Histology was conducted in the same manner as previously reported (Tsutsui-Kimura et al., 2020 (link)). Mice were perfused using 4% paraformaldehyde, then brains were sliced into 100μm thick coronal sections using a vibratome (Leica) and stored in PBS. These slices were then stained with rabbit anti-tyrosine hydroxylase (TH; AB152, EMD Millipore, RRID: AB_390204) at 4°C for 2d to reveal dopamine axons in the striatum, dopamine cell bodies in the midbrain, and other neurons expressing TH throughout the brain. Slices were then stained with fluorescent secondary antibodies (Alexa Fluor 594 goat anti-rabbit secondary antibody, A-11012, Invitrogen, RRID: AB_2534079) at 4°C for 1d. Slices were then mounted in anti-fade solution (VECTASHIELD anti-fade mounting medium, H-1200, Vector Laboratories, CA) and imaged using Zeiss Axio Scan Z1 slide scanner fluorescence microscope (Zeiss, Germany).
Axio scan z1 slide scanner fluorescence microscope
Manufactured by Zeiss
Sourced in Germany
The Zeiss Axio Scan Z1 is a slide scanner and fluorescence microscope. It is capable of digital scanning and imaging of microscope slides.
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2 protocols using axio scan z1 slide scanner fluorescence microscope
1
Dopaminergic Neuron Immunohistochemistry
2
Immunohistochemical Analysis of Dopaminergic Neurons
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Histology was conducted in the same manner as previously reported (Akiti et al., 2021; Menegas et al., 2018) . Mice were perfused using 4% paraformaldehyde, then brains were sliced into 100 µm thick coronal sections using a vibratome (Leica, Germany) and stored in PBS. To visualize dopamine axons in the striatum and dopamine cell bodies in the midbrain, brain sections were incubated with rabbit anti-tyrosine hydroxylase antibodies (TH; AB152, MilliporeSigma, MO) at 4°C overnight and then with fluorescent secondary antibodies (A-11012, Thermo Fisher Scientific, MA) at 4°C overnight. Slices were then mounted in 4',6-diamidino-2-phenylindole (DAPI)-containing anti-fade solution (VECTASHIELD anti-fade mounting medium, H-1200, Vector Laboratories, CA) and imaged with Zeiss Axio Scan Z1 slide scanner fluorescence microscope (Zeiss, Germany).
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