Figure 1.

Three-dimensional fluorescence imaging of microglial activation in response to neurodegeneration. (A–E) 3D fluorescence imaging of the mouse optic nerves. (A) The optic nerves before (upper panel) and after (lower panel) the iDISCO procedure. (B–D) The optic nerves of wildtype mice were processed for the whole-tissue immunolabeling of PECAM1 (B), LYVE1 (C) or CD11b (D) and imaged on the lightsheet microscope. (E) The optic nerves of Aldh1l1-GFP transgenic mice were processed for the whole-tissue immunolabeling of GFP and imaged on the lightsheet microscope. Representative orthogonal (upper panels) or perspective (lower panels) 3D-projection images of the optic nerves are shown. (F) 3D fluorescence imaging of the traumatic injury-induced neurodegeneration. The wildtype mice were intravitreally injected with the tdTomato-expressing AAV2 and then subjected to optic nerve injury. The control (i.e., uninjured) and injured nerves were processed for the whole-tissue immunolabeling of tdTomato and imaged on the lightsheet microscope. Representative orthogonal (upper panels) or perspective (lower panels) 3D-projection images of the optic nerves are shown. (G and H) 3D fluorescence imaging of the glial responses to neurodegeneration. The wildtype mice were subjected to optic nerve injury. The control and injured nerves were processed for the whole-tissue immunolabeling of Iba1 (G) or Olig2 (H) and imaged on the lightsheet microscope. Representative orthogonal (upper panels) or perspective (lower panels) 3D-projection images of the optic nerves are shown