Biology Reference
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Coated part of the filament
0mm
10mm
Nylon monofilament
1tick = 1mm
Fig. 1
Thermofusible homemade glue-coated filament of 0.380 mm diameter
and 2.5 mm length
2. Perform a medial incision in the scalp with scalpel after injection
of a drop of bupivacaine.
3. Dissect the temporalis muscle to expose the area of interest
(coordinates 1.5 mm posterior, 5.5 mm lateral to the bregma).
Refine the parietal bone with a saline-cooled dental drill on a
2 mm circle diameter.
4. Position the calibrated LDF probe on the thinned parietal
bone. The LDF probe holder is positioned on the stereotaxic
frame and so even when this one is tilted to induce ischemia,
the LDF probe can't move.
5. Relative CBF (rCBF) is then recorded before and after the
introduction of the filament into the carotid artery.
During the whole procedure, rectal temperature, blood pressure,
heart rate, end tidal CO
2
and blood gazes are monitored, especially
when the experimentation is of a long duration as it is well know
that the evolution of the ischemic lesion is influenced by those
parameters especially the temperature. Here, we describe a full
monitoring of physiological parameters in the case of the animal is
maintained anesthetized during hours following MCAO. This
involves artificial ventilation and invasive measurement of blood
pressure, heart rate and blood samples collection.
3.1.3 Physiological
Parameters Monitoring
1. Rats are anesthetized with isoflurane (5%) in a mixture of 30%
oxygen and 70% nitrous oxide during 5 min in a specific box
under the hood.
2. Rats are intubated and artificially ventilated. If possible, con-
tinuously monitor end tidal CO
2
.
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