Coulter counter

Manufactured by Beckman Coulter

Sourced in United States, United Kingdom, Belgium, Germany, Japan, Canada, France, Italy

The Coulter Counter is a laboratory instrument used for counting and measuring the size of particles suspended in a liquid. It utilizes the Coulter principle to detect and analyze particles, providing accurate and reliable data on the particle population within a sample.

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Lab products found in correlation

Ack lysing buffer, by Lonza (7 mentions) Rat anti mouse cd16 32 antibody, by BD (5 mentions) Facscanto, by BD (5 mentions) Ficoll paque, by GE Healthcare (5 mentions) Trypsin edta, by Thermo Fisher Scientific (4 mentions) Victor x3 light plate reader, by PerkinElmer (4 mentions) 7 aad, by BD (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Edta monoject tubes, by Medtronic (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Pe cy5 conjugated anti mouse cd4, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) 24 well culture plate, by Corning (2 mentions) Gentlemacs dissociator, by Miltenyi Biotec (2 mentions) Synergy 2 multi function microplate reader, by Agilent Technologies (2 mentions) Luerlock syringes, by Cole-Parmer (2 mentions) Fetal clone 3, by GE Healthcare (2 mentions) Penicillin streptomycin mixture, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)

392 protocols using coulter counter

Compound Effects on Cell Proliferation

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HMEC-1 cells were seeded in 48-well plates at 20,000 cells/well. After 24 h, 5-fold dilutions of the compounds were added. The cells were allowed to proliferate 4 days in the presence of the compounds, trypsinized, and counted by means of a Coulter counter (Analis, Belgium). HeLa and MDA-MB-231 cells were seeded in 48-well plates at 10,000 cells/well. After 24 h, different concentrations of the compounds were added. After 3 days of incubation, the cells were trypsinized and counted in a Coulter counter. Suspension cells (human lymphoid Cem cells) were seeded in 96-well plates at 60,000 cells/well in the presence of different concentrations of the compounds, allowed to proliferate for 96 h, and counted in a Coulter counter. The 50% inhibitory concentration (IC₅₀) was defined as the compound concentration required to reduce cell proliferation by 50%.

Bueno O., Estévez Gallego J., Martins S., Prota A.E., Gago F., Gómez-SanJuan A., Camarasa M.J., Barasoain I., Steinmetz M.O., Díaz J.F., Pérez-Pérez M.J., Liekens S, & Priego E.M. (2018). High-affinity ligands of the colchicine domain in tubulin based on a structure-guided design. Scientific Reports, 8, 4242.

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Cell Proliferation Assay Protocol

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HMEC-1 cells were seeded in 48-well plates at 20,000 cells/well. After 24 h, 5-fold dilutions of the compounds were added. The cells were allowed to proliferate 4 days in the presence of the compounds, trypsinized, and counted by means of a Coulter counter (Analis, Belgium). Human cervical carcinoma (HeLa) cells were seeded in 48-well plates at 10,000 cells/well. After 24h, different concentrations of the compounds were added. After 3 days of incubation, the cells were trypsinized and counted in a Coulter counter. Suspension cells (Mouse leukemia L1210 and human lymphoid CEM cells) were seeded in 96-well plates at 60,000 cells/well in the presence of different concentrations of the compounds, allowed to proliferate for 96 h, and counted in a Coulter counter. The 50% inhibitory concentration (IC 50 ) was defined as the compound concentration required to reduce cell proliferation by 50%.

Bueno O., Gargantilla M., Estévez-Gallego J., Martins S., Díaz J.F., Camarasa M.J., Liekens S., Pérez-Pérez M.J, & Priego E.M. (2019). Diphenyl ether derivatives occupy the expanded binding site of cyclohexanedione compounds at the colchicine site in tubulin by movement of the αT5 loop. European journal of medicinal chemistry, 171.

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Cell Size and Count Analysis

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Cell counts and cell size analysis
were performed by Coulter counter using RBC lysed blood and DU145
cancer cells for both live and fixed cells. Whole blood was lysed
and washed as described earlier and resuspended in FACS buffer. DU145
cells were harvested and resuspended in FACS buffer as reported. For
fixing cells, 4% fresh PFA was used. All cells were kept on ice prior
to the Coulter counter analysis. FACS buffer was used for background
measurements. The Coulter counter measures the change in electrical
impedance when a cell passes through an orifice, which is recorded
as a voltage pulse that is proportional to the volume displaced by
the cell.

Undvall Anand E., Magnusson C., Lenshof A., Ceder Y., Lilja H, & Laurell T. (2021). Two-Step Acoustophoresis Separation of Live Tumor Cells from Whole Blood. Analytical Chemistry, 93(51), 17076-17085.

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Proliferation of Pancreatic Cancer Cells

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Cultures of PANC-1 and MiaPaCa-2 cells, 3–5 days after passage, were washed and suspended in DMEM containing 5 mM glucose. Cells were then disaggregated by two passes through a 19-guage needle into an essentially single-cell suspension as judged by microscopy. Cell number was determined using a Coulter Counter, and 2×10⁴ cells were seeded in 35 mm tissue culture plates in DMEM containing 5 mM glucose and 10% FBS. After 24 h of incubation, the medium was removed and the cultures shifted to DMEM containing 5 mM glucose without or with 3% FBS. The cultures were then incubated in a humidified atmosphere containing 10% CO₂ at 37°C for 4 days and the total cell count was determined from a minimum of four dishes per condition using a Coulter Counter, after cell clumps were disaggregated by passing the cell suspension ten times through a 19- and subsequently a 21-gauge needle.

Ming M., Sinnett-Smith J., Wang J., Soares H.P., Young S.H., Eibl G, & Rozengurt E. (2014). Dose-Dependent AMPK-Dependent and Independent Mechanisms of Berberine and Metformin Inhibition of mTORC1, ERK, DNA Synthesis and Proliferation in Pancreatic Cancer Cells. PLoS ONE, 9(12), e114573.

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Evaluating Amino Acid Uptake in Irradiated Tumor Cells

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Harvested and EMEM washed HSG tumor cells were counted using a Coulter counter and the cell concentration was adjusted to 1 x 10⁶ cells or 2 x 10⁶ cells that were plated in T-25 flasks. These flasks were incubated at 37°C in 5% CO₂ for 1 day. The cell bearing flasks were then irradiated with a carbon-ion beam to attain a dose of 3-Gy. Post-irradiation, the media in each flask was changed to fresh media, and flasks were incubated for the appropriate time for subsequent amino acid uptake studies conducted on the assigned day. On the study day, the cells were washed, harvested and counted using a Coulter counter to provide the surviving tumor cell number. The cell concentration then was adjusted to 2 x 10⁶/mL and the cell uptake of ¹⁴C-MeAIB or ¹⁴C-Met by HSG cells was investigated from 1–5 days post-irradiation using the procedure described above.

Uehara T., Watanabe M., Suzuki H., Furusawa Y, & Arano Y. (2017). Amino acid transport system - A substrate predicts the therapeutic effects of particle radiotherapy. PLoS ONE, 12(2), e0173096.

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Quantifying Cell Growth Kinetics

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For growth rate determination, cells were plated in 35-mm dishes at the density of 30 000 cells per dish in their culture media. Starting from day 2, 2 dishes of each line were taken out each day and nuclei were released according to the protocol described previously and counted using a Coulter counter.^{12 (link)} The culture medium was refreshed every other day.
To determine the effect of SKF38393 (a specific D₁R agonist) on cell growth and to compare the sensitivity MCF-7 cells with or without Cav-1 to this compound, cells were plated at a concentration of 30 000 cells per well in 6-well plates. Two days later, the cells were treated with the dopamine type 1 receptor (D₁R) agonist from Smith Kline French drug number 38393 (SKF38393) at various concentrations for 5 days with medium change on day 3. Cell number was counted using a Coulter counter.^{12 (link)}

Chai T., Yue W., Xu P., Gildea J, & Felder R. (2024). Caveolin-1, a Determinant of the Fate of MCF-7 Breast Cancer Cells. Breast Cancer : Basic and Clinical Research, 18, 11782234241226802.

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PBMC Isolation and Immune Cell Quantification

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Peripheral blood mononuclear cells (PBMCs) were isolated from 20 mL of heparinized blood using Ficoll density gradient centrifugation (Abbott Laboratories, Chicago, IL, USA) and cryopreserved until analysis. Total lymphocyte and monocyte counts were determined in fresh ethylenediaminetetraacetic acid (EDTA)-treated blood using a Coulter counter.

Rodero-Romero A., Sainz de la Maza S., Fernández-Velasco J.I., Monreal E., Walo-Delgado P.E., Chico-García J.L., Villarrubia N., Rodríguez-Jorge F., Rodríguez-Ramos R., Masjuan J., Costa-Frossard L, & Villar L.M. (2023). Blood CD8+ Naïve T-Cells Identify MS Patients with High Probability of Optimal Cellular Response to SARS-CoV-2 Vaccine. Vaccines, 11(9), 1399.

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Gelatin Zymography for Metalloproteinase Quantification

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Endothelial (HUVEC) and tumor cells (MDA-MB231 and HT1080) were incubated for 24 h with their respective media without FBS containing 0.1% BSA, 200 KIU/mL aprotinin together with DMSO (for negative control) and several concentrations of danthron (10, 25 and 50 μM). Then, conditioned media were collected, centrifuged at 1000× g for 10 min at 4 °C and used for gelatin zymography as previously described [11 (link)]. Using a Coulter counter, samples were normalized for equal cell number before being subjected to non-reducing SDS/PAGE with gelatin (1 mg/mL) added to the 10% resolving gel. After electrophoresis, gels were washed twice with 50 mM Tris/HCl, pH 7.4 supplemented with 2% Triton X-100, and incubated for 16 h at 37 °C in a substrate buffer (50 mM Tris/HCl, pH 7.4 supplemented with 1% Triton X-100, 5 mM CaCl₂, and 0.02% Na₃N). Finally, gels were stained with Coomassie blue R-250, and photographed with the ChemiDocTM XRS + System (Bio-Rad) using Image LabTM software 6.1. Quantitative analysis was performed using FIJI software.

Vidal I., Torres-Vargas J.A., Sánchez J.M., Trigal M., García-Caballero M., Medina M.Á, & Quesada A.R. (2023). Danthron, an Anthraquinone Isolated from a Marine Fungus, Is a New Inhibitor of Angiogenesis Exhibiting Interesting Antitumor and Antioxidant Properties. Antioxidants, 12(5), 1101.

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Quantifying Neutrophil Influx in LPS-Induced Peritonitis

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Mice were injected intraperitoneally with 1 mg/kg lipopolysaccharide (LPS) from Escherichia coli O127:B8 (Sigma‐Aldrich, Dorset, UK) or vehicle (PBS) in a volume of 8 ml/kg. At 6 hr, peritoneal lavage was performed using 5 ml of lavage buffer (PBS containing 0·1% bovine serum albumin and 1 mm EDTA). Neutrophils in lavage fluid were quantified using Coulter Counter and haemocytometry measurements, combined with flow cytometry (see below).

Giles J.A., Greenhalgh A.D., Denes A., Nieswandt B., Coutts G., McColl B.W, & Allan S.M. (2018). Neutrophil infiltration to the brain is platelet‐dependent, and is reversed by blockade of platelet GPIbα. Immunology, 154(2), 322-328.

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Chondrocyte Expansion and Characterization

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Primary ACs were passaged in monolayers to P2, P3, P4, P5, P6, and P7 in chondrogenic medium + 5 ng/mL bFGF (Peprotech, Rocky Hill, NJ) in a chondrogenically tuned process, as previously described [11 (link)]. Briefly, P0 ACs were thawed and seeded into T225 flasks at ~25,000 cells/cm². For the first 24 h, 10% FBS was supplemented to promote cell adhesion. The medium was changed every 3–4 days. All cultures took place at 37 °C/10% CO₂. When cells reached 95% confluence, the monolayers were cultured for an additional 4 days. Cell sheets were then lifted with 20 min incubation in 0.25% trypsin/EDTA (Invitrogen) and the sheets digested with 500 units/mL collagenase in chondrogenic medium + 3% FBS for 30 min. Cells were filtered through a 70 μm cell strainer, washed three times, counted with a hemocytometer and Coulter counter (Beckman-Dickinson Z-100), and either passaged again or subjected to aggregate culture, as described next. All passaging metrics are presented in Table 1. Cell diameter was determined through analysis with a Coulter counter. The following formulas were used for calculating the cell expansion metrics: cell doubling number = log(expansion factor)/log(2) (i.e., the number of times a cell population has doubled); expansion factor = initial number of cells/final number of cells.

Huang B.J., Hu J.C, & Athanasiou K.A. (2016). Effects of passage number and post-expansion aggregate culture on tissue engineered, self-assembled neocartilage. Acta biomaterialia, 43, 150-159.

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