Intracellular Recording In Vivo and Patch-Clamp Recording on Brain Slices - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 106

8. Histological procedures:
At the end of the experiment, the animal is given a large dose
of anesthesia, perfused transcardially with 0.01 M phosphate-
buffered saline (PBS) followed by 4% paraformaldehyde in the
same buffer. The brain is removed and stored in fi xative over-
night. Coronal vibratome sections are cut at 50-mm thickness,
incubated in 0.1% peroxidase-conjugated avidin-D (Vector
Lab) in 0.01 M potassium PBS (KPBS, pH 7.4) with 0.5%
Triton X-100 overnight at room temperature. After detection
of peroxidase activity with 3¢,3¢-diaminobenzidine (DAB),
sections are examined in KPBS. Each recovered neuron is
identifi ed with the recording chart (Fig. 4 ) and matched with
the electrophysiological data. Those sections containing labeled
neurons and stimulation sites are mounted on gelatin-coated
slides and counterstained with cresyl violet for light micros-
copy. Figure 5 shows a neuron recovered after intracellular
staining.
Fig. 5. An example of CA1 neurons in hippocampus after intracellular staining. The section
is counterstained with cresyl violet.