Mar 03, 2022
TH Densitometry
- 1University of Ottawa
Protocol Citation: Haley Geertsma 2022. TH Densitometry. protocols.io https://dx.doi.org/10.17504/protocols.io.b5u4q6yw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: March 03, 2022
Last Modified: March 03, 2022
Protocol Integer ID: 59004
Abstract
This protocol is used to quantify the density of dopaminergic projections to the striatum.
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
Capture brightfield images of the dorsolateral striatum and convert to greyscale images.
Take 20 intensity measurements of the striatum and 5 intensity measurements of the corpus callosum (to account for background staining).
Average the corpus callosum intensities then subtract that from the striatal intensities then average the normalized striatal intensitied.