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. 2016 May 16;6:26013. doi: 10.1038/srep26013

Figure 1. Demonstration of passive CLARITY and optimisation of immunofluorescent labelling conditions on wild type mouse brain sections.

Figure 1

Representative images of wild type 12 month old C57/Bl6 mouse cerebellar sections of varying thickness are shown pre- and post-passive clearance (a). Passively cleared cerebellar sections were immunofluorescently labelled with antibodies raised against porin (green), neurofilament H (NF-H; red) and myelin basic protein (MBP; purple). Various conditions were tested for the immunolabelling protocol; (b) sodium borate buffer at 37 °C for 24 hours, (c) sodium borate buffer at 4 °C for 6 days for the primary antibodies, then at 4 °C for 4 days for secondary antibodies, (d) PBST at 4 °C for 6 days for the primary antibodies, then at 4 °C for 4 days for secondary antibodies and (e) The advantages of passively clearing tissue sections is exemplified in an uncleared section which was stained using the protocol in (d). Scale: 100 μm.