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Cell numbers and vascular effects were assessed by immunohistochemistry using several (typically 6) coronal sections taken between +21 mm and +31 mm relative to Ear Bar Zero (EBZ).
The ear bar was inserted into the ear canal of the mouse and secured.
The signal was sent to the speaker connected to the ear bar, which was calibrated with the microphone.
The center of the chamber was placed at 15.0 17.5 mm anterior to the ear bar position.
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Once dissection was complete, the preparation was placed in a supine position in a specially designed plexiglass chamber and secured with ear bars.
The animals were placed in a stereotaxic frame and the head fixed using ear bars and a snout clamp.
The animal was placed in a stereotaxic frame with the head held in a fixed position by ear bars.
Mice were anaesthetized using Chloral hydrate and placed into a stereotactic frame with nose and ear bars specially adapted for mice.
The animals were transferred to a stereotaxic frame to hold the head in a fixed horizontal position using ear bars and a snout clamp.
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