Microglia Phagocytosis Assay

Microglia are the resident immune-effector cells in the central nervous system and play a critical role in the immune response of the brain. Microglial activation typically occurs in response to neurological damage such as brain injury, neurodegeneration, and infection, which is thought to play a key role in the progression of neurodegenerative diseases.

Phagocytosis, in response to neuronal damage and infectious agents, is a key function of microglia for the clearance of apoptotic cells, extracellular protein aggregates, and infectious bacteria in the central nervous system (CNS). Neuronal homeostasis and synaptic pruning are supported by microglia through phagocytosis. Dysfunctional macrophage phagocytosis has been implicated in many inflammatory conditions and can lead to aberrant disease materials such as dysfunctional cells or aggregated proteins such as α-Synuclein and β-amyloid.

Fig.1 Immunocytochemistry illustrates phagocytosis of fluorescent latex beads (yellow-green) by microglia (red, labelled with anti-OX42). (Pul, et al., 2013)

Microglia Phagocytosis Assay at Creative Biolabs

We use a series of pHrodo-labeled baits, which are pH-sensitive dye conjugates that fluoresce brightly in phagosomes and can label proteins, beta-amyloid (1-42), alpha-synuclein, and dead cells, making the assay highly customizable. The fluorescence of pHrodo BioParticles increases as pH decreases from neutral to acidic. No washing or quenching steps, or compensation for environmental factors, ensures faster, more accurate results.

Our IncuCyte S3 Live Cell Analysis System is a simple and flexible analysis platform that provides long-term kinetic measurements and real-time cell visualization. The ability to automatically acquire and analyze live cell images provides continuous analysis of cells in situ in physiologically relevant settings.

Microglia Phagocytosis Assays Methodology

1. Microglia are isolated from adult mice and purified in defined culture conditions.
2. Preparing pHrodo Red S. aureus BioParticles Conjugate.
3. Pre-incubated with the therapeutic agents.
4. Phagocytosis is kinetically monitored using the IncuCyte S3 platform.

Sample Data

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Reference

1. Pul, R.; et al. Quantification of microglial phagocytosis by a flow cytometer-based assay. Microglia. Humana Press, Totowa, NJ, 2013: 121-127.