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Kinetic Neuroimmune Assays
Automated quantification of microglia migration
In addition to providing insight into the morphology of microglia, fragment involved in cell recruitment and activation of
the IncuCyte can also be utilized to quantify migratory capability phagocytosis. After cell addition on the top membrane of the
of these cells via IncuCyte Chemotaxis Assays. Microglia are the IncuCyte ClearView chemotaxis plate, three-fold dilutions of C5a
resident immune cells of the brain, responding to extracellular were added to reservoir wells. Cells then migrated to the bottom
signals and migrating towards the site of infection or neuronal of the membrane toward C5a. Label-free measurements of
damage. In a model of microglia migration response, iPSC- bottom-side confluence were analyzed in real time. Data shows
derived microglia (CDI) were seeded in an IncuCyte ClearView concentration-dependent chemotactic migration of iPSC-derived
96-well Chemotaxis Plate and exposed to C5a, a protein microglia towards C5a over a period of 24 hours (Figure 2).
Top of membrane with mask Bottom of membrane with mask
Concentration-dependent movement towards
complement component 5a
Phase area bottom
2
(μm /well)
4 x 10 5
1000 nM
3 x 10 5
2 x 10 5 333 nM
1 x 10 5 111 nM
37 nM
12 nM
4 nM
0 Control
0 8 16 24
Hours
Figure 2. Concentration-dependent migration of iPSC-derived microglia (CDI) toward C5a. IPSC-derived microglia (CDI) were plated in the top chamber of
the IncuCyte ClearView 96-Well Cell Migration Plate at a density of 4,000 cells/well. Serial dilutions of C5a, starting at 1 nM, were added to the bottom
®
reservoir wells and automated imaging and analysis was performed (data collected at 1-hour intervals). Images represent the top and bottom side of the
membrane at the 30-hour time point. Automated image processing separates cells located on the top (outlined in yellow) and the bottom (outlined in
black) surface of the membrane. Pores are outlined in white. Images are processed as they are acquired, and data can be plotted in real time.
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