Fig. 2

Immobilizing non-adherent GL261 cell neurospheres in low melting point agarose (LMPA) for live-cell fluorescent (calcium) imaging. a A 20 μL droplet containing suspended cells was applied to the bottom of the imaging 8-well chamber slide. b 20 μL of LMPA was placed on top of this droplet. LMPA is liquid at 37 °C (culture temperature of the cells) but solidifies upon cooling to 26–30 °C (room temperature at which imaging is performed). c Extracellular solution is placed on top of cells and LMPA. Test compounds can be delivered to the extracellular solution without disturbing cells, and solutes diffuse freely through the LMPA