Stereological Quantification of Dopaminergic Neurons

Stereological countings were performed on 30 μm coronal serial sections analyzing every sixth section for the SNpc and VTA. This was done using an oil immersion 63x objective, a counting frame of 50 x 50 μm, and a grid size of 100 x 100 μm, as described previously [27 (link)] using the optical fractionator method of the StereoInvestigator software 8.0 (MicroBrightField). To quantify lacZ positive and TH positive cells (Fig 1) we counted at least 2 representative 30 μm serial coronal sections. Venus positive and TH positive cells (Fig 2) were quantified in at least 3 representative 50 μm serial sagittal sections per mouse (every forth section) by applying a binary mask and the “Analyze Particles” function in ImageJ (NIH).