Axioimager z1

Differentiated human neurons were fixed and stained with antibodies directed against neurofilament (1:1000; Sigma, Germany), βIII tubulin (1:500; Sigma, Germany), microtubule-associated protein 2 (MAP2; 1:200; Millipore, Germany), vesicular glutamate transporter 1 (vGLUT1; 1:300; Synaptic Systems, Germany), gamma aminobutyric acid (GABA; 1:500; Sigma, Germany), tyrosine hydroxylase (TH; 1:1000; Sigma, Germany), choline acetyltransferase (ChAT; 1:100; Millipore, Germany), glial fibrillary acidic protein (GFAP; 1:500; DAKO, Denmark) or the ionized calcium binding adapter molecule 1 (Iba1; 1:500; Wako, Japan) followed by the corresponding Alexa488-conjugated antibodies (1:500; Invitrogen, Germany). Neurons were counterstained with an antibody directed against human neuronal nuclei (NeuN; 1:50; Millipore, Germany) or βIII tubulin (1:500) followed by the corresponding Cy3-conjugated secondary antibody (1:500; Dianova, Germany). Nuclei of cells were subsequently labeled with 4´,6-diamidino-2-phenylindole (DAPI; 1:10,000; Sigma, Germany). Images were taken by confocal laser scanning microscopy (Fluoview 1000, Olympus) or fluorescence microscope (AxioImager.Z1).