The human colon adenocarcinoma cell line, Caco-2 was purchased from European collection of cell cultures (ECACC No. 286010202), and the cells were counted using a Nucleo Counter (ChemoMetec, Denmark) cell counter. SQV was provided by Roche Discovery (Welwyn Garden City, UK), LPV by Abbott Laboratories (Chicago, USA), EFV by Dupont Bristol Myers Squibb (New Brunswick, NJ, USA), and NVP by Boehringer Ingelheim (Berkshire, UK). Radiolabelled SQV (3H SQV), LPV (3H LPV), EFV (14C EFV) and NVP (3H NVP) were purchased from Moravek Biochemicals (Brea California, USA). 3H IVM was kindly donated by Dr Iain Gardner of Pfizer (Sandwich, Kent, UK). PZQ, CLZ, DMEM, HBSS, FBS, DMSO and Trypsin-EDTA solution were purchased from Sigma Aldrich (Poole, UK). ACN and MeOH were purchased from VWR Laboratory Supplies (Poole, UK) whereas diethyl ether was purchased from Fisher Scientific, (Loughborough, UK). Ultima Gold liquid scintillation cocktail was obtained from Packard (Groningen, Netherlands). All the other chemicals used were of analytical or HPLC grade. Deionised water used to prepare the solutions or mobile phase, and was purified in an Elga DV 25 pure lab option system (Elga, High Wycombe, Bucks, and UK).
Nucleocounter
Manufactured by ChemoMetec
Sourced in Denmark
The NucleoCounter is a compact, automated cell counter designed for accurate and reliable cell counting. It utilizes the principle of fluorescence microscopy to determine the total number of cells in a sample. The NucleoCounter provides a fast and efficient way to obtain cell counts, making it a valuable tool for various applications in cell biology and biomedical research.
Automatically generated - may contain errors
Lab products found in correlation
Nucleocassette, by ChemoMetec (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Diethyl ether, by Thermo Fisher Scientific (3 mentions)
Trypsin edta solution, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Casy cell counter, by ChemoMetec (3 mentions)
Cytospin, by Thermo Fisher Scientific (3 mentions)
Diff quik, by Thermo Fisher Scientific (3 mentions)
Nucleoview nc 3000, by ChemoMetec (3 mentions)
Ca2 and mg2 free pbs, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Nucleoview 3000, by ChemoMetec (2 mentions)
Cytospin, by Hanil (2 mentions)
May gr nwald giemsa, by Merck Group (2 mentions)
Hyperflask, by Corning (2 mentions)
Fetal calf serum (fcs), by Merck Group (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Hanks balanced salt solution (hbss), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
85 protocols using nucleocounter
1
Characterizing Anticancer Drug Interactions in Caco-2 Cells
2
Adipose-Derived Stromal Vascular Fraction Isolation
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The SVF was isolated from the abdominal adipose tissue of each patient. Approximately 100 ml of lipoaspirate collected from each patient was divided into two 50‐ml sterile syringes. The syringes were stored in a sterile box at 2–8°C and immediately transferred to the laboratory. The SVF was isolated using an ADSC Extraction Kit (GeneWorld, Ho Chi Minh City, Vietnam,
http://geneworld.vn ) according to the manufacturer's instructions. Briefly, 100 ml of lipoaspirate was placed in a sterile, disposable 250‐ml conical centrifuge tube (Corning Life Sciences, Tewksbury, MA,
https://www.corning.com ) and washed twice with sterile phosphate‐buffered saline (PBS) by centrifugation at 400g for 5 minutes at room temperature. The adipose tissue was then digested using SuperExtract Solution (GeneWorld) containing collagenase at 37°C, for 30 minutes with agitation at 5‐minute intervals. The suspension was centrifuged again at 800g for 10 minutes, and the SVF was harvested as a pellet. The pellet was washed twice with PBS to remove any residual enzyme, and resuspended in PBS so that the cell quantity and viability could be measured using an automatic cell counter (NucleoCounter; Chemometec, Lillerød, Denmark,
https://chemometec.com ).
3
Generation of Murine Dendritic Cells
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Mouse bone marrow cells were harvested by flushing the marrow cavities of the femur and tibia bones of male mice with medium under aseptic conditions. The harvested marrow was depleted of erythrocytes and cultured only in complete RPMI-10% FBS. On day 2, the cultured medium was changed with RPMI-10% FBS, 100 ng/mL granulocyte monocyte colony-stimulating factor (GM-CSF), and 50 ng/mL IL-4 (Santa Cruz Biotechnology Inc., Dallas, TX, USA). Half of the medium was replaced with fresh medium containing the same cocktail of cytokines every 2 days. On day 8, antigens were added into the cultured medium at a concentration of 500 ng/mL. On day 10, nonadherent fractions of DCs were harvested. Adherent fractions of DCs were also harvested by incubating with 0.25% Trypsin/EDTA (Sigma-Aldrich). Then, the numbers of harvested DCs were determined by automatic counting using a Nucleocounter® (ChemoMetec A/S, Allerod, Denmark).
Mature DCs generated from mouse bone marrow were directly stained using fluorescent-conjugated monoclonal antibodies, including anti-CD40, CD80, CD83, and CD86, for 30 minutes at 4°C in the dark. Cells were then analyzed using FACSCalibur flow cytometry (BD Biosciences, San Jose, CA, USA) with CellQuest software.
Mature DCs generated from mouse bone marrow were directly stained using fluorescent-conjugated monoclonal antibodies, including anti-CD40, CD80, CD83, and CD86, for 30 minutes at 4°C in the dark. Cells were then analyzed using FACSCalibur flow cytometry (BD Biosciences, San Jose, CA, USA) with CellQuest software.
4
Determining Cell Counts and Growth Rates
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Example 3
Cell counts from suspension cells or immobilized cells were determined either by counting with a CASY® cell counter as described by Schärfe et al., (Biotechnologie in LaborPraxis 10: 1096-1103 (1988)) or by citric acid extraction and fluorescent staining of the nuclei followed by counting with a NucleoCounter® (Chemometec, DK). The specific growth rate (μ) is calculated from the increase of the cell densities (X1) and/or the dilution rate (D) of the steady state of chemostat cultures of suspensions cells over a certain time interval (t): μ=D+In (Xt I X0)/t
5
Cell Counting and Growth Rate Calculation
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Example 2
Cell counts from suspension cells or immobilized cells were determined either by counting with a CASY® cell counter as described by Scharfe et al., Biotechnologie in LaborPraxis 10: 1096-1103 (1988), or by citric acid extraction and fluorescent staining of the nuclei followed by counting with a NucleoCounter® (Chemometec, DK). The specific growth rate (p) is calculated from the increase of the cell densities(Xt) and/or the dilution rate (D) of the steady state of chemostat cultures of suspensions cells over a certain time interval (t):
μ=D+ln(Xt/XO)/t
6
Isolation of Adipose-Derived Mesenchymal Stem Cells
7
Adipose-Derived Regenerative Cell Isolation
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Under general anesthesia, liposuction of 200 mL of adipose tissue was applied along the abdominal wall after making a single periumbilical incision. ADRCs were isolated from the suctioned adipose tissue by the Celution system. The Celution cell-processing device extracts and concentrates the mononuclear fraction of adipose tissue automatically and aseptically, and removes matrix fragments and unwarranted or deleterious cells. It required approximately 1 h to process 250 mL of liposuction tissue. The concentrated cell output was counted using a NucleoCounter (Chemometec, Allerød, Denmark). Finally, we obtained a 5-mL solution containing concentrated ADRCs.12 (link)
8
Fibroblast-Seeded Collagen Matrix Formation
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Fibroblasts were harvested by trypsinization and counted automatically with the NucleoCounter (Chemometec). After centrifugation at 1200 rpm for 5 min, the cells were resuspended in culture medium to a final concentration of 1 x 106 cells/ml. All ingredients for the collagen matrix were stored on ice before the construction was started and mixed in the exact order listed in the following example for the assembly of at least 6 layers: 1040 μl 10x DMEM (Biochrom), 208 μl 1 M NaOH (Merck), 4,4 ml dH2O (ChemSolute), 260 μl 7.5% NaHCO3 (Carl Roth), 2.6 ml fibroblast growth medium, 4.42 ml (9 mg/ml) rat tail collagen I (Corning; final gel concentration of 2.25 mg/ml). The pH of the solution was adapted to 7.4 with 1 M NaOH and 3.25 ml cell suspension (with a final concentration of 5 x 105 cells/layer) was finally added. After gentle trituration, 2.5 ml solution per layer was transferred to a 6 well culture plate. The collagen polymerized by incubation at 37°C in a cell culture incubator, containing 10% CO2 for 30 min. Finally, the gels were detached from the sides of the well to enable the contraction of the matrix within 8 days. The media were changed daily.
9
Impact of Quantum Dots on Human EpiSC Proliferation
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To analyze the impact of QDs on proliferation of human EpiSCs the growth of labeled and unlabeled cells was observed over a period of 10 days. Cells were seeded in triplicates into 6-well culture plates in a density of 4400 cells/cm². One day after seeding, cells were labeled with 10 nM QDs. Unlabeled cells were kept in culture as a control. During the following 10 days the cells were harvested and counted with NucleoCounter (Chemometec, Denmark) at several time points (1, 2, 4, 6, 8 and 10 days). This experiment was done with cells from 3 different patients.
10
Quantifying Cell Proliferation Kinetics
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Cells were plated at 10,000 cells per well in 24‐well culture plates (day 0) in triplicate. Culture medium was replaced daily during the course of the assay. Each cell line was collected by trypsinization at 24‐h increments. Cell concentrations were determined in Via‐1 cassettes using a Nucleocounter with Nucleoview‐3000 software (Chemometec). Cell number was calculated and plotted using Microsoft Excel software.
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