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Real-time fluorescence imaging with 20?nm axial resolution.


ABSTRACT: 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.

SUBMITTER: Stabley DR 

PROVIDER: S-EPMC4595625 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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Real-time fluorescence imaging with 20 nm axial resolution.

Stabley Daniel R DR   Oh Thomas T   Simon Sanford M SM   Mattheyses Alexa L AL   Salaita Khalid K  

Nature communications 20150922


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 appro  ...[more]

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