Cells in 12-well plates at 4×104 cells per well were cultured overnight in regular media and then placed in conditioned media (diluted 1:1 with fresh media) collected from day-6 or day-9 culture of irradiated or non-irradiated cells. At 72-hour post-culture, viable cells were counted using an automated Countess® Cell Counter (Invitrogen, CA). Average values from triplicate wells were calculated. For pathway activation assay, cells at 72 hours post-culture were analyzed by Western blotting for levels of AKT/phospho-AKT, ERK/phospho-ERK and STAT5/phospho-STAT5. Western blot assay was performed on two biological replicates.
Countess cell counter
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Countess cell counter is a compact and automated instrument designed for cell counting and viability analysis. It utilizes advanced optics and digital image processing to rapidly determine the number and health of cells in a sample. The Countess provides consistent and reliable cell count data, supporting various applications in cell biology and biomedical research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (9 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Tryple express, by Thermo Fisher Scientific (5 mentions)
Trypsin edta solution, by Merck Group (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Novaseq, by Illumina (3 mentions)
Trypan blue, by Thermo Fisher Scientific (3 mentions)
Axiovert 40 cfl, by Zeiss (3 mentions)
Moticam 2300 camera, by Motic (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (2 mentions)
Colo205 ccl 222 tumor cells, by American Type Culture Collection (2 mentions)
Rpmi 1640 cell culture medium, by American Type Culture Collection (2 mentions)
Trustain fcx, by BioLegend (2 mentions)
Ampure xp bead, by Beckman Coulter (2 mentions)
Bioanalyzer, by Agilent Technologies (2 mentions)
Sh800 cell sorter, by Sony (2 mentions)
Glutamine, by Thermo Fisher Scientific (2 mentions)
138 protocols using countess cell counter
1
Conditioned Media Effects on Cell Viability
2
Culturing Pancreatic Cancer Cell Lines
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The human pancreatic cancer cell lines MIA PaCa-2, AsPC-1, PANC-1 and BxPC-3 were purchased from American Type Culture Collection (CRL-1420TM, CRL-1682, CRL-1469 and CRL-1687, respectively, ATCC; Rockville, MD, USA) and banked at Centre Paul Strauss. Cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2 and 95% air. PANC-1 was cultured in Dulbecco’s modified Eagle’s medium (DMEM; PAN Biotech GmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS, PAN Biotech GmbH) and 1% of a solution of penicillin (10000 IU/mL) and streptomycin (10 mg/mL) (PAN Biotech GmbH). MIA PaCa-2 was cultured in the same conditions with 1% of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, 1 mM, PAN Biotech GmbH), 1% of sodium pyruvate (10 mM, PAN Biotech GmbH) and 1% of non-essential amino acids (NEAA, PAN Biotech GmbH). AsPC-1 and BxPC-3 were cultured in Roswell Park Memorial Institute medium (RPMI; PAN Biotech GmbH) supplemented with 10% FBS, and 1% of a solution of penicillin (10000 IU/mL) and streptomycin (10 mg/mL). Subconfluent cell monolayers were trypsinized once a week using 0.5% trypsin containing 2% EDTA (PAN Biotech GmbH) and plated at passage ratios between 0.25:10 to 1:10, according to the cell line or used directly for study after enumeration determined with a Countess® Cell Counter (Countess, Invitrogen, Carlsbad, CA, USA).
3
Cell Viability Measurement Using Trypan Blue
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Each cell line was plated in a 6-well dish at a density of 105 cells per well. At the time point indicated cells were harvested and mixed with an equivalent volume of 0.4% Trypan blue. Cell counts and viability was measured using a countess cell counter (Invitrogen).
4
Colorectal Adenocarcinoma COLO205 2D Culture
5
Evaluating Porphyrin Biocompatibility and Phage Efficacy
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H441 cells were seeded into 6-well plates at a concentration of 8.4 × 105 cells/mL and incubated overnight to allow cells to adhere. The next morning, cells were washed and replenished with fresh media, without antibiotics or serum, including the denoted experimental conditions. For ZnPor biocompatibility assessment, the experimental porphyrin was added to the cells at a concentration of 4 or 65 µg/mL. For phage evaluation, H441 cultures were exposed to PEV2 at a concentration that would represent a 1:1 MOI (PsA: phage) in the infection model. All experimental conditions were incubated for 24 h, after which the cells were washed, removed from the cell culture plate via trypsin, and counted using an Invitrogen Countess Cell Counter. Cell viability was determined by comparing experimental conditions against an untreated H441 control. Data is included from three independent experiments.
6
Single-Cell Targeted Sequencing of Hematologic Malignancies
7
Evaluating CCoV Infection Effects on EV
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After cell densities reached 70–80% confluency, the viability of the cells was examined via trypan blue assay. The cells were trypsinized, centrifuged, resuspended, and counted using trypsin, 0.4% trypan blue solution, and a Countess cell counter (Invitrogen, Waltham, MA, USA). CRFK cells were seeded at a density of 5 × 105 cells in each cell culture dish and incubated overnight at 37 °C in a 5% CO2 incubator before infection. Overnight cell-free media were discarded and 3 mL of 2% exosome-free medium was added to each dish for infection experiments. Uninfected control dishes were incubated as they were after adding exosome-free media, whereas infection dishes were infected with CCoV at MOI of 400 infectious units (IFU). Plates were incubated for 48 h and 72 h in independent experiments. The cell-free medium from each control and infection dish was collected independently post-incubation and stored at −80 °C for further EV isolation.
8
GICs/NSC-2201 Cell Growth Kinetics
9
Flow Cytometric Analysis of Cell Lines
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The OS cell lines were harvested and cell numbers were determined by utilizing the Countess Cell Counter (Invitrogen) followed by dilution in flow buffer (Dulbecco’s PBS with 0.5% BSA and 0.1% NaN3). The cell suspension was distributed in a 96 well plate with 2 × 105 cells per sample well. Primary antibodies were added in a concentration of 10 μg/ml and cells were incubated at 4°C for 30 min before three washes in 200 μl flow buffer. Secondary antibody, anti-mouse IgG F(ab’)2 fragment-FITC (Sigma-Aldrich, St Louis, MO, USA) or anti-human IgG-FITC (US Biologicals, Salem, MA, USA), was added and incubated for 30 min and washed as in the previous step. All wash steps were performed by centrifugation at 1,200 rpm for 5 min.
Washed cell pellets were dissolved in flow buffer and analyzed in a FACS Calibur (BD Bioscience, Franklin Lakes, NJ, USA) or Guava EasyCyte HT flow cytometer (Merck Millipore, Darmstadt, Germany). The flow analysis was replicated and reproduced at least three times for all cell lines. The data were analyzed with Kaluza Analysis 1.3 software (Beckman Coulter, Brea, CA, USA).
Washed cell pellets were dissolved in flow buffer and analyzed in a FACS Calibur (BD Bioscience, Franklin Lakes, NJ, USA) or Guava EasyCyte HT flow cytometer (Merck Millipore, Darmstadt, Germany). The flow analysis was replicated and reproduced at least three times for all cell lines. The data were analyzed with Kaluza Analysis 1.3 software (Beckman Coulter, Brea, CA, USA).
10
HPV-18E2 Protein Immunoprecipitation
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U2OS cells were transfected with 1 µg of the pQMN-18E2, pQMCF-1-18E8E2, pQMCF-1-18E2C1, and pQMCF-1-18E2C2 expression vectors. At 36 h hours post-transfection, the cells were detached with PBS containing 3 mM EDTA and counted with a Countess Cell Counter (Invitrogen). The cells were then collected by centrifugation (200 g for 5 minutes), resuspended in RIPA buffer containing 1X protease inhibitor cocktail (Roche) and sonicated. The lysate was incubated with 30 µl of streptavidin-coated magnetic beads (Dynabeads M250) for 2 hours. The beads were collected, and each lysate was transferred to a new tube with 2 µg of biotinylated chicken HPV-18E2C antibody that had been raised against the C-terminal domain of the HPV18 E2 protein and incubated overnight at 4°C end-over-end. The next day, 50 µl of streptavidin-coated magnetic beads were blocked in 3% BSA-PBS for 2 hours, washed with PBS, added to the lysate and incubated for 4 hours at 4°C. The beads were then collected and washed 3 times in RIPA buffer containing 1x protease inhibitor cocktail (Roche). A total of 10 µl of 1x Laemmli buffer with 1 mM DTT was added per 3*10ˇ6 cells. Western blotting was performed as described in [24] (link).
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