Incucyte basic software v2018a

Manufactured by Sartorius

The IncuCyte™ Basic Software v2018A is a software tool that provides core functionality for the IncuCyte live-cell analysis system. It enables users to capture, analyze, and manage data from live-cell imaging experiments.

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Lab products found in correlation

Caspase 3 7 green dye, by Sartorius (2 mentions) Celltrace far red, by Thermo Fisher Scientific (1 mentions) Poly l ornithine, by Merck Group (1 mentions) Tissue culture treated microplates, by Corning (1 mentions) Ula plate, by Corning (1 mentions)

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IncuCyte (373 citations) IncuCyte software (87 citations) IncuCyte Basic Software (2 citations)

3 protocols using incucyte basic software v2018a

Real-time Cytotoxicity Assay for B16 Melanoma

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Killing of B16 melanoma cells was evaluated in real-time using the IncuCyte platform. Activated WT or P2rx7^−/− P14 CD8⁺ T cells were plated into 96-well flat clear-bottom tissue culture–treated microplates, previously coated with Poly-L-ornithine (Sigma Aldrich #A-004-M) for 2–3 hours, along with CellTrace Far Red (Thermo Fisher Scientific #C34564) stained (according to manufacturer’s instructions) B16.gp33 or B16.OVA cells at 1:1 and 3:1 effector:target ratios. 5uM of Caspase-3/7 green dye (Sartorius #4440) was added just prior to beginning assay to identify dying cells. The plate was then placed in an IncuCyte ZOOM platform housed inside a cell incubator at 37°C/5% CO₂. Images from 5 technical replicates were taken every 30 minutes for 48 hours using a 4X objective lens and then analyzed using IncuCyte Basic Software v2018A (Sartorius). Graphed readouts represent percentage of live B16 melanoma targets (CellTrace Far Red⁺Caspase-3/7⁻), normalized to live targets alone at the starting (0 hour) time point.

Wanhainen K.M., Peng C., Zhou M.H., de Gois Macedo B., O’Flanagan S., Yang T., Kelekar A., Burbach B.J., da Silva H.B, & Jameson S.C. (2022). P2RX7 enhances tumor control by CD8+ T cells in adoptive cell therapy. Cancer immunology research, 10(7), 871-884.

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Real-time Tumor Cell Killing Assay

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Tumor killing was evaluated in real-time using the IncuCyte platform. Magnetic-bead-enriched CD3-CD56+ NK effector cells were plated into 96-well flat clear-bottom polystyrene tissue-culture-treated microplates (Corning, Flintshire, UK) along with NuclightRed stably expressing OVCAR8 cells at a 2:1 effector:target ratio. Caspase-3/7 green dye (Sartorious, Ann Arbor, MI, USA) was added to pick up dying cells that have not yet lost NuclightRed fluorescence. Noted treatments were then added at a 30 nM concentration, and the plate was placed in an IncuCyte ZOOM^® platform housed inside a cell incubator at 37 °C/5% CO₂. Images from three technical replicates were taken every 15 min for 48 h using a 4X objective lens and then analyzed using IncuCyte™ Basic Software v2018A (Sartorious). Graphed readouts represent percentage live OVCAR8 targets (NuclightRed⁺Caspase-3/7⁻), normalized to live targets alone at the starting (0 h) time point.

Vallera D.A., Ferrone S., Kodal B., Hinderlie P., Bendzick L., Ettestad B., Hallstrom C., Zorko N.A., Rao A., Fujioka N., Ryan C.J., Geller M.A., Miller J.S, & Felices M. (2020). NK-Cell-Mediated Targeting of Various Solid Tumors Using a B7-H3 Tri-Specific Killer Engager In Vitro and In Vivo. Cancers, 12(9), 2659.

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Real-Time Tumor Spheroid Killing Assay

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Tumor killing was evaluated in real-time using the IncuCyte S3 platform. A total of 20,000 SKOV3-GFP cells/well were plated in 96-well round-bottom ultra-low adhesion (ULA) plates (Corning, Flintshire, UK) and allowed to form spheroids for 3 days. A total of 40,000 magnetic-bead-enriched NK cells were then added to each well. Noted treatments were then added at a 30 nM concentration, and the plate was placed in an IncuCyte S3^® platform housed inside a cell incubator at 37 °C/5% CO₂. Images from three technical replicates were taken every 45 min for 72 h using a 4× objective lens and then analyzed using IncuCyte™ Basic Software v2018A (Sartorious). Graphed readouts represented target spheroid size and intensity, normalized to live spheroids alone at the starting (0 h) time point.

Vallera D.A., Oh F., Kodal B., Hinderlie P., Geller M.A., Miller J.S, & Felices M. (2021). A HER2 Tri-Specific NK Cell Engager Mediates Efficient Targeting of Human Ovarian Cancer. Cancers, 13(16), 3994.

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