The mice were anesthetized with chloral hydrate (10% W/V, 500 mg/kg body weight, i.p.), and perfused transcardially with PBS (5 min), followed by ice-cold 4% paraformaldehyde (PFA, 158127 MSDS, sigma) dissolved in PBS (5 min). The brain tissues were carefully removed and post-fixed in PBS containing 4% PFA at 4°C overnight, and then equilibrated in PBS containing 25% sucrose at 4°C for 72 h. The 40 μm thick coronal slices of the whole brain were obtained using the cryostat microtome and stored at -20°C.
For RV or AAV labeled samples, every sixth section of the brain slices were selected, stained with DAPI, washed with PBS, mounted with 90% glycerol (in PBS) and sealed with nail polish.
For PRV-152 labeled samples, the procedures for immunohistochemistry were performed as before (Wei et al., 2015 (link)). Every sixth section of the brain slices were selected and stained with GFP (abcam, ab290, 1: 1000) and DAPI, and then mounted and sealed as described above.
All of the images were captured with the TCS SP8 fluorescence laser scanning confocal microscope (Leica, China) or the Olympus VS120 virtual microscopy slide scanning system (Olympus, Shanghai, China).
For RV or AAV labeled samples, every sixth section of the brain slices were selected, stained with DAPI, washed with PBS, mounted with 90% glycerol (in PBS) and sealed with nail polish.
For PRV-152 labeled samples, the procedures for immunohistochemistry were performed as before (Wei et al., 2015 (link)). Every sixth section of the brain slices were selected and stained with GFP (abcam, ab290, 1: 1000) and DAPI, and then mounted and sealed as described above.
All of the images were captured with the TCS SP8 fluorescence laser scanning confocal microscope (Leica, China) or the Olympus VS120 virtual microscopy slide scanning system (Olympus, Shanghai, China).
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