Perfusion Conditions*
The brain was fixed in situ via intravascular perfusion with mixed aldehydes
containing 6% glutaraldehyde, 2% paraformaldehyde, 100 mM Cacodylate buffer,
1mM CaCl2, 2 mM MgCl2, pH 7.4, 40-45 °C, and 4 psi pressure under deep pentobarbital
anesthesia (80 mg/Kg). The brain was stored overnight in the fixative at room temperature
and then processed for serial EM using standard procedures.
*All procedures follow
NIH Guidelines for the humane care and use of laboratory animals.
