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We counted positive cells in every six sections.
The density of BrdU cells in the GCL was obtained by counting the number of BrdU cells in the GCL of one in every six sections (n = 6 8 mice/group).
Visualization of putative inhibitory neurons was performed with double immunostaining by pairing p75NTR with Pv or 67 kDa glutamic acid decarboxylase (GAD67) antibodies (Table 1) in one in every six sections at the level of the NB.
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One out of every eight sections was processed for ChAT immunocytochemistry, using a mouse monoclonal antibody raised against whole, purified rat brain ChAT-17.
Overall, three out of every five sections have sold out since the pricey club seats went on sale a year ago and the remaining seats began selling in June, Guido said.
One out of every ten sections was selected for each kind of staining.
Since we counted one out of every four sections, we estimate that there are about 252 GFP neurons in the vlPAG per hemisphere in FVB-GFP mice.
The first of every five sections was subjected to Nissl staining of neurons and Klüver-Barrera counterstaining of myelinated nerve fiber tracts.
For the quantitative evaluation of Mvh-positive cells, serial sections of female genital ridges were stained for DNA, Mvh, and phospho-Histone 3. Mvh-positive cells from at least three sections selected by every two sections in at least three embryos (six genital ridges) were scored for each time point.
For the quantitative evaluation of mitotic index, serial sections of female genital ridges were stained for DNA, Mvh, and phospho-Histone 3, Mvh single positive cell, as well as Mvh and phospho-Histone 3 double positive cells were scored from at least three sections selected by every two sections in at least three embryos (six genital ridges) for each time point.
The second of every five sections was immersed in 3% H2O2, 0.05 M Tris, 0.9% NaCl, pH 7.5 for 30 minutes, washed and immersed for 60 minutes in 0.25% Triton ×100, 5% BSA and 0.1 M DL-lysine for α-synuclein immunohistochemistry.
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