Cell counting kit-8 (CCK-8) was used to evaluate the cytotoxicity of γ-MnO2 nanomaterials using HFF-1 cells. HFF-1 cells were incubated in DMEM culture medium supplemented with 15% bovine serum solution in a humidified atmosphere of 5% CO2 at 37 °C. 100 μL of HFF-1 cell suspension was added in a 96-well cell-culture plate at a density of 5 × 103 cells per well, and incubated for 24 h to allow attachment. 100 μL microsphere-like γ-MnO2 nanoparticle suspension with four different final concentrations (0.1, 1, 10, and 50 μg mL−1) was then added into the wells. 100 μL DMEM medium was added in other wells as control groups. After the incubation of 24 h, 10 μL CCK-8 solution (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfobenzene)-2H-tetrazole monosodium salt) was added into each well. The 96-well plate was incubated for 2 h (5% CO2 at 37 °C). The optical density at 450 nm was measured by Tecan Infinite F200/M200 Multifunction Microplate Reader (Tecan, Männedorf, Switzerland). Cell viability was calculated as the mean absorption value of four measurements of each condition divided by the mean absorption value of the DMEM control group.
Infinite f200 m200 multifunction microplate reader
Manufactured by Tecan
Sourced in Switzerland
The Infinite F200/M200 is a multifunction microplate reader designed for a variety of absorbance, fluorescence, and luminescence detection applications. It features a monochromator-based optical system and supports various microplate formats. The reader can be used for a range of assays and experiments in life science research and drug discovery.
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3 protocols using infinite f200 m200 multifunction microplate reader
1
Cytotoxicity Evaluation of γ-MnO2 Nanoparticles
2
Evaluating YCI's Effect on Human Fibroblasts
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Human skin fibroblasts (HFF-1) cells were grown in complete cell culture medium (1% FBS, 2% Penicillin–Streptomycin, DMEM medium). The cells were plated at a density of 5000 cells per well in a 96-well culture plates, and incubated in a humidified atmosphere of 5% CO2 at 37 °C for 24 hrs. 0.1 mg/mL YCI in DMEM medium was added to the wells. The cells cultured in DMEM in the absence of YCI was used as control group. The cells were cultured in a humidified atmosphere of 5% CO2 at 37 °C for 24 hrs, 48 hrs, 72 hrs and 96 hrs, respectively. 10 μL of CCK-8 reagent was added into each well, and incubated for 4 hrs at 37 °C in darkness. The optical density at a wavelength of 450 nm was measured using a Tecan Infinite F200/M200 multifunction microplate reader (Tecan, Männedorf, Switzerland). Cell viability was calculated as the mean absorption value of the YCI-treated group divided by the mean absorption value of the control group.
3
Evaluating Cell Viability of YCI Treatment
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HeLa cells were grown to a cell density of 1 × 105 cells/mL in complete cell culture medium (1% FBS, 2% Penicillin–Streptomycin, DMEM medium) in a humidified atmosphere of 5% CO2 at 37 °C. The cells were plated at a density of 5000 cells per well in a 96-well culture plates, and incubated for 24 hrs to allow for attachment. The cells were treated with different concentrations of YCI (0.01, 0.05, 0.1, 0.5, 1 mg/mL) for 24 hrs. The cells cultured in DMEM in the absence of YCI was used as control group. 10 μL of CCK-8 reagent was added into each well, and incubated for 4 hrs at 37 °C in darkness. The optical density at a wavelength of 450 nm was measured using a Tecan Infinite F200/M200 multifunction microplate reader (Tecan, Männedorf, Switzerland). Cell viability was determined as the mean absorption value of the YCI-treated group divided by the mean absorption value of the control group.
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