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Localization microscopy for the study of amyloid fibril formation, Pinotsi D, Kaminski-Schierle GS, Rees EJ, Kaminski CF, Proc.SPIE 88150G, Nanoimaging and Nanospectroscopy Proc.SPIE 88150G, Nanoimaging and Nanospectroscopy, 88150G (September 20, 2013)

DOI: 10.1117/12.2022589 | pdf


Abstract

Super-resolution microscopy has emerged as a powerful and non-invasive tool for the study of molecular processes both in vitro, but also as they occur in live cells. Here we present the application of direct stochastic optical reconstruction microscopy (dSTORM), a super-resolution technique based on single molecule localization, to determine the morphology of protein aggregates and of small extra- and intracellular structures. The technique reveals details down to 20 nm providing information on scales much smaller than the wavelength of the probing light. We use dSTORM in the study of amyloid fibril self-assembly processes associated with neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases. We show that the aggregation process can be followed kinetically and observe the emergence of amyloid structures in time as they occur in vitro. As an all optical technique, there is translation potential from studies in vitro to in vivo applications.

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