The possible nuclear damage and apoptotic changes caused by exposure of colon cancer cells to Fe 3 O 4 @Glu-Oleuropein NPs were studied by the Hoechst staining. In brief, the SW480 cells were treated with the nanoparticles for 24 h, and next, were stained with the Hoechst 33258 solution. Finally, the cells were washed with PBS and examined under a fluorescent microscope (Incell Analyser 2000, USA).
Analyser 2000
Manufactured by INCELL
Sourced in United States
The Incell Analyser 2000 is a laboratory instrument designed for the analysis of cellular samples. It is capable of performing a variety of measurements and assessments on cell cultures, tissues, or other biological samples. The core function of the Incell Analyser 2000 is to provide quantitative data on cellular parameters such as cell count, viability, proliferation, and morphology.
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Lab products found in correlation
6 protocols using analyser 2000
1
Evaluating Nanoparticle-Induced Apoptosis in Colon Cancer
2
Clonogenic Assay for Colony Quantification
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Clonogenic assay is a versatile tool for in vitro screening of the capacity of a single-cell suspension to form a colony of 50 or more cells under different circumstances. The assay was made according to Franken et al. 26 . A total of 100 cells were seeded into 6-well plates containing standard culture media. After 10 days, colonies were stained and the well images captured in the InCell Analyser 2000 Automated uorescence wide eld HCS microscope. Colony number and respective areas were measured using the colony count plugin from ImageJ software 59 .
3
Hoechst Nuclear Staining of Colon Cancer Cells
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To accomplish the Hoechst nuclear staining, colon cancer cells were treated with Ag@Gln-TSC NPs for 24 h (at IC50 concentration) and then, stained with the Hoechst 33,258 solution (Sigma-Aldrich, USA). Subsequent, the cells were washed with PBS, examined under a fluorescent microscope, and compared with control cells (Incell Analyser 2000, USA).
4
Cell Migration Wound Healing Assay
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Cells were seeded in 24-well plates at a density of 2 × 105 cells/well in 500 µL of Dulbecco’s modified Eagle’s medium, DMEM 1X (Gibco, Paisley, UK) supplemented with 5% (v/v) Charcoal Stripped Fetal Bovine Serum Qualified One Shot™ (Gibco, Monterrey, Mexico), for cells mitosis synchronization, and 1× antibiotic solution penicillin-streptomycin (pen-strep, Grand Island, NE, USA). After wound, cells were incubated for 15 h, and images were captured every 3 h in the InCell Analyser 2000 Automated fluorescence widefield HCS microscope. The extent of wound closure was measured using the MRI wound-healing tool from ImageJ software.
5
Evaluating Nuclear Damage in AGS Cells
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The nuclear damages caused by CuO@Glu/TSC NPs in treated AGS cells were evaluated by the Hoechst staining method [11] (link). At rst, the AGS cells (4×10 5 cells/well) were prepared and treated with CuO@Glu/TSC NPs at IC 50 concentration. The AGS cells without NPs exposure were regarded as control cells. After 24 h incubation, the cells were stained with Hoechst 33258 dye and visualized using a uorescent microscope (Incell Analyser 2000, USA).
6
Wound Healing Assay Automation
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Cells were seeded in 24-well plates at a density of 2 × 10 5 cells/well in 500 µl of Dulbecco's modi ed Eagle's medium, DMEM 1X (GIBCO, Paisley, UK) supplemented with 5% (V/V) Charcoal Stripped Fetal Bovine Serum Quali ed One Shot™ (Gibco, Mexico), for cells mitosis synchronization, and 1X antibiotic solution penicillin-streptomycin (pen-strep, Gibco™). At least 48 h post-seed, to ensure cell synchronization and growth to 90%-100% con uence, a single scratch wound was made in each well using a 200 µl disposable pipette tip. The cells were then incubated for 15 h in the InCell Analyser 2000 Automated uorescence wide eld HCS microscope for the capture of wound images every 3 h. The extent of wound closure was measured using the MRI wound healing tool from ImageJ software 59 .
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