Generation of Medial Thalamus-Anterior Cingulate Cortex Slices from Mouse Brains
Trascrizione
Expose the skull of the animal, and trim off the remaining muscle. Next, peel away the dorsal surface of the skull using rongeurs and trim away the sides. Subsequently, using a spatula, cut the olfactory bulbs and nerve connections along the ventral surface of the brain before removing it.
Quickly transfer the brain to a beaker filled with ice-cold oxygenated ACSF. To prepare slices containing the MT-ACC pathway, make a sagittal cut 2 millimeters lateral to the midline in each hemisphere to display the subcortical anatomy.
Then, make an angled crosscut parallel to the visible fiber tract in the striatum. Make the second angled cross-cut from the connection between the cerebellum and visual cortex, to the midpoint between the anterior commissure and the optic tract that are ventral and parallel to the thalamus cingulate pathway. After that, attach the brain block to an angular plate with a cyanoacrylate adhesive.
Make a cut just above the turning point of the pathway. After that, unfold the plate, flatten it, and glue it onto the chamber stage of a vibratome. Subsequently, section the brain slices at 500 micrometers thickness and keep them in ice-cold oxygenated ACSF. Transfer a slice to the recording chamber at 32 degrees Celsius with continuous perfusion of oxygenated ACSF for one hour.
