Stereology
Haley Geertsma
Published: 2022-03-06 DOI: 10.17504/protocols.io.b5ynq7ve
Abstract
This protocol is used to quantify the number of TH+ cells in the substantia nigra pars compacta.
Steps
1.
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
2.
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).
