P21 3min Anoxia Conditions*
Brain was removed from a 21 day-old rat following decapitation
and the right hippocampus dissected out. The whole hippocampus was
immersed in a petri dish containing
mixed aldehydes: 2% paraformaldehyde, 6% glutaraldehyde, 1 mM CaCl2,
2 mM MgCl2, at pH 7.4. The dish was placed immediately on a bed of ice in
a microwave oven and microwaved for 60 seconds to speed fixation.
The elapsed time
from death until the start of microwave processing was 3 minutes. The
final temperature of the fixation solution was 36.6 °C.
The intact hippocampus was stored overnight at room temperature
in this fixative and then processed for serial electron microscopy using
standard procedures.
*All procedures follow
NIH Guidelines for the humane care and use of laboratory animals.
