CD-1^® mouse brain tissue postnatal day 5 was dissociated using the Neural Tissue Dissociation Kit (T) and the gentleMACS™ Dissociator. Brain cells were intracellularly stained with Anti-GFAP antibodies or with the corresponding REA Control (I) antibodies (left images) as well as with Anti-GLAST (ACSA-1) antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

3D-immunofluorescence analysis of a mouse brain hemisphere fixed with 4% PFA and subjected to whole-mount immunostaining with GFAP Antibody, anti-human/mouse/rat, Vio R667 (red). The sample was processed and optically cleared using the MACS^® Clearing Kit and imaged with an UltraMicroscope II light sheet instrument. Sample detail image of cortex at approximately 100µm depth with sagittal section and lateral-medial orientation (A) and overview image (B). Scale bars: 100 µm (A), 500 µm (B).

3D-immunofluorescence analysis of an adult mouse brain hemisphere fixed with 4% PFA and subjected to whole-mount immunostaining with GFAP Antibody, anti-human/mouse/rat, Vio R667 (red). The sample was processed and optically cleared using iDISCO+ clearing (for protocol details please see herehttps://idiscodotinfo.files.wordpress.com/2015/04/whole-mount-staining-bench-protocol-methanol-dec-2016.pdf and imaged with an UltraMicroscope Blaze light sheet instrument. Imaging data was digitally post-processed. Sample detail image (A), overview image (B), 3D-rendered maximum intensity projection (C) and 3D-rendered video (D). Scale bars: 100 µm (A), 500 µm (B), 500 µm (C).