Cell-Mediated Cytotoxicity

Immune-cell-mediated cytotoxicity plays an essential role in defending the host from intracellular pathogens. Activated effector cells, including T cells, NK, natural killer T cells (NKT), and granulocytes, are capable of killing targeted cells through unique mechanisms and regulatory pathways. Kinetic imaging and analysis of fluorescently labeled target and effector cells provides an effective and convenient method for characterizing cell-mediated cytotoxicity over time. This method can be used in both 2D and 3D cell culture formats.

 

Cell-Mediated Cytotoxicity Assay Procedure

Assay Setup

Step 1. T Cell Activation: Labeled T cells (red) attack target cells in media containing fluorescent cytotoxicity probe.

 

Automated kinetic fluorescence imaging

Step 2. T Cell-Induced Cytotoxicity: Kinetic brightfield and fluorescence imaging of cytotoxicity probe (green) reveals T cell-induced cytotoxicity in target cell

 

Automated-analysis

Step 3. Automated Analysis: Detailed profiles of necrosis probe fluorescence over time demonstrate T cell-mediated cytotoxicity.

Increase assay throughput and performance

Agilent BioTek BioSpa automated incubator performs live cell compatible assay with live cell analysis on up to 8 microplates. The BioSpa live cell analysis system enables multi-plate kinetic analysis of the T cell activation and proliferation assay.
BioSpa
Figure 1. The BioSpa live cell analysis system

Liquid handling automation with the Agilent BioTek MultiFlo FX multimode dispenser or other liquid handler can automate seeding cells into the microplate and conduct media exchanges.

 

BioSpa

Figure 2. The MultiFlo FX multimode dispenser

 

Resources

 

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