Countess counter

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Countess Counter is a compact and automated cell counting device designed for accurate cell enumeration. It utilizes advanced optics and imaging technology to rapidly count and analyze cell samples. The Countess Counter provides reliable and consistent cell counts, making it a valuable tool for researchers and scientists working in various laboratory settings.

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Lab products found in correlation

Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Canto 2, by BD (2 mentions) Celltiter glo assay kit, by Promega (1 mentions) Tryple reagent, by Thermo Fisher Scientific (1 mentions) Trypan blue staining method, by Thermo Fisher Scientific (1 mentions) Synvisc, by Sanofi (1 mentions) Fixation and permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Facscanto, by BD (1 mentions) Poly d lysine coated plates, by BD (1 mentions) 1480 wizard automatic γ counter, by PerkinElmer (1 mentions)

Close spelling variants of this product

Countess C10281 automated cell counter Countess Automated Cell Counter Hemocytometer C10227 Invitrogen Countess™ II FL automated cell counter Countess II FL automated counter Countess Automated Cell Counter Countess II Automatic Cell Counter Countess 3 Automated Cell Counter Countess Automated Cell Counter system Countess I FL Automated Cell Counter Countess automated counter

15 protocols using countess counter

VSMC Proliferation Assay with PIO

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Subconfluent VSMCs were serum-deprived for 48 hr and then pretreated with PIO (3, 10 or 30 μM, 30 min) or vehicle control followed by exposure to PDGF (30 ng/ml, 96 hr). We replaced the medium with fresh serum-free medium containing the indicated concentrations of PIO and/or PDGF every 48 hr. VSMCs were then trypsinized and the changes in cell number were determined using Countess Counter (Life Technologies), as described [40 (link)].

Osman I, & Segar L. (2015). Pioglitazone, a PPARγ agonist, attenuates PDGF-induced vascular smooth muscle cell proliferation through AMPK-dependent and AMPK-independent inhibition of mTOR/p70S6K and ERK signaling. Biochemical pharmacology, 101, 54-70.

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Modulation of VSMC Proliferation by SFN

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VSMCs were equally seeded onto petri dishes (~4 × 10⁵ cells/60 mm dish) and maintained in culture for 48 h. After the attainment of uniform subconfluency in all dishes, VSMCs were deprived of serum for 48 h. Subsequently, VSMCs were treated with SFN (5 μM) or vehicle control (DMSO, 0.05%) for 30 min followed by exposure to PDGF (30 ng.mL⁻¹) for 96 h. At the 48 h time point, the respective cells were replenished with fresh media with and without SFN or PDGF. After 96 h, VSMCs were trypsinized and the changes in cell number were determined using Countess Counter (Life Technologies), as described [36 (link)].

Shawky N.M, & Segar L. (2017). Sulforaphane inhibits platelet-derived growth factor-induced vascular smooth muscle cell proliferation by targeting mTOR/p70S6kinase signaling independent of Nrf2 activation. Pharmacological research, 119, 251-264.

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VSMC growth modulation by SFN and leptin

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Serum-deprived VSMCs were treated with SFN (5 µM) or vehicle control (DMSO, 0.05%) for 30 min followed by exposure to leptin (6 nM) for 96 hr. At the 48 hr time point, the respective cells were replenished with fresh media with and without SFN or leptin. After 96 hr, VSMCs were trypsinized and the changes in cell number were determined using Countess Counter (Life Technologies), as described [37 (link)].

Shawky N.M., Pichavaram P., Shehatou G.S., Suddek G.M., Gameil N.M., Jun J.Y, & Segar L. (2016). Sulforaphane improves dysregulated metabolic profile and inhibits leptin-induced VSMC proliferation: implications toward suppression of neointima formation after arterial injury in western diet-fed obese mice. The Journal of nutritional biochemistry, 32, 73-84.

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Cell Proliferation Assay for VSMCs

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Subconfluent VSMCs were serum-deprived for 48 h and then treated with D-fructose (5.5 or 25 mM), insulin (100 nM) or PDGF (30 ng/ml) for 96 h. Fresh serum-free media containing the respective treatments were replaced every 48 h. VSMCs were then trypsinized and the changes in cell number were determined using Countess Counter (Life Technologies, Carlsbad, CA), as described (Pyla et al., 2013 (link)).

Osman I., Poulose N., Ganapathy V, & Segar L. (2016). High fructose-mediated attenuation of insulin receptor signaling does not affect PDGF-induced proliferative signaling in vascular smooth muscle cells. European journal of pharmacology, 791, 703-710.

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Cell Viability Assessment by ATP Luminescence

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Cell growth was quantified by counting cells using the Countess Counter (Life Technologies). Cell viability was assessed by chemiluminescence measurement of intracellular ATP using the CellTiter-Glo assay kit (Promega, Madison, WI) as per the manufacturer's instructions. The effects of hit shRNAs and siRNAs were compared to those of a control selected at random from the library (after construction but prior to screening, which is equivalent to a randomly selected sequence from a pool of random sequences made in an oligonucleotide synthesizer).

Cotticelli M.G., Acquaviva F., Xia S., Kaur A., Wang Y, & Wilson R.B. (2015). Phenotypic Screening for Friedreich Ataxia Using Random shRNA Selection. Journal of biomolecular screening, 20(9).

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Hyaluronic Acid Effects on Adipose-Derived Mesenchymal Stem Cells

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In step eight, MSCs were divided into seven groups, which were exposed to hyaluronic acid from six commercial brands and phosphate-saline buffer – PBS (control group):

Group 1: AD-MSC in contact with Fermathron® (Hyaltech),

Group 2 AD-MSC in contact with Orthovisc® (J&J),

Group 3 AD-MSC in contact with Synvisc® (Sanofi),

Group 4 AD-MSC in contact with Synovium® (LCA Pharmaceutical),

Group 5 AD-MSC in contact with Suprahyal® (Zodiac),

Group 6 AD-MSC in contact with Osteonil® (TRB Chemedica),

Group 7 AD-MSC in contact with PBS (Thermo Fisher Scientific).

In the preparation process, the cells were detached from the culture bottle with TrypLE reagent (Thermo Scientific) and centrifuged at 300 × g for 5 min, counted in the Countess apparatus (Thermo Fisher), and resuspended in hyaluronic acid or PBS at the concentration of. After 4, 24, and 48 h of incubation at room temperature and atmosphere, cell viability analysis was performed through the Countess counter by the Trypan Blue staining method (Thermo Fisher Scientific), according to the supplier's recommendations. Two independent experiments were performed, each with three different AD-MSC strains (AT8, AT11, and AT13). For each time interval, a single viability reading was made, and the same experimental conditions were followed.

Kaleka C.C., Zucconi E., Vieira T.D., Secco M., Ferretti M, & Cohen M. (2018). Evaluation of different commercial hyaluronic acids as a vehicle for injection of human adipose-derived mesenchymal stem cells. Revista Brasileira de Ortopedia, 53(5), 557-563.

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Trypan Blue Assay for Lung Cancer Cell Viability

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Cell viabilities were determined using trypan blue exclusion assay. To estimate viability changes, 1×10⁴ human lung carcinoma cells were added with various concentrations of aqueous BJ extract and incubated at 37°C for 12 hours, while water alone served as control group. Cells as collected by trypsin–ethylenediaminetetraacetic acid were stained with trypan blue. Cells with exclusion of the dye were counted as viable and measured with a Countess™ counter (Thermo Fisher Scientific). The viability was determined by comparing cell numbers of treated cells with those of water control as percentages. The inhibition concentration, IC₅₀, defined as concentration that inhibited 50% of cell proliferation, was calculated from a calibration curve by linear regression using Microsoft Excel.

Kim S.H., Liu C.Y., Fan P.W., Hsieh C.H., Lin H.Y., Lee M.C, & Fang K. (2016). The aqueous extract of Brucea javanica suppresses cell growth and alleviates tumorigenesis of human lung cancer cells by targeting mutated epidermal growth factor receptor. Drug Design, Development and Therapy, 10, 3599-3609.

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Evaluating WHCO1 Cell Growth on Decellularized ECMs

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Cell growth curves or proliferation rates after plating WHCO1 on decellularised ECMs and/or treatment with cisplatin, 5-fluro-uracil and epirubicin drugs were determined using the Countess Counter (Thermo Fisher Scientific, Waltham, MA, USA). WHCO1 cells (5 × 10⁵) were plated on the decellularised ECMs and/or treated with the drugs for the indicated time periods. Cells were trypsinised and centrifuged at 1800 rpm for 5 min. Cells were suspended in 2 mL complete media and 10 µL was mixed with Trypan Blue. In addition, WHCO1 cells were plated in 96-well plates with or without the ECMs and allowed to grow for about 48–72 h in either drug-free medium or under treatment with increasing concentrations of cisplatin, 5-fluro-uracil and epirubicin drugs. The IC₅₀ for each cell population was measured using the MTT assay. Briefly, WHCO1 cells were plated in 96-well plates with or without ECMs overnight. Drugs were added and incubation continued for 24 h. The MTT reagent was added and the cells were shaken. Colour changes were read on a microplate reader.
WHCO1 cancer cell doubling time (PD) is the time it takes the population of WHCO1 cells to double and was calculated based on the following formula:
where t equals time in hours, ln represents the natural logarithm, FCC represents the final WHCO1 cancer cell number, and SCC represents the starting WHCO1 cancer cell number.

Senthebane D.A., Jonker T., Rowe A., Thomford N.E., Munro D., Dandara C., Wonkam A., Govender D., Calder B., Soares N.C., Blackburn J.M., Parker M.I, & Dzobo K. (2018). The Role of Tumor Microenvironment in Chemoresistance: 3D Extracellular Matrices as Accomplices. International Journal of Molecular Sciences, 19(10), 2861.

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Transient Transfection of COS7 Cells

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Culture of Cos7 cells was performed as previously described (6 (link),8 (link)). Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 8% heat inactivated FBS and kept in a 37°C humidified 5% CO₂ incubator. Cells were seeded into 96-well plates at a density of 15,000–20,000 cells per well (determined using a Countess Counter^™, Invitrogen) one day prior to transfection. Cells were transiently transfected using polyethylenimine as described previously (6 (link),8 (link)). Hemagglutinin-tagged human CLR, hemagglutinin-tagged human CTR (CT_(a) subtype) and myc-tagged human RAMP1 in pcDNA3 or 3.1 plasmid vectors were under the control of the human cytomegalovirus promotor and used as previously described (6 (link),7 (link)).

Walker C.S., Raddant A.C., Woolley M.J., Russo A.F, & Hay D.L. (2017). CGRP receptor antagonist activity of olcegepant depends on the signalling pathway measured. Cephalalgia : an international journal of headache, 38(3), 437-451.

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Characterization of Isolated T Cells

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Isolated T cells were collected by centrifugation (330 × g, 5 min at 4 ℃) and cell counts were determined by using a Countess Counter (Invitrogen). For surface staining, cells were resuspended in PBS (with 2% FBS) containing the antibody, incubated on ice for 30 min, washed in PBS (with 2% FBS), and analyzed using flow cytometry. Flow cytometry assay was performed on CANTO II (BD Biosciences). For intracellular molecule detection, fixation and permeabilization buffers (eBioscience) were used according to the instructions. Data were collected on BD CANTO II (BD Biosciences) with FACSCanto software version 2.1 (BD). Antibodies used were described in Table S2.

Wang X., Shen H., Zhangyuan G., Huang R., Zhang W., He Q., Jin K., Zhuo H., Zhang Z., Wang J., Sun B, & Lu X. (2018). 14-3-3ζ delivered by hepatocellular carcinoma-derived exosomes impaired anti-tumor function of tumor-infiltrating T lymphocytes. Cell Death & Disease, 9(2), 159.

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