In vitro differentiated murine MDSC (1 x 106 cells) stained with Hoechst (20 μM, ThemorFisher) and calcein (1mg/ml diluted 1:2000, Invitrogen) were co-cultured with the indicated tumor cells (ANV5, 4T1 or derivatives), previously stained with orange-CMRA (1mg/mL, diluted 1:2000, Invitrogen) and Hoechst. The coculture was embedded at 103 cells/μL in hydrogels. After 48 h (for mouse cells) and 20 h (for human cells), images were captured by a confocal microscope LSM 800 laser-scanning (Carl Zeiss) with Plan-apochromat-63x. Images in Z were acquired with Zen 2.3 software (Carl Zeiss) and were visualized with Volocity 3D (RRID:SCR_002668, ThermoFisher Scientific). NET area and the percentage of NETs was calculated using the plugin DANA for ImageJ (developed by Dr. Miriam Shelef). At least 60 cells per experimental condition were analyzed (59 (link)). Experiments were repeated three times.
Volocity 3d
Manufactured by Thermo Fisher Scientific
Volocity 3D is a software solution designed for high-resolution 3D imaging and analysis. It provides advanced tools for visualizing, analyzing, and managing large volumes of image data from a variety of microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst, by Thermo Fisher Scientific (3 mentions)
Lsm 800 laser scanning, by Zeiss (2 mentions)
Zen 2, by Zeiss (2 mentions)
Orange cmra, by Thermo Fisher Scientific (1 mentions)
Plan apochromat 63x, by Zeiss (1 mentions)
Calcein, by Thermo Fisher Scientific (1 mentions)
Plan apochromat 63, by Zeiss (1 mentions)
2 protocols using volocity 3d
1
Quantifying Neutrophil Extracellular Traps
2
Murine MDSC-Tumor Cell Coculture Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro–differentiated murine MDSC (1 × 106 cells) stained with Hoechst (20 µmol/L, Thermo Fisher) and calcein (1 mg/mL diluted 1:2,000, Invitrogen) were cocultured with the indicated tumor cells (ANV5, 4T1, or derivatives), previously stained with orange CMRA (1 mg/mL, diluted 1:2,000, Invitrogen) and Hoechst. The coculture was embedded at 103 cells/µL in hydrogels. After 48 hours (for mouse cells) and 20 hours (for human cells), images were captured by a confocal microscope LSM 800 laser-scanning (Carl Zeiss) with Plan-apochromat-63×. Images in Z were acquired with Zen 2.3 software (Carl Zeiss) and were visualized with Volocity 3D (RRID:SCR_002668, Thermo Fisher Scientific). NET area and the percentage of NETs were calculated using the plugin DANA for ImageJ (developed by Dr. Miriam Shelef, University of Wisconsin–Madison, Madison, WI). At least 60 cells per experimental condition were analyzed (59 (link)). Experiments were repeated three times.
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In vitro–differentiated murine MDSC (1 × 106 cells) stained with Hoechst (20 µmol/L, Thermo Fisher) and calcein (1 mg/mL diluted 1:2,000, Invitrogen) were cocultured with the indicated tumor cells (ANV5, 4T1, or derivatives), previously stained with orange CMRA (1 mg/mL, diluted 1:2,000, Invitrogen) and Hoechst. The coculture was embedded at 103 cells/µL in hydrogels. After 48 hours (for mouse cells) and 20 hours (for human cells), images were captured by a confocal microscope LSM 800 laser-scanning (Carl Zeiss) with Plan-apochromat-63×. Images in Z were acquired with Zen 2.3 software (Carl Zeiss) and were visualized with Volocity 3D (RRID:SCR_002668, Thermo Fisher Scientific). NET area and the percentage of NETs were calculated using the plugin DANA for ImageJ (developed by Dr. Miriam Shelef, University of Wisconsin–Madison, Madison, WI). At least 60 cells per experimental condition were analyzed (59 (link)). Experiments were repeated three times.
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