FRET imaging of multiple focal planes to analyze the organization and conformation of transferrin-receptor in polarized cells

Horst Wallrabe; Ammasi Periasamy; Margarida Barroso

doi:10.1117/12.810246

25 February 2009 FRET imaging of multiple focal planes to analyze the organization and conformation of transferrin-receptor in polarized cells

Horst Wallrabe, Ammasi Periasamy, Margarida Barroso

Author Affiliations +

Proceedings Volume 7183, Multiphoton Microscopy in the Biomedical Sciences IX; 71832F (2009) https://doi.org/10.1117/12.810246
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

We developed a FRET-based assay to analyze multiple focal planes from z-staks collected using confocal imaging of polarized epithelial MDCK cells. To establish the imaging assay for multiple focal planes, we have used fixed cells, where changes in the organization and conformation of transferrin-receptor complexes between endosomes are expected to be minimized. Therefore, we indeed only see minor changes in E% throughout the endocytic pathway of polarized cells. In the future, we will apply this FRET-based assay to live polarized epithelial cells to investigate the factors involved in the regulation of the organization and conformation of membrane-bound receptors during endocytic trafficking.

Citation Download Citation

Horst Wallrabe, Ammasi Periasamy, and Margarida Barroso "FRET imaging of multiple focal planes to analyze the organization and conformation of transferrin-receptor in polarized cells", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 71832F (25 February 2009); https://doi.org/10.1117/12.810246

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