Quantifying Dopaminergic Neurons in Mouse Midbrain

The number of dopaminergic neurons was measured by detecting tyrosine hydroxylase (TH) expression in midbrain of mouse. In brief, the midbrain of each mouse was cut into sections to a thickness of 5 μm. The sections were washed using Tris-buffered saline (TBS) and then incubated with TBS+ (0.3% Triton-X100 and 2% bovine serum albumin in TBS) for 45 min. Thereafter, primary antibody against TH (1:500, Boster, Wuhan, China) was used to incubate the sections for 48 h. After being washed by TBS, the sections were incubated with FITC-conjugated secondary antibody (1:500, Boster, Wuhan, China) for 3 h. The TH immunopositive dopaminergic neurons were observed under fluorescence microscope with red fluorescence as positive TH expression signals [41 (link)].