1 research outputs found
Nanoscale Fluorescence Imaging of Single Amyloid Fibrils
Amyloid formation is implicated in a variety of human diseases.
It is important to perform high-resolution optical imaging of individual
amyloid fibrils to delineate the structural basis of supramolecular
protein assembly. However, amyloid fibrils do not lend themselves
to the conventional microscopic resolution, which is hindered by the
diffraction limit. Here we show super-resolution fluorescence imaging
of fluorescently stained amyloid fibrils derived from disease-associated
human β<sub>2</sub>-microglobulin using near-field scanning
fluorescence microscopy. Using this technique, we were able to resolve
the fibrils that were spatially separated by ∼75 nm. We have
also been able to interrogate individual fibrils in a fibril-by-fibril
manner by simultaneously monitoring both nanoscale topography and
fluorescence brightness along the length of the fibrils. This method
holds promise to detect conformational distributions and heterogeneity
that are believed to correlate with the supramolecular packing of
misfolded proteins within the fibrils in a diverse conformationally
enciphered prion strains and amyloid polymorphs