Fig. 1

In-situ immunohistochemical detection of nuclear aSyn. Example micrograph images from the cingulate cortex of control (con) and dementia with Lewy body (DLB) cases of phospho-serine 129 positive aSyn (pS129) mouse IgG2 (a), pan-aSyn N-terminal directed Syn-303 (b), pan-aSyn non-amyloid component directed Syn1 (c) and NeuN and pS129 rabbit EP1536Y (d) immunoreactivity. All sections were pre-treated with EDTA + formic acid antigen retrieval, nuclei in sections were counterstained with DAPI. Images captured via 40× (a–c, i), 60 × objective lens (d, i) with wide-field fluorescence microscope and 20 × objective lens with 6 × digital zoom (c, iii) or 63× objective lens with 3 × digital zoom (d, ii) with confocal microscope. Expanded area inserts (a–c, ii, dotted line boxes in i are shown for aSyn immunoreactive alone and in combination with DAPI nuclear stain, where the nuclear outline is highlighted (dotted outline). Note frequent detection of intranuclear aSyn immunoreactivity, examples highlighted by arrow heads, in d NeuN-negative nuclei positive for aSyn are further denotes with an asterisk. Scale bar in i = 10 µm and ii and iii = 5 µm