At 6-well plates, MIA PaCa-2 and PANC-1 cells (5 × 105 cells/well) were pretreated with 200 or 400 μg/mL TiOxNPs for one hour followed by exposure to 2 or 5 Gy radiation treatment, and incubation at 37 °C for 48 h. Living cells were stained with trypan blue and counted after 24 and 48 h using a Countess II automated cell counter (Invitrogen Life Technologies, USA). For spheres, cells were first dissociated, and single cell suspensions were obtained, where 2 × 104 cells were treated with 200 or 400 μg/mL TiOxNPs for one hour followed by exposure to 2 or 5 Gy radiation doses. Cells were plated in 96-wells plate in serum-free medium containing sphere-forming growth factors. Living cells were stained with trypan blue and counted after 24 and 48 h using a Countess II automated cell counter (Invitrogen Life Technologies).
Countess 2 automated cell counter
Manufactured by Thermo Fisher Scientific
Sourced in United States, China, Germany, Italy
The Countess II Automated Cell Counter is a compact and easy-to-use device designed for accurate cell counting and viability analysis. It utilizes advanced optics and image processing technology to provide reliable cell counts and viability percentages. The Countess II is a self-contained instrument that requires minimal user interaction, making it suitable for a wide range of applications in cell biology research and cell-based assays.
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Lab products found in correlation
Trypan blue, by Thermo Fisher Scientific (23 mentions)
Trypan blue solution, by Thermo Fisher Scientific (11 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Hiseq 4000, by Illumina (6 mentions)
Trypsin edta, by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Penicillin, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Trypan blue reagent, by Thermo Fisher Scientific (5 mentions)
Tryple, by Thermo Fisher Scientific (5 mentions)
Chromium controller, by 10x Genomics (4 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Ficoll paque plus, by GE Healthcare (4 mentions)
Dnase 1, by Merck Group (4 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions)
Countess automated cell counter, by Thermo Fisher Scientific (3 mentions)
Celltiter glo reagent, by Promega (3 mentions)
Accutase, by Merck Group (3 mentions)
370 protocols using countess 2 automated cell counter
1
Enhancing Radiation Therapy with TiOxNPs
2
Isolation and Enumeration of Murine Immune Cells
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Spleens were obtained from sacrificed mice, were immediately weighed and subjected to digestion with DNAse (100 µg/ml) and Collagenase A (1 mg/ml) in complete media for 30 min at 37°C. Cells were resuspended following filtration through a 70-µm filter. Red blood cells were lysed, white cells were counted using a Countess II Automated Cell Counter (ThermoFisher, Waltham, MA) and spleen cells were analyzed. Liver leukocytes were obtained following homogenization of livers and filtration through a 70-µm filter. Leukocytes were isolated at the interface of a 35%/75% Percoll solution gradient (GE Healthcare, Chicago, IL). Peritoneal cells were obtained immediately following euthanasia through an abdominal incision. The peritoneum was washed with 5 ml of sterile saline and collected. All cell counts were performed using a Countess II Automated Cell Counter (ThermoFisher, Waltham, MA). A minimum of 2 × 106 events were analyzed for each sample.
3
Assessing Retroviral Transduction Effects
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Cells were infected with PrxII-encoding retroviruses for 48 h. Then the cells were trypsinized and counted using a CountessII Automated Cell Counter (ThermoFisher, Waltham, MA, USA). The cells (3 × 103) were plated in 60 mm culture dishes and grown in fresh media at 37 °C in a humidified incubator with 5% CO2 for 10 days. Then the cells were washed with PBS, fixed with 3.7% paraformaldehyde for 5 min and stained with 0.05% crystal violet for 30 min. After washing the cells with cold PBS twice and air-drying for several minutes, the dishes were photographed. The number of colonies was counted using the ImageJ software.
Cell growth was examined using a WST1 assay (Roche Diagnostics, Indianapolis, IN, USA). Retrovirus-infected cells (3 × 103) were seeded in a 96-well culture plate. After infection for the indicated time periods, 10 μl of WST1 reagent was added, followed by incubation at 37 °C incubator for 1 h. The absorbance was measured at 450 nm using 600 nm as a reference wavelength with an EPOCH12 microplate reader (BioTek, Winooski, VT, USA) and averaged from triplicate wells. Cell viability was monitored by trypan blue staining. Cells were trypsinized, stained with trypan blue and then counted using a CountessII Automated Cell Counter (ThermoFisher).
Cell growth was examined using a WST1 assay (Roche Diagnostics, Indianapolis, IN, USA). Retrovirus-infected cells (3 × 103) were seeded in a 96-well culture plate. After infection for the indicated time periods, 10 μl of WST1 reagent was added, followed by incubation at 37 °C incubator for 1 h. The absorbance was measured at 450 nm using 600 nm as a reference wavelength with an EPOCH12 microplate reader (BioTek, Winooski, VT, USA) and averaged from triplicate wells. Cell viability was monitored by trypan blue staining. Cells were trypsinized, stained with trypan blue and then counted using a CountessII Automated Cell Counter (ThermoFisher).
4
Culturing and Counting DMSCs
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The isolated DMSCs were diluted with DMEM/F12 medium and then inoculated into the 24-hole culture plate at a density of 2×105/ml to be maintained at 37℃ and 5% CO2. Half of the medium was replaced every third day. DMSCs were stained with trypan blue and counted every 3 days using an Invitrogen cell counter (Countess II Automated Cell Counter, Thermo Fisher Scientific).
5
Radiation Effects on RD Cells
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RD cells at 96 h after radiation exposure (4 Gy) and 144 h after siRNA transfection (si-DNMT3A, si-DNMT3B or si-NC) were labelled with trypan blue (Invitrogen) in a 1:1 mixture and counted using the Countess II Automated Cell Counter (Invitrogen), according to the manufacturer’s instructions.
For MTT assays, 24 h before the transient transfection with the specific siRNA against DNMTs or the negative control, RD cells were plated at 1.0 × 104 cells/well in 96-well plates and after additional 48 h were exposed or not to radiation. At different time points after irradiation (0–48–72 h), the cell viability was analysed by using MTT [3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium] assay. Specifically, 10 μL of MTT (5 mg/mL, Sigma-Aldrich) were added to each well and plates were incubated at 37 °C for 3 h. Media were then replaced with 100 μL of DMSO and RD cells were incubated at 37 °C for 10 min. Blank cell-free wells were also included. Absorbance was read at wavelength of 560 nm by using GloMax Discover Microplate reader (Promega, Madison, WI, USA). The results were plotted as means ± SD of two independent experiments each performed in triplicate.
For MTT assays, 24 h before the transient transfection with the specific siRNA against DNMTs or the negative control, RD cells were plated at 1.0 × 104 cells/well in 96-well plates and after additional 48 h were exposed or not to radiation. At different time points after irradiation (0–48–72 h), the cell viability was analysed by using MTT [3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium] assay. Specifically, 10 μL of MTT (5 mg/mL, Sigma-Aldrich) were added to each well and plates were incubated at 37 °C for 3 h. Media were then replaced with 100 μL of DMSO and RD cells were incubated at 37 °C for 10 min. Blank cell-free wells were also included. Absorbance was read at wavelength of 560 nm by using GloMax Discover Microplate reader (Promega, Madison, WI, USA). The results were plotted as means ± SD of two independent experiments each performed in triplicate.
6
THP-1 Monocyte Biocompatibility Assay
7
Isolation and Characterization of Lung and Peritoneal Cells
8
Cell Viability and Colony Formation
9
Combination Treatment of LNCaP-tet-AR Cells
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LNCaP-tet-AR cells were pre-cultured with 5% CSS. Cells were then plated into 12-well plates supplemented with/out 0.25 mg/ml doxycycline. After 1-2d, cells were treated with DHT for 24 hours (h) and then treated with DMSO, olaparib (S1060, Selleckchem), cisplatin (S1166, Selleckchem), or GSK126 (S7061, Selleckchem) for 0-6d. Cells were trypsinized, collected, and counted by countess II automated cell counter (Invitrogen).
10
Isolation and Preservation of PBMCs and Whole Blood for Downstream Analyses
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For isolation of PBMCs, whole blood was collected in 10 mL EDTA tubes (Becton Dickinson, Franklin Lakes, NJ, USA). Within a maximum of four hours after collection, PBMCs were extracted with Ficoll-Paque technique, using LeucoSep tubes (Greiner Bio-One, Kremsmünster, Austria ). After isolation and cell count with the Countess II Automated Cell Counter (Invitrogen, Carlsbad, CA, USA ), PBMCs were snap frozen and stored at -80°C until further use. On average, 4 mL of whole blood was used to store one vial of PBMCs.
Whole blood was collected in Tempus tubes (Applied Biosystems, Foster City, CA, USA), in which 3 mL of whole blood is stabilized with 6 mL of RNA stabilizing reagent, resulting in a total volume of 9 mL. Tubes were stored at -80°C within four hours after collection until further use.
Whole blood was collected in Tempus tubes (Applied Biosystems, Foster City, CA, USA), in which 3 mL of whole blood is stabilized with 6 mL of RNA stabilizing reagent, resulting in a total volume of 9 mL. Tubes were stored at -80°C within four hours after collection until further use.
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