Quantitative assessment of necrotic and apoptotic cells in HepG2 and HT144 cells treated with 1 μg ml−1 of drug loaded CFO@BTO NRs with and without MF assistance (4 mT, 20 minutes) was done using AOPI staining.10 Cells (>90% viability, 1 × 105 cells per ml) were exposed to NRs + DOX and NRs + MTX for 4 hours. NRs-PMA (1 μg ml−1), NTC, free DOX and MTX (5 μM each) were included as controls. After treatment, cells were washed with PBS, stained with AOPI solution (100 : 32 μg ml−1) and observed under fluorescent microscope (Nikon, MicroPhot-SA). Viable cells exhibit green, necrotic cells exhibit red and apoptotic cells show yellow to orange fluorescence. Quantitative analysis of viable, apoptotic, and necrotic cells was performed using Optika Pro View (Version: x86, 3.7.13977.20190224) software and compared to NTC.
Pro view
Manufactured by Optika
Sourced in Italy
The Pro View is a high-quality laboratory equipment designed for precise visual analysis. It features a durable construction, advanced optics, and an intuitive control interface. The core function of the Pro View is to provide clear and detailed visual inspection of samples under controlled conditions.
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Lab products found in correlation
4 protocols using pro view
1
Quantitative Analysis of Cell Death in HepG2 and HT144 Cells
2
Apoptosis and Necrosis Quantification in Treated Cells
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In order to determine the extent of apoptosis and necrosis in treated cells, AOPI staining was used as previously described [30 (link)]. Pre-seeded HepG2 and HT144 cells (>90% viability, 1.0 × 105 cells/mL) were treated with drug-loaded NPs (5 and 10 µg/mL) for 3 h under appropriate culture conditions. Control groups included NTC, NPs-PMA (10 μg/mL), free DOX and free MTX (5 μM each). Afterwards, the cells were washed with 1× PBS and stained with AOPI (100 and 32 μg/mL) for 1 min at room temperature and visualized under a fluorescence microscope (200×, Nikon, MicroPhot-SA). Green fluorescence indicates viable cells, red fluorescence indicates necrotic cells, whereas yellow to orange fluorescence indicates early and late apoptotic cells, respectively. By using an Optika Pro View (version x86, 3.7.13977.20190224) instrument, live, necrotic, and apoptotic cells were counted, and their percentages were calculated relative to NTC.
3
Microscopy Characterization of Microspheres and HPM Composites
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Microspheres and HPM composites were characterized with an IM-5FLD inverted fluorescence microscope (Optika Srl, Ponteranica, Italy) equipped with (i) an 8W XLED illumination source for sample analysis under transmitted light; (ii) 5W LED excitation illumination sources at 470, 560, and 385 nm and blue, green, and UV filter sets for sample analysis in fluorescence mode; (iii) color digital Camera Optika C-P6, 6.3 MP; and (iv) OPTIKA PRO VIEW (Optika Srl, Ponteranica, Italy) software for image acquisition and processing. Samples were characterized with 10× magnification objectives in transmitted illumination mode.
4
Microscopy Characterization of Microspheres and HPM Composites
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Microspheres and HPM composites were characterized with an IM-5FLD inverted fluorescence microscope (Optika Srl, Ponteranica, Italy) equipped with (i) an 8W XLED illumination source for sample analysis under transmitted light; (ii) 5W LED excitation illumination sources at 470, 560, and 385 nm and blue, green, and UV filter sets for sample analysis in fluorescence mode; (iii) color digital Camera Optika C-P6, 6.3 MP; and (iv) OPTIKA PRO VIEW (Optika Srl, Ponteranica, Italy) software for image acquisition and processing. Samples were characterized with 10× magnification objectives in transmitted illumination mode.
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