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For double immunofluorescence, consecutive staining by different primary and secondary antibodies was performed.
The majority of NESP55 labelling was located in beta-cells, as evident from consecutive staining for NESP55, CgA and insulin.
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CSF-specific oligoclonal bands (OCB) were determined by isoelectric focusing in polyacrylamide gels with consecutive silver staining.
Islet conformation was studied by confocal analysis in consecutive slices stained for insulin and glucagon.
High-resolution histological images from two consecutive myelin stained sections (spaced at 200 µm) were assembled in separate layers in the Adobe Illustrator file used for image registration (see registration section above).
Tissue distribution of MCP-1 and CCR2 was roughly verified by comparison with consecutive sections stained with hematoxylin-eosin (H&E).
Simultaneous and consecutive double-staining using Prussian blue and CD68 immunostaining were performed for co-localization of SPIONs with macrophages.
The presence of grade-3 lymphoid aggregates, as defined previously [ 12- 15], was assessed on consecutive sections stained for CD3 and CD20.
Dopaminergic neuronal cell bodies from five consecutive sections stained with TH at the comparable SNpc region from three individual mice in each group were counted under blinded conditions by two different researchers independently.
Moreover, consecutive DAB stained slides of CD163 positive macrophages and FcγR positive cells showed that the amount of FcγRI, II and III expression was significantly correlated with CD163 expression in the RA synovia (FcγRI, r = 0.65, p = 0.012; FcγRII, r = 0.85, p = 0.0001; FcγRIII, r = 0.75, p = 0.002), which confirms the results of the immunofluorescence stainings.
One pair includes two images from consecutive sections stained respectively for hematoxylin and eosin (H&E) and antibodies directed against p63 and Alpha-methylacyl-CoA racemase (AMACR), a combination of proteins used in routine clinical diagnostics to identify basal cells and high grade prostate intraepithelial neoplasia (HGPIN /PCa cells, respectively.
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