Abnormal accumulation of amyloid-β (Aβ) plaques in the brain is one of the major characteristics of Alzheimer’s disease and there is a possibility that the degree of plaque toxicity is related to the distribution of nanoscale oligomeric aggregates in plaques. Here, by super-resolution fluorescence imaging, we visualized that anti-Aβ oligomer antibodies localized differently to plaques compared to anti-Aβ monomer antibodies and characteristic plaque shapes in brain tissues were classified. These differences in nanoscale distribution were hard to be discerned by conventional fluorescence imaging, implying that super-resolution imaging has the potential to reveal the detailed features of oligomeric aggregates in plaques.
KEYWORDS: Signal detection, Super resolution, Brain, Luminescence, Visualization, Spatial resolution, Signal attenuation, Proteins, Molecules, Microscopy
Abnormal deposition of amyloid-β (Aβ) causes the formation of senile plaques, which is one of the main pathological features of Alzheimer disease. To visualize the plaques at nanometer resolution, single-molecule localization microscopy (SMLM) is promising. It is also essential to develop a method for suppressing autofluorescence especially under high-magnifications used for detecting single molecules. Here, we report a novel method to reduce autofluorescence in mouse brains which is applicable to visualize the structure of Aβ plaques by SMLM. The super-resolution images of Aβ plaques showed fibrous structures that were not able to be discerned by conventional fluorescence imaging.
Single molecule localization microscopy (SMLM) relies on the detection of fluorescence emission from a single fluorophore molecule. Recently, several fluorescent dyes with spontaneous blinking have been reported. Last year, we reported a near-infrared dye with spontaneous blinking and high photostability. Here, we present a new orangefluorescent dye that exhibits spontaneous blinking based on equilibrium between the fluorescent and non-fluorescent forms. We have also ascertained that the new dye can be used to create a super-resolution image of cytoskeletal microtubules. In combination with our near-infrared dye reported last year, the new dye is expected to allow dual-color SMLM.
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