IncuCyte® Immune Cell Killing FAQs

Immuno-Oncology

Yes, as long as the fluorophore excitation and emission spectra are compatible with the IncuCyte ZOOM® fluorescence imaging channels (green channel: excitation 440–480 nm, emission 504–544 nm; red channel: excitation 565–605 nm, emission 625–705 nm). Note that data quality may be compromised if fluorophore expression is unstable over time or results in heterogeneous labeling. We recommend our NucLight™ Red labeling reagent (Essen BioScience 4476), as it is ideally suited for long-term livecell studies. The NucLight™ Red labeling reagent does not disturb cell health and is expressed stably and homogeneously

Yes. Please see our protocol for suspension target cell lines.

No. Cytoplasmic cell labels (e.g., CytoLight™ Red; Essen BioScience 4482) can be used to label target cells; however, you will not be able to resolve and count individual target cells. Instead, cell area metrics such as Red Object Area or Red Confluence would be used to measure proliferation.

There is no need to pre-label your target cells when using adherent tumor cells. Tumor cell death is detected using the IncuCyte™ Caspase-3/7 Apoptosis Reagent (Essen BioScience 4440), which is simply added to your assay along with your treatments. Although there is no need to label your target cells, you will find that using our NucLight™ Red labeling reagent (Essen BioScience 4476) to label your tumor cells will enable you to make simultaneous measurements of viable target cell count in real time. If you are using suspension target cells, they will need to be prelabeled. Measurements of non-adherent tumor cell killing are made by quantifying the reduction in target-cell fluorescence over time.

Yes. Target-cell proliferation can still be measured if your tumor cells are labeled with a green fluorophore. However, if a green label is used for your target cells, you will need to use a red fluorescent probe to measure target cell death. Please refer to the IncuCyte ZOOM® Fluorescent Dye Optimization Technical Note for options around suitable red cell death probes.

Yes. However, the probe must be suitable for use in long-term live cell assays. Please refer to the IncuCyte ZOOM® Fluorescent Dye Optimization Technical Note for options around suitable probes.

Yes. We have successfully performed immune cell killing experiments in a 384-well format.

Yes. Simply harvest cells/supernatants when the assay is complete, and perform your choice of further analysis.

Yes. However, because the Incucyte™ FLR has only has one imaging channel, you will only be able to make measurements using the green fluorescence channel.

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