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Coulter counter analyzer

Manufactured by Beckman Coulter
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The Coulter Counter analyzer is a lab equipment designed to count and size particles suspended in a conductive liquid. It uses the principle of electrical impedance to detect and measure the size and number of particles, such as cells or other microscopic entities, within a sample. The Coulter Counter provides accurate and reliable particle counting and sizing data to support various laboratory applications.

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Lab products found in correlation

Vacutainer system, by BD (3 mentions) Prism 5, by GraphPad (2 mentions) Vi cell xr cell viability analyzer, by Beckman Coulter (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Gsk126, by Active Motif (1 mentions)

10 protocols using coulter counter analyzer

1

Quantifying Cell Proliferation via Trypan Blue

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The effect of miR-9 on cell growth was determined through proliferation assays. M12 and M12 cells transformed with the miR-9 inhibitor plasmid were plated (5 x 103 cells) onto a 24-well plate. The adherent cells were released by incubating with 0.25% Trypsin-EDTA (Gibco-Life Technologies) after which the trypsin was inactivated by washing the cells in serum-containing media. The cells counted using a Coulter Counter® Analyzer (Beckman Coulter, Inc) or Vi-Cell™ XR Cell Viability Analyzer (Beckman Coulter, Inc) using a trypan blue solution. Cells were >90% live when analyzed in this fashion.
Seashols-Williams S.J., Budd W., Clark G.C., Wu Q., Daniel R., Dragoescu E, & Zehner Z.E. (2016). miR-9 Acts as an OncomiR in Prostate Cancer through Multiple Pathways That Drive Tumour Progression and Metastasis. PLoS ONE, 11(7), e0159601.
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2

Healthy Adult Blood Collection Protocol

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Our study protocol was evaluated by the local medical ethical board (Medical Ethical Committee of Maastricht University Medical Center). The study population consisted of 129 healthy adult individuals, aged 20–65 years. All participants did not take any oral anticoagulant or anti-platelet drugs for at least two weeks, did not have a history of thrombosis or bleeding and gave full written informed consent according to the Helsinki declaration. Experiments were conducted at the Synapse Research Institute in accordance with approved guidelines and regulations. Blood was collected aseptically by antecubital puncture via a 21-gauge needle (0.8x32 mm) into vacuum tubes (1 volume trisodium citrate 0.105M to 9 volumes blood) (BD Vacutainer System/Greiner). The blood was kept at RT and used within 4 hours from withdrawal. Cell counts in whole blood were performed with a Coulter Counter analyzer (Beckman Coulter, Woerden, The Netherlands).
Huskens D., Sang Y., Konings J., van der Vorm L., de Laat B., Kelchtermans H, & Roest M. (2018). Standardization and reference ranges for whole blood platelet function measurements using a flow cytometric platelet activation test. PLoS ONE, 13(2), e0192079.
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3

Cell Viability Assay with DAC and GSK-126

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SNU398, HepG2 and SNU475 cells were obtained from the American Type Culture Collection (ATCC; www.atcc.org). Each cell line was treated with daily doses of 100 nM DAC (Sigma-Aldrich) and/or 500 nM GSK-126 (Active Biochem, LTD, Hong Kong) for a total of three days. Treated cells continued to grow after the drug(s) were removed. SNU398 and HepG2 cells were harvested on day 14 after treatment, while SNU475 cells were harvested on day 24. Cell survival rates were assessed on specific days using a Coulter counter analyzer (Beckman Coulter, Inc.). Results are normalized to control group (cells treated with phosphate buffered saline (DPBS)). The data are presented as means of triplicate measurements; error bars represent standard errors of the mean (SEMs). The cell doubling times were calculated as described below: Doubling time = A*log(2)/(log(B) − log(C)), in which A is the number of days from cell seeded to harvested, B is the total number of harvested cells, and C is the number of seeded cells.
Zhang L., Li H.T., Shereda R., Lu Q., Weisenberger D.J., O’Connell C., Machida K., An W., Lenz H.J., El-Khoueiry A., Jones P.A., Liu M, & Liang G. (2022). DNMT and EZH2 inhibitors synergize to activate therapeutic targets in hepatocellular carcinoma. Cancer letters, 548, 215899.
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4

Healthy Adult Platelet Characterization

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Our study protocol was evaluated by the local medical ethical board (Medical Ethical Committee of Maastricht University Medical Center). Blood was collected into vacutainer tubes (with 3.2% sodium citrate; from BD Vacutainer System) from healthy adults who gave full informed consent according to the Helsinki declaration and had not taken any anticoagulants or platelet inhibitors for at least 2 weeks and had no history of thrombosis or bleeding. Blood was kept at room temperature and used within 4 hours after collection. Cell counts were measured on a Coulter Counter analyzer (Beckman Coulter).
Wan J., Konings J., Yan Q., Kelchtermans H., Kremers R., de Laat B, & Roest M. (2020). A novel assay for studying the involvement of blood cells in whole blood thrombin generation. Journal of Thrombosis and Haemostasis, 18(6), 1291-1301.
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5

Cell Fractionation and TNFα Stimulation

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A cell fractionation kit (Cell Signaling Technology) was used to collect whole cell lysate (WCL), cytoplasmic (cyto), and nuclear (nuc) fractions from specified cell populations. Briefly, OciLy3 cells were treated with 50 ng of recombinant TNFΑα (Fisher) or PBS control for 1.5 hours in regular growth media. Cells were counted using a Coulter Counter analyzer (Beckman). 5 × 106 cells per group were collected, and cell fractionation was performed per manufacturer specifications.
Yushi Q., Li Z., Von Roemeling C.A., Doeppler H., Marlow L.A., Kim B.Y., Radisky D.C., Storz P., Copland J.A, & Tun H.W. (2016). Osteopontin is a multi-faceted pro-tumorigenic driver for central nervous system lymphoma. Oncotarget, 7(22), 32156-32171.
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6

Cell Proliferation Assay for Synergistic Effects

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786-0 (3×10 4 ), ACHN (1.5×10 5 ), A-498 (3×10 4 ), Caki-2 (1×10 5 ) and 769-P (4×10 4 ) cells were seeded into 6well tissue culture plates, treated with the indicated compounds or vehicle control, and were harvested, resuspended, and counted using a coulter counter analyzer (Beckman Coulter, Inc.) as indicated in Figure 1. The synergistic effect of DAC and BMN-673 was evaluated using CompuSyn software (https://www.combosyn.com) (Supplemental Table 1). Combination index (CI)<1 indicates a synergistic effect, CI=1 indicates an additive effect and CI>1 indicates antagonistic effect.
Zhou X., Sekino Y., Li H.T., Fu G., Yang Z., Zhao S., Gujar H., Zu X., Weisenberger D.J., Gill I.S., Tulpule V., D'souza A., Quinn D.I., Han B, & Liang G. (2023). SETD2 Deficiency Confers Sensitivity to Dual Inhibition of DNA Methylation and PARP in Kidney Cancer. Cancer research, 83(22).
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7

Simultaneous Blood Sample Collection

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Two simultaneous blood samples were drawn from the cubital vein and the right internal jugular vein at the end of the HVPG procedure 22 into tubes containing 3.2% sodium citrate (#454332, Greiner Bio-one, Austria). Both samples were processed in parallel at room temperature within 30 minutes, and flow cytometry was performed within 1 hour. All pairs of samples were drawn by the same physician and analyzed by the same operator to minimize procedural deviations. All blood samples were kept at room temperature. Blood cell counts were determined by a Coulter counter analyzer (Beckman Coulter, Netherlands).
Brusilovskaya K., Simbrunner B., Lee S., Eichelberger B., Bauer D., Zinober K., Schwabl P., Mandorfer M., Panzer S., Reiberger T, & Gremmel T. (2022). Peripheral versus central venous blood sampling does not influence the assessment of platelet activation in cirrhosis. Platelets, 33(6).
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8

Healthy Adult Blood Collection Protocol

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Our study protocol was evaluated by the local medical ethical board (Medical Ethical Committee of Maastricht University Medical Center). The study population consisted of 118 healthy adult individuals, aged 20 to 65 years. All participants did not take any oral anticoagulant or antiplatelet drugs for at least 2 weeks, did not have a history of thrombosis or bleeding, and gave full informed consent according to the Helsinki Declaration. Experiments were conducted at the Synapse Research Institute in accordance with approved guidelines and regulations. Blood was collected aseptically by antecubital puncture via a 21-gauge needle (0.8 Â 32 mm) into vacuum tubes (1 volume trisodium citrate 0.105 M to 9 volumes blood) (BD Vacutainer System/Greiner). The blood was kept at RT and used within 4 hours after collection. Cell counts in whole blood were performed with a Coulter Counter analyzer (Beckman Coulter, Woerden, The Netherlands).
Sang Y., Huskens D., Wichapong K., de Laat B., Nicolaes G.A.F, & Roest M. (2019). A Synthetic Triple Helical Collagen Peptide as a New Agonist for Flow Cytometric Measurement of GPVI-Specific Platelet Activation. Thrombosis and haemostasis, 119(12).
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9

Evaluating DAC's Anticancer Effects in Renal Cell Carcinoma

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For the IC50 analysis, 786-O (100 cells), A-498 (150 cells), A-704 (600 cells), ACHN (600 cells), Caki-1 (300 cells), Caki-2 (300 cells), and RenCa (100 cells) cells were plated in each well of the 96-well plates 24 h prior to the treatment. Cells were treated with DAC (from 10 to 0.0064 μM with 5 X dilution) for 24 h (786-O and RenCa) or 48 h (A-498, A-704, ACHN, Caki-1 and Caki-2). Cell viability was measured at day 8 after treatment using the CellTiter-Glo 2.0 Cell Viability Assay. IC50 was calculated and plotted using GraphPad Prism 5.
To compare how genetic manipulation of SETD2 affected cell proliferation, 20,000 786-O cells were seeded in 6-well plates at day 0 and treated with 100 (for doubling time assay) or 300 nM (rest of experiments) DAC at day 1 for 24 h. At the indicated time points in Figure 2C, cells were harvested and counted using Coulter counter analyzer to reseed 20,000 cells into new plates for PDT calculations. PDTs were calculated using the equation: PDT = duration x LOG (2)/(LOG (final cell number) – LOG (initial cell number)).
Jones T.L., Tucker D.E, & Ort D.R. (1998). Chilling Delays Circadian Pattern of Sucrose Phosphate Synthase and Nitrate Reductase Activity in Tomato. Plant Physiology, 118(1), 149-158.
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10

Evaluating DAC's Anticancer Effects in Renal Cell Carcinoma

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For the IC50 analysis, 786-O (100 cells), A-498 (150 cells), A-704 (600 cells), ACHN (600 cells), Caki-1 (300 cells), Caki-2 (300 cells), and RenCa (100 cells) cells were plated in each well of the 96-well plates 24 h prior to the treatment. Cells were treated with DAC (from 10 to 0.0064 μM with 5 X dilution) for 24 h (786-O and RenCa) or 48 h (A-498, A-704, ACHN, Caki-1 and Caki-2). Cell viability was measured at day 8 after treatment using the CellTiter-Glo 2.0 Cell Viability Assay. IC50 was calculated and plotted using GraphPad Prism 5.
To compare how genetic manipulation of SETD2 affected cell proliferation, 20,000 786-O cells were seeded in 6-well plates at day 0 and treated with 100 (for doubling time assay) or 300 nM (rest of experiments) DAC at day 1 for 24 h. At the indicated time points in Figure 2C, cells were harvested and counted using Coulter counter analyzer to reseed 20,000 cells into new plates for PDT calculations. PDTs were calculated using the equation: PDT = duration x LOG (2)/(LOG (final cell number) – LOG (initial cell number)).
Li H.T., Jang H.J., Rohena-Rivera K., Liu M., Gujar H., Kulchycki J., Zhao S., Billet S., Zhou X., Weisenberger D.J., Gill I., Jones P.A., Bhowmick N.A, & Liang G. (2023). RNA mis-splicing drives viral mimicry response after DNMTi therapy in SETD2-mutant kidney cancer. Cell reports, 42(1), 112016.
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