TH Densitometry
Haley Geertsma
Published: 2022-03-03 DOI: 10.17504/protocols.io.b5u4q6yw
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Abstract
This protocol is used to quantify the density of dopaminergic projections to the striatum.
Steps
1.
Take 4 cryosections to equally sample the striatum and perform diaminobenzidine staining with a tyrosine hydroxylase antibody.
Note: I take every 12th section from Bregma 1.18mm to -0.22mm
2.
Capture brightfield images of the dorsolateral striatum and convert to greyscale images.
3.
Take 20 intensity measurements of the striatum and 5 intensity measurements of the corpus callosum (to account for background staining).
4.
Average the corpus callosum intensities then subtract that from the striatal intensities then average the normalized striatal intensitied.
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