2.9. Immunocytochemistry

Primary hippocampal neurons fixed in 10% buffered formalin were blocked in 10% goat serum for 1 h, then incubated with anti-Bassoon (1:10, Santa Cruz Biotechnology, Dallas, TX, USA), anti-synaptophysin (1:10, Santa Cruz Biotechnology, Dallas, TX, USA), anti-MAP2 (1:100, Cell Signaling Technology, Danvers, MA, USA) antibodies overnight at 4 °C. Cells were washed and incubated with Alexa-fluor 488 antibody or Alexa-568 antibody (1:200 dilution; Invitrogen, Molecular Probes, Carlsbad, CA, USA) for 1 h at room temperature and mounted on glass slides. Images were taken with a Zeiss Axiovert A1 microscope and processed using AxioVision 4.9.