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Applications in Stimulated Emission Depletion Microscopy: Localization of a Protein Toxin in the Endoplasmic Reticulum Following Retrograde Transport

Published online by Cambridge University Press:  02 November 2016

Cristina Herrera ,
Nicholas J. Mantis  and
Richard Cole
Cristina Herrera
Affiliation:
Wadsworth Center, Division of Infectious Disease, New York State Department of Health, Albany, NY 12208, USA Department of Biomedical Sciences, University at Albany School of Public Health, Albany, NY 12201, USA
Nicholas J. Mantis
Affiliation:
Wadsworth Center, Division of Infectious Disease, New York State Department of Health, Albany, NY 12208, USA Department of Biomedical Sciences, University at Albany School of Public Health, Albany, NY 12201, USA
Richard Cole*
Affiliation:
Department of Biomedical Sciences, University at Albany School of Public Health, Albany, NY 12201, USA Wadsworth Center, Division of Translational Medicine, New York State Department of Health, Albany, NY 12201, USA
*
*Corresponding author.richard.cole@health.ny.gov
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Abstract

Retrograde transport is a process in which proteins are trafficked from the plasma membrane and endosomes to biosynthetic and secretory organelles, namely the Golgi apparatus and endoplasmic reticulum (ER). A number of plant and bacterial toxins, including cholera toxin and ricin toxin, exploit retrograde transport to gain entry into host cells, although the specifics of this process have remained difficult to probe by laser scanning confocal microscopy (LSCM). Here we demonstrate the use of super-resolution and live-cell imaging [stimulated emission depletion (STED)] to visualize exogenously applied ricin toxin within the ER. The improved resolution obtained by STED, as compared with LSCM (0.09 versus 0.19 μm), provides a more accurate determination of the amount of ricin that had trafficked to the ER.

Keywords

stimulated emission depletionendoplasmic reticulumlive-cell imagingricin
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 22 , Issue 6 , December 2016 , pp. 1113 - 1119
Copyright
© Microscopy Society of America 2016 

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