1. Microglia identity was verified using RT-qPCR using a panel of microglia-specific and macrophase-specific markers. RT-qPCR results showed that several microglial mRNA markers were upregulated in the matured microglia, while stemcell markers were absent.
2. To test the phagocytic function of the iPSC-derived microglia, the cells were tested for their ability to phagocytose exogenous materials. Microglia were able to phagocytose bacterial like bioparticles and Amyloid oligomers/fibrils (see figure 6 showing the uptake of fluorescently labeled material over time). Bioparticle uptake correlated well with PRS (figure 7) however, this was mainly driven by the highly phagocytosing APOE4/4 carrying microglia.
3. reMYND was also able to monitor morphology and motility using live brightfield imaging of microglia. In these assays microglia are individually tracked over time after LPS induced activation. Using machine learning models, phenotypes were characterized and analyzed. 

For long-term impedance monitoring, the measurement system was placed in the incubator to keep the microglia in their optimal environment during recordings (Figure 9). For combined microscope imaging and impedance monitoring, the measuring system was placed on the microscope stage. Moreover, a controlled environment of 37°C and 5% CO2 was created around the microscope.