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Title: Validation of super-resolution 4Pi microscopy, and its application to the study of intra-Golgi transport
Author: Perinetti, Giuseppe
ISNI:       0000 0001 3484 0524
Current Institution: Open University
Date of Award: 2008
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Two problems have hampered the use of light microscopy for structural studies of cellular organelles for a long time: the limited spatial resolution, and the difficulty of obtaining true structural boundaries from complex intensity curves. The advent of modern super-resolution light-microscopy techniques and their combination with objective image segmentation now provide us with the means to bridge the gap between light and electron microscopy in cell biological applications. This study provides the first comparative correlative analysis of threedimensional (3D) structures obtained by 4Pi-microscopy with those of transmission electron microscopy (TEM). The distribution within the cisternae of isolated Golgi stacks of the cargo protein PC3-pEGFP is here mapped in 3D by both 4Pi-microscopy and TEM for a detailed comparative analysis of their imaging capabilities. A high correlation was seen for these structures, demonstrating the particular accuracy of 4Pi-microscopy. Furthermore, for the first time, the transport of a cargo molecule (VSVG-pEGFP) through individual Golgi stacks (labelled by GalT-venusYFP) »s visualized at the 4Pi-microscope super-resolution level.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available