Figure 18B. Analysis of time-lapse confocal images of microglia in 2D and 3D culture

Representative confocal time-lapse images of microglia culture in 3D over 10 minutes.
We investigated microglial motility in 3D culture. To achieve this, we obtained time-lapse images of microglia using a high-resolution spinning disk confocal microscope equipped with a 60x water immersion objective. A time frame of 30s per image over 10 min with a z-slice interval of 0.5 µm was used to record the motility of microglia in 2D and 3D cultures. We observed that microglia in 3D culture showed a ramified morphology with constant extensions and retractions of processes reminiscent of in vivo-like microglial surveillance (Figure 18A and B). In contrast, microglia in 2D culture showed an amoeboid morphology with membrane ruffling (Figure 18B ).