CM-H2DCFDA (DCF-DA), which functions as a ROSsensitive fluorophore, was used to detect intracellular ROS generation. Mouse hepatocytes on 35-mm cell culture dishes (BD Bioscience) were treated with H2O2 with or without pretreatment with BHA or NAC for 30 min and cultured at 37℃ for 2 h. The cells were moved into the dark and incubated with 5 µM DCF-DA for 30 min at 37℃. Next, the cells were washed three times with PBS and viewed using fluorescence microscopy (DM IL LED Fluo; Leica, Germany) at 100 × magnification. Fluorescence intensity was measured using Tecan Infinite M200 Pro (Life Technologies).
35 mm cell culture dishes
Manufactured by BD
Sourced in United States
The 35 mm cell culture dishes are circular, flat-bottomed containers made of polystyrene material, primarily used for the in vitro cultivation and maintenance of cells.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using 35 mm cell culture dishes
1
Measurement of Intracellular ROS in Hepatocytes
2
Isolation of Hippocampal Neurons from CA1 Region
3
Quantifying Cell Viability and Apoptosis
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Cell viability assays and apoptosis detection by annexin V/propidium iodide assay.
To determine cell growth inhibition, early and late apoptosis, as well as cell death, the annexin V/propidium iodide assay (ImmunoStep, S.L., Salamanca, Spain) was applied. A total of 200,000 cells were seeded in 35-mm cell culture dishes (Falcon, Corning, NY, USA) and allowed to settle for 24 h. Then, serial dilutions of the chemical compounds (10, 1, 0.1 and 0.01 μM) were added to expose the cells for 24 h. After the incubation period, cells were detached by trypsinization and collected into flow cytometry tubes together with the supernatants. After three washing steps with PBS (1200 rpm, 3 min), 1X annexin-binding buffer was added to re-suspend the cells at a 1 × 106 cells/mL concentration. Then, cells were systematically stained with 5 μL of the annexin-V fluorescein isothiocyanate (FITC) and 5 μL of the propidium iodide and incubated for 15 min at room temperature in the dark. After this time, 400 μL of 1X annexin-binding buffer was added. For data analysis of the annexin+/− and PI+/− ratios, the Infinicyt™ software (Cytognos SL, Salamanca, Spain) was used. By representing viability (% annexin− cells) on the vertical and concentrations of the compound on the horizontal one, curves of viability were obtained.
To determine cell growth inhibition, early and late apoptosis, as well as cell death, the annexin V/propidium iodide assay (ImmunoStep, S.L., Salamanca, Spain) was applied. A total of 200,000 cells were seeded in 35-mm cell culture dishes (Falcon, Corning, NY, USA) and allowed to settle for 24 h. Then, serial dilutions of the chemical compounds (10, 1, 0.1 and 0.01 μM) were added to expose the cells for 24 h. After the incubation period, cells were detached by trypsinization and collected into flow cytometry tubes together with the supernatants. After three washing steps with PBS (1200 rpm, 3 min), 1X annexin-binding buffer was added to re-suspend the cells at a 1 × 106 cells/mL concentration. Then, cells were systematically stained with 5 μL of the annexin-V fluorescein isothiocyanate (FITC) and 5 μL of the propidium iodide and incubated for 15 min at room temperature in the dark. After this time, 400 μL of 1X annexin-binding buffer was added. For data analysis of the annexin+/− and PI+/− ratios, the Infinicyt™ software (Cytognos SL, Salamanca, Spain) was used. By representing viability (% annexin− cells) on the vertical and concentrations of the compound on the horizontal one, curves of viability were obtained.
4
Cellular Uptake of BSA-Alexa Fluor 488
5
hiPSCs Transfection with Negative Control
6
R-Phycoerythrin Electroporation Protocol
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