Cleaved caspase-3 staining ruled out that this was due to increased cell death in SVZ of female mutants (Fig. S2D, E).
Also, while showing some minor fresh hemorrhages as a result of the surgical retrieval of the mesh bag, the H&E staining ruled out major local tissue injury.
This finding, together with the results of the Ki67 staining, ruled out the possibility that the increased number of tissue macrophages was due to in situ proliferation, as noted during inflammation (Davies et al., 2013).
To confirm the presence of Aβ in the choroidal vasculature, another method of immunohistochemistry was performed using a colorimetric staining to rule out all background staining that might result in false positive staining.
Because of the resemblance of the wing lesions to those typically seen in white nose syndrome infection, which is caused by the fungus Pseudogymnoascus destructans, we applied periodic acid Schiff staining to rule out fungal infection.
All specimens were stained with Ziehl-Nielson and Gomori methenamine silver stains to rule out infectious granulomas caused by acid-fast organisms and fungi, respectively.
Non-specific fluorescence staining was ruled out using non-specific isotypic antibodies (data not shown).
FG, FO and JW carried out the secondary immunohistochemical staining evaluation to rule out interindividual observer discrepancies.
However, for clinical purposes, it may be of use to use polySia-NCAM antibodies in immunohistochemical stainings of astrocytomas, since positive staining very likely rules out the possiblity of pilocytic astrocytomas.
Unspecific staining can be ruled out by evaluating normal BM cells in the sample, which are supposed to be completely negative for GD2 staining.
