Figure 4
Histone-GFP labeled cell nuclei. (a–c) Confocal Microscopy of fixed brain slices of Histone-GFP mice shows that all DAPI-stained nuclei (a) are also labeled with Histone-GFP (b) as seen in the overlap (c, green = Histone-GFP, magenta = DAPI). Scale bar = 10 µm. (d) Representative example image consisting of a maximum intensity projection of 10 consecutive image frames covering 20 µm of tissue recorded at a depth of 200 µm in Histone-GFP positive mice using 2Pii. Scale bar = 100 µm. (e) Close up of D. Scale bar = 20 µm. (f) Average intensity projection of a stack cropped in ZX. The surface of the brain is located at the top of the image. Red stars delineate the lower border of cortical layer 1 as visible in change of cell density. Scale bar = 50 µm. (g) A representative rendering of a 700 µm × 700 µm × 700 µm stack recorded by in vivo 2Pii in a Histone-GFP mouse. One orthogonal slice is displayed in the middle of the stack. The red stippled line denotes the ‘shadow’ cast by a blood vessel which strongly attenuates the laser beam. (h) Detected nuclei within a volume of 700 µm × 700 µm × 700 µm. Each sphere corresponds to the centroid position of a nucleus and is randomly coloured.
