via continuous exposure of these cells to increasing doses of paclitaxel. The medium was replaced with fresh drug-containing medium 2-3 times a week. Resistance to 5 nM paclitaxel was observed after 6 months, and the cells were allowed to recover from treatment in drug-free medium for at least 1 month before experiments were performed. To confirm the acquisition of a resistant phenotype in T47D_PACcells, we seeded both T47D_PAR and T47D_PACcells in 6-well plates at a density of 50,000 cells/well. After 24 hours, both T47D_PAR and T47D_PACcells were treated with DMSO (control) or 2 nM paclitaxel for 6 days. Cells were counted using a Cellometer cell counter (Nexcelom Bioscience) for 6 consecutive days. For clonogenic assays, cells were seeded in 60-mm dishes at a density of 1000 cells/dish and treated with 2 nM paclitaxel for 96 hours. Drug-containing medium was then removed, and the cells were allowed to recover in drug-free medium. Dishes were scanned and the number of colonies was observed after 12 days.

Trapé A.P., Liu S., Cortes A.C., Ueno N.T, & Gonzalez-Angulo A.M. (2016). Effects of CDK4/6 Inhibition in Hormone Receptor-Positive/Human Epidermal Growth Factor Receptor 2-Negative Breast Cancer Cells with Acquired Resistance to Paclitaxel. Journal of Cancer, 7(8), 947-956.

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Quantify HCT116 Cell Viability

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HCT116 cells were transfected with miR-202-5p mimics or inhibitors for the indicated time period. The cells were then harvested using trypsin and centrifuged at 500 rpm for 3 minutes. Then the cells were re-suspended in fresh medium followed by gentle pipetting to break up clumps. The cell viability was measured via the cell counting assay using a Cellometer Cell Counter (Nexcelom Bioscience, U.S.A.).

Huang J., Zhang Y., Xu Y., Xie Q., Wu S., Dong Y., Chen B., Xia Y., Guo L., Li Q., Gu H, & Hu W. (2021). MiRNA-202-5p promotes Colorectal Carcinogenesis through suppression of PTEN. Journal of Cancer, 12(11), 3154-3163.

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Isolation of Human PBMCs

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Heparinised blood was diluted with PBS at a ratio of 1:1 and overlaid onto Histopaque 1083 (Sigma-Aldrich, Merck) density gradient medium using SepMate™-50 centrifugation tubes (Stem Cell Technologies). Samples were centrifuged at 1500 x g for 30 min at 20°C with the brake off. Peripheral blood mononuclear cells (PBMCs) were aspirated from the interface and washed twice with PBS centrifuging at 1000 x g for 10 min at 20°C. After the final wash, PBMCs for T-cell ELISpot analysis were resuspended in 3mL in RPMI 1640 medium GlutaMAX™ (ThermoFisher Scientific) only and those for B-cell ELISpot analysis were resuspended in 3mL RPMI 1640 medium GlutaMAX™ (ThermoFisher Scientific) supplemented with 10% heat-inactivated horse serum (Gibco™) and 100IU/mL penicillin and 100µg/mL streptomycin (ThermoFisher Scientific). Viable cells were counted using a Cellometer cell counter (Nexcelom Bioscience).

Fay P.C., Wijesiriwardana N., Munyanduki H., Sanz-Bernardo B., Lewis I., Haga I.R., Moffat K., van Vliet A.H., Hope J., Graham S.P, & Beard P.M. (2022). The immune response to lumpy skin disease virus in cattle is influenced by inoculation route. Frontiers in Immunology, 13, 1051008.

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Optimizing T-cell Activation and Expansion

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Cells that had been previously stored in freezing media were washed with RMPI and then centrifuged at 300 × g at 4°C for 10 min to dilute and remove any residual DMSO prior to resuspending in TexMACS medium (Miltenyi, San Diego, CA, USA), IL-2 (1.022 ng/mL), and 5% Pen-Strep. A bead-antibody cocktail was prepared by combining CD2-biotin, CD3-biotin, and CD28-biotin with anti-biotin MACSiBead particles (Miltenyi Biotec, San Diego, CA, USA) and added to the leukocyte suspension. The cell-bead mixture was plated in 6-well tissue culture-treated plates at a cell density of 1.2 × 10⁵ cells/mL and incubated at 37°C for 24 h. Following incubation, cells were removed from the 6-well plates using trypsin and a cell scraper, and 20 μL of cells were stained with acridine orange and propidium iodide (AOPI) and counted using as Cellometer cell counter (Nexcelom Bioscience, Lawrence, MA, USA). Cells were centrifuged at 300 × g at 4°C for 10 min and resuspended at a density of 2 × 10⁷ cells/mL before being placed in the MACSiMAG Separator (Miltenyi Biotec, San Diego, CA, USA) to remove the MACSiBead particles from the cell suspension prior to adoptive transfer.

Reeve K.E., Deer E., Amaral L.M., Cornelius D.C., Herrock O., Harmon A.C., Campbell N., Fitzgerald S., Ibrahim T., Wallukat G., Dechend R, & LaMarca B. (2022). Placental CD4+ T cells from preeclamptic patients cause autoantibodies to the angiotensin II type I receptor and hypertension in a pregnant rat model of preeclampsia. Exploration of medicine, 3(1), 99-111.

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Culturing Human and Mouse Bacteroides

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Bacteroides ovatus (MDA-HVS BO001) was isolated and cultured from healthy volunteer’s stool samples in a Whitley anaerobic chamber (10% H₂, 5% CO₂ and 85% N₂). Human-derived B. ovatus (ATCC 8483) and human-derived B. theta (ATCC 29148) were purchased from American Type Culture Collection (ATCC). Mouse-derived BT (MDA-JAX BT001) was previously isolated^{15 (link)}. Bacterial number was quantified using a Nexcelom Cellometer cell counter with SYTO BC dye and propidium iodide. Bacterial growth experiments were performed in a liquid media, BYEM10, composed of a hybrid of BHI and M10 supplemented with yeast extract as previously described^{15 (link),45 (link)}. Bacteria were cultured up to 24 or 48 hours at a starting concentration of 1 × 10⁶ bacteria/ml in BYEM10 broth (pH 7.2) with or without 5 mg/ml of porcine gastric mucin (M1778, Sigma-Aldrich), wheat arabionoxylan (wheat flour; low viscosity; Megazyme), xylan (Beechwood; Megazyme), xyloglucan (Tamarind; Megazyme), or starch (wheat; Sigma-Aldrich). Optical densities (OD_{600 nm}) of bacterial cultures were measured with a BioTek Epoch 2 plate reader.

Hayase E., Hayase T., Mukherjee A., Stinson S.C., Jamal M.A., Ortega M.R., Sanchez C.A., Ahmed S.S., Karmouch J.L., Chang C.C., Flores I.I., McDaniel L.K., Brown A.N., El-Himri R.K., Chapa V.A., Tan L., Tran B.Q., Pham D., Halsey T.M., Jin Y., Tsai W.B., Prasad R., Glover I.K., Ajami N.J., Wargo J.A., Shelburne S., Okhuysen P.C., Liu C., Fowler S.W., Conner M.E., Peterson C.B., Rondon G., Molldrem J.J., Champlin R.E., Shpall E.J., Lorenzi P.L., Mehta R.S., Martens E.C., Alousi A.M, & Jenq R.R. (2023). Bacteroides ovatus alleviates dysbiotic microbiota-induced intestinal graft-versus-host disease. Research Square.

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