Immunohistochemistry for Identifying the Locus Coeruleus Region in Mouse Brain Sections

The locus coeruleus regions, located bilaterally in the mouse brainstem, contain norepinephrine-producing neurons and regulate various regions across the brain.

To detect these locus coeruleus regions, begin with a glass slide carrying the fixed, cryopreserved mouse brainstem sections.

Wash the slide with a buffer to remove fixative traces.

Add a buffer containing detergent and incubate to permeabilize the membranes, then wash.

Introduce primary antibodies specific to dopamine β-hydroxylase enzyme, which converts dopamine to norepinephrine, a neurotransmitter.

Incubate to allow the antibodies to interact with dopamine β-hydroxylase.

Wash to remove unbound antibodies.

Then, add green fluorophore-coupled secondary antibodies and incubate in the dark.

The secondary antibodies bind to the primary antibodies and then wash.

Add mounting medium to the brain sections, then cover and dry.

Observe the slides under a fluorescence microscope.

The green fluorescence representing the labeled norepinephrine-producing neurons in the brain sections indicates the presence of locus coeruleus regions.