The development of methods able to access a precise molecular counting of protein copy numbers is essential, clearing the way to address several biological questions using super-resolution techniques such as stochastic optical reconstruction microscopy (STORM). A suitable calibration method represents the best way to address the challenges of molecular counting using super-resolution [3, 4] since small organic fluorophores used in conjunction with immunofluorescence need to be characterized for quantitative imaging.Here we propose to use DNA origami as a standard and we show that this calibration method, besides quantifying the average protein copy number in a cell, allows determining the abundance of various oligomeric states [5]. In particular, we show that quantitative approaches allow quantitative studies of the stoichiometry of membrane proteins …

Quantitative Super-Resolution Microscopy using DNA Origami

Cella Zanacchi, Francesca;Diaspro, Alberto;
2018

Abstract

The development of methods able to access a precise molecular counting of protein copy numbers is essential, clearing the way to address several biological questions using super-resolution techniques such as stochastic optical reconstruction microscopy (STORM). A suitable calibration method represents the best way to address the challenges of molecular counting using super-resolution [3, 4] since small organic fluorophores used in conjunction with immunofluorescence need to be characterized for quantitative imaging.Here we propose to use DNA origami as a standard and we show that this calibration method, besides quantifying the average protein copy number in a cell, allows determining the abundance of various oligomeric states [5]. In particular, we show that quantitative approaches allow quantitative studies of the stoichiometry of membrane proteins …
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11567/892727
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