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Live-Cell Analysis Handbook — Third Edition

Kinetic Neuroimmune Assays

Real-time quantification of chemotaxis and
phagocytosis to evaluate neuroimmune function

Introduction

Several neurological disorders have been linked with the morphology of activated cells, their migratory response, and the
dysfunction of the immune system, including multiple sclerosis, ability to phagocytize diseased or dying cells as well as pathogens,
Alzheimer’s disease and brain cancers. Being the primary innate employing technologies such as flow cytometry and Boyden
immune cells of the brain, microglia, play a role in both the chamber assays. However, the techniques used to study microglia
etiology of such disease progression as well as in heathy brain behavior is typically endpoint, are performed in environments
development and maintenance. In their role as resident brain that are not representative of physiological conditions, include
macrophages, microglia are responsible for immunosurveillance cumbersome assay preparation steps and do not offer insight into
and neuroprotection, regulating brain development primarily morphological changes associated with microglial activation.
through phagocytosis and the release of various immune proteins.
However, with age, microglia become increasingly dysfunctional Live-cell imaging and analysis addresses these inherent drawbacks
and lose their neuroprotective properties. Thus, studying the via non-invasive, repeated monitoring of the same population
role of these innate immune cells is crucial in understanding the of cells within a standard cell culture incubator. The includes
inflammatory response under both normal and degenerative incorporation of reagents and consumables that allow for real-
conditions of the brain. time, quantification and visualization of microglial behavior,
alleviating technically challenging preparation and quantification
To evaluate microglia behavior and their impact on the regulation steps associated with traditional techniques. In this chapter, we
of functional neurons, detailed in vitro methodology is required. will examine kinetic approaches that can shed new light on the
In particular, the cell models used, such as microglia cell lines, function of microglia in order to characterize microglial response
stem cell-derived microglia cultures and primary dissociated to neurodegeneration therapeutic interventions.
cell cultures, as well as their culture conditions is of pivotal
importance to characterize the underlining functions of microglia.
Assays designed to evaluate these models typically focus on the

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