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Proceedings Paper

Illuminating cellular structure and function in the early secretory pathway by multispectral 3D imaging in living cells
Author(s): Jens Rietdorf; David J. Stephens; Anthony Squire; Jeremy Simpson; David T. Shima; Jean-Pierre Paccaud; Philippe I.H. Bastiaens; Rainer Pepperkok
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Paper Abstract

Membrane traffic between the endoplasmic reticulum (ER) and the Golgi complex is regulated by two vesicular coat complexes, COPII and COPI. COPII has been implicated in selective packaging of anterograde cargo into coated transport vesicles budding from the ER. COPI-coated vesicles are proposed to mediate recycling of proteins from the Golgi complex to the ER. We have used multi spectral 3D imaging to visualize COPI and COPII behavior simultaneously with various GFP-tagged secretory markers in living cells. This shows that COPII and COPI act sequentially whereby COPI association with anterograde transport complexes is involved in microtubule-based transport and the en route segregation of ER recycling molecules from secretory cargo within TCS in transit to the Golgi complex. We have also investigated the possibility to discriminate spectrally GFP fusion proteins by fluorescence lifetime imaging. This shows that at least two, and possibly up to three GFP fusion proteins can be discriminated and localized in living cells using a single excitation wavelength and a single broad band emission filter.

Paper Details

Date Published: 27 April 2000
PDF: 10 pages
Proc. SPIE 3921, Optical Diagnostics of Living Cells III, (27 April 2000); doi: 10.1117/12.384216
Show Author Affiliations
Jens Rietdorf, European Molecular Biology Lab. (Germany)
David J. Stephens, European Molecular Biology Lab. (Germany)
Anthony Squire, Imperial Cancer Research Fund (United Kingdom)
Jeremy Simpson, European Molecular Biology Lab. (Germany)
David T. Shima, Imperial Cancer Research Fund (United Kingdom)
Jean-Pierre Paccaud, Univ. of Geneva School of Medicine (Switzerland)
Philippe I.H. Bastiaens, Imperial Cancer Research Fund (Netherlands)
Rainer Pepperkok, European Molecular Biology Lab. (Germany)

Published in SPIE Proceedings Vol. 3921:
Optical Diagnostics of Living Cells III
Daniel L. Farkas; Daniel L. Farkas; Robert C. Leif, Editor(s)

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