Cv1000 confocal microscope

For all experiments where live cell imaging was used, OECs were plated onto glass culture plates (6×10³ cells/chamber in 8-well glass chamber slides) and imaged in CO₂ independent medium (Invitrogen) in a chamber maintaining cells at 37°C. Epifluorescent live cell imaging was carried out using a Hamamatsu digital camera on an Olympus IX81 CellR microscope fitted with epifluorescence and differential contrast optics with time-lapse images taken every 10 min for up to 24 h. For some assays, confocal live cell imaging was carried out on an Olympus CV1000 confocal microscope with images taken every 3 min.

To obtain cortical slices covered with intact meninges, whole forebrains isolated from E14 male and female CX3CR1-GFP mice that had been electroporated at E12 were embedded in 2% agarose gel, and then sliced coronally (350 µm) using a vibratome. The slices were cultured in collagen gel as previously described (Miyata et al., 2004 (link)). Time-lapse imaging was performed on a CV1000 confocal microscope (Olympus). Chambers for on-stage culture were filled with 40% O₂.