Mar 05, 2022
Stereology
- 1University of Ottawa
Protocol Citation: Haley Geertsma 2022. Stereology. protocols.io https://dx.doi.org/10.17504/protocols.io.b5ynq7ve
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 05, 2022
Last Modified: March 05, 2022
Protocol Integer ID: 59118
Abstract
This protocol is used to quantify the number of TH+ cells in the substantia nigra pars compacta.
Take 8 cryosections cut at 40μm to equally sample the substantia nigra, pars compacta (SNc) and perform diaminobenzidine staining with a tyrosine hydroxylase antibody.
Note: I take every 6th section from Bregma -2.54mm to -3.88mm
2d
Use StereoInvestigator software to count all TH+ cells. For this, randomly sample the SNc with a 150μm x 150μm grid and apply an optical fractionator consisting of a 75μm x 75μm square. Calculate the mean section thickness during counting with a frequency of 10 frames (~3 measurements per hemisphere).
1h