Apoptosis was determined by a CellEvent™ Caspase-3/7 green ReadyProbes™ reagent (Life Technologies, Grand Island, NY, USA). Briefly, A549 cells were seeded at a density of 100,000 cells/chamber in 2-well glass chamber slides (SPL Life Sciences, Pocheon-si, Korea) and were treated with 50 μg/mL and 200 μg/mL Fe3O4@SiO2-SH nanoparticles for 24 h. Cells treated with 10 μM cisplatin were used as a positive control and cells treated with H2O were used as a negative control. After treatment, 2 drops/mL of the CellEvent™ Caspase-3/7 green ReadyProbes™ reagent were added to the treated cells and the slides were incubated for 30 min in a humidified incubator at 37 °C and 5% CO2. Apoptotic cells with activated caspase-3/7 show bright green nuclei, while cells without activated caspase 3/7 exhibit minimal fluorescence signal. These cells were observed using a Nikon epifluorescence microscope system Eclipse 80i; the exposure time and dynamic range of the camera in all channels were adjusted to the same values for all of the slides to portray quantitatively comparable images. Images were further analyzed using NIS-Elements Advanced Research 5.11 (instruments and software from Nikon, Tokyo, Japan).
Epifluorescence microscope system eclipse 80i
Manufactured by Nikon
The Nikon epifluorescence microscope system Eclipse 80i is a laboratory equipment designed for fluorescence imaging. It features a modular design that allows for customization to meet specific research requirements. The system utilizes LED illumination and incorporates a range of optical components to enable high-quality fluorescence observation and image capture.
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2 protocols using epifluorescence microscope system eclipse 80i
1
Apoptosis Assay of Fe3O4@SiO2-SH Nanoparticles
2
Apoptosis Detection using Caspase-3/7 Assay
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Apoptosis was determined by a CellEvent ™ Caspase-3/7 green detection reagent (Life Technologies, Grand Island, NY, USA). Briefly, cells were seeded at a density of 300,000 cells/chamber in 2-well glass chamber slides (Thermo Fisher Scientific, Waltham, MA, USA) and were treated for 24 h with silica coated IONPs 100 μg/mL. HK-2 cells treated with 100 μM cisplatin were used as a positive control. After treatment, the cells were labelled with 5 μL of a CellEvent ™ Caspase-3/7 green detection reagent in a complete medium and incubated for 30 min in a humidified 37 °C, CO 2 incubator (5%) in the dark. Images of all of the examined slides were obtained by a Nikon epi-fluorescence microscope system Eclipse 80i; the exposure time and dynamic range of the camera in all of the channels were adjusted to the same values for all of the slides to portray quantitatively comparable images. Images were further processed and merged using NIS-Elements Advanced Research 4.13 (instruments and software from Nikon, Tokyo, Japan).
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