Fig. 7

CLEM identification of senile plaques and neurofibrillary tangles on the same EM section using dual immunofluorescence staining. Two immediate ultrathin sections, 120 nm and 70 nm thick, were obtained from embedded AD brain tissue. The 120 nm section underwent immunofluorescence staining with primary antibodies against Tau (chicken-derived) and Aβ (rabbit-derived), followed by secondary antibodies conjugated with Alexa Fluor™ 488 goat anti-chicken (green signal) and Alexa Fluor™ 594 goat anti-rabbit (red signal). The 70 nm section was subjected to UA and lead staining to generate EM images. CLEM was used to align the fluorescence LM signals with the ultrastructural EM details, allowing visualization of senile plaques and neurofibrillary tangles in the EM image. Framed regions in the left images are magnified in closeup1, and further detailed enlargements are presented in closeup2. Scale bars are included in all images