Application

Combined Live Cell and Flow Cytometry Analysis of Immune Cell Killing

IncuCyte-iQue Workflow

In vitro cell biology models are becoming ever more complicated in order to better mimic in vivo studies. However deciphering the meaning of results gained from the complex biological interactions becomes more difficult and requires a greater understanding of the biology. Combining specialist technologies provides the user with more detailed analysis in order to look deeper into the biology and make more physiologically relevant conclusions. In order to do this, workflows must be developed that link technology platforms using a streamlined approach that is simple to follow as well as using both time and samples efficiently.
 

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IncuCyte-iQue Workflow Concept

  • The IncuCyte-iQue workflow uses the immune cell killing application to describe how a single assay plate can be used to acquire both real time data, daily cytokine sampling and T cell activation panel
  • Adherent or non-adherent target cells expressing NucLight Green are co-cultured with immune cells (label free)
  • Tumour cell proliferation and death are monitored using the fluorescent green signal from the transduced cells and the addition of Annexin V to the culture
  • Optional: Cytokine analysis is monitored throughout experiment by sampling from assay plate and assaying using Qbeads kit
  • Automated image analysis enables selective quantitation of tumor cell death (basic analyser) as well as PBMC proliferation and shape change (cell-by-cell).
  • Cells are washed lifted (following the IncuCyte-iQue Workflow protocol) and labelled using the TCA kit
  • T cell subset and activation are then quantified using a pre-defined gating strategy

Key Advantages

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Combined subset & cytokine analysis with spatial and morphological quantification

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Easy to use, mix-and-read assays using Intellicyt-IncuCyte compatible reagents

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Kinetic cytokine measurements using non-perturbing low volume supernatant sampling maximising sample usage with minimal manipulation

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Additional biological insight providing a greater depth of physiological understanding

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Combined subset analysis with spatial and morphological quantification

 

Easy to use, mix-and-read assays using Intellicyt-IncuCyte compatible reagents

 

Kinetic cytokine measurements using non-perturbing low volume supernatant sampling maximising sample usage with minimal manipulation

 

Additional biological insight providing a greater depth of physiological understanding

 

Ordering Information

QBeads Assay Builder
IFNg option 1qbeads assay builder         
TNFα option
T Cell Activation Cell and Cytokine Profiling Kit 
PlatformCompatible with iQue3 VBR configuration
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Available SizesCatalog Numbers
1 x 96 well
90560
t cell activation cell and cytokine profiling kit
5 x 96 wells
90561
1 x 384 wells
90562
5X384 wells
90563