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
Step 1. T Cell Activation: Labeled T cells (red) attack target cells in media containing fluorescent cytotoxicity probe.
Step 2. T Cell-Induced Cytotoxicity: Kinetic brightfield and fluorescence imaging of cytotoxicity probe (green) reveals T cell-induced cytotoxicity in target cell
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.
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.
Figure 2. The MultiFlo FX multimode dispenser
- An Image-Based Method to Detect and Quantify T Cell Mediated Cytotoxicity of 2D and 3D Target Cell Models
For Research Use Only. Not for use in diagnostic procedures.