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Author Notes:

Email Address : Alexa L. Mattheyses : mattheyses@emory.edu ; Khalid Salaita :ksalaita@emory.edu

S.M.S. and T.O. conceived the STAR technique and did initial tests.

A.L.M. and K.S. supervised the project. D.R.S. performed all experiments and data analysis.

A.L.M. designed and implemented the theoretical modelling.

All authors contributed to preparing the manuscript.

Subjects:

Research Funding:

K.S. is grateful for support from the NIH through R01-GM097399, the Alfred P. Sloan Research Fellowship and the Camille-Dreyfus Teacher-Scholar Award.

A.L.M. acknowledges support from the NIH through R21-AR066920 and an Emory University Research Committee Grant.

Both K.S. and A.L.M. acknowledge support from the NSF through an IDBR (1353939) grant.

Real-time fluorescence imaging with 20 nm axial resolution

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Journal Title:

Nature Communications

Volume:

Volume 6

Publisher:

, Pages 8307-8307

Type of Work:

Article | Final Publisher PDF

Abstract:

All rights reserved. Measuring the nanoscale organization of protein structures near the plasma membrane of live cells is challenging, especially when the structure is dynamic. Here we present the development of a two-wavelength total internal reflection fluorescence method capable of real-time imaging of cellular structure height with nanometre resolution. The method employs a protein of interest tagged with two different fluorophores and imaged to obtain the ratio of emission in the two channels. We use this approach to visualize the nanoscale organization of microtubules and endocytosis of the epidermal growth factor receptor.

Copyright information:

© 2015, Nature Publishing Group, a division of Macmillan Publishers Limited.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/).

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