NETosis

Neutrophils play a key role in the body’s defense against infections. As part of the innate immune system, they employ multiple strategies to degrade and kill microbes, including the release of neutrophil extracellular traps (NETs). NETs are a web-like structure composed of antimicrobial proteins and DNA that are released during a distinct form of programmed neutrophil cell death known as NETosis. In general, NETs allow neutrophils to kill extracellular pathogens while minimizing damage to the host cells but can also result in an autoimmune response. This process in vivo is not well understood, so in vitro models are important to help clarify the mechanisms and results of this phenomenon.

 

Netosis Assay Procedure

Assay Setup

Step 1. Assay setup: Neutrophils preloaded with plasma membrane integrity stain provide an assay to kinetically monitor NETosis by imaging in microplates

 

Assay Setup

Step 2. Automated kinetic imaging: Kinetic fluorescence imaging of stimulated cells captures release of NETs over time.

 

Assay Setup

Step 3. Automated analysis: Treatment-dependent NETosis is quantified using object masking and fluorescence integral.

Increase assay throughput and performance

Agilent BioTek BioSpa compatible assay performs live cell analysis on up to 8 microplates. The Agilent BioTek BioSpa live cell imaging system enables multiplate kinetic analysis of the NETosis assay.
BioSpa
Figure 1. The Agilent BioTek BioSpa live cell analysis system

Liquid handling automation with the Agilent BioTek MultiFlo FX multimode dispenser or other liquid handlers can automate cell seeding and treatment addition.

BioSpa

Figure 2. The Agilent BioTek MultiFlo FX multimode dispenser

 

Resources

 

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RA44218.4343981482

 

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