Research Article: Imaging Transient Blood Vessel Fusion Events in Zebrafish by Correlative Volume Electron Microscopy

Date Published: November 6, 2009

Publisher: Public Library of Science

Author(s): Hannah E. J. Armer, Giovanni Mariggi, Ken M. Y. Png, Christel Genoud, Alexander G. Monteith, Andrew J. Bushby, Holger Gerhardt, Lucy M. Collinson, Amy S. Gladfelter.

Abstract: The study of biological processes has become increasingly reliant on obtaining high-resolution spatial and temporal data through imaging techniques. As researchers demand molecular resolution of cellular events in the context of whole organisms, correlation of non-invasive live-organism imaging with electron microscopy in complex three-dimensional samples becomes critical. The developing blood vessels of vertebrates form a highly complex network which cannot be imaged at high resolution using traditional methods. Here we show that the point of fusion between growing blood vessels of transgenic zebrafish, identified in live confocal microscopy, can subsequently be traced through the structure of the organism using Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) and Serial Block Face/Scanning Electron Microscopy (SBF/SEM). The resulting data give unprecedented microanatomical detail of the zebrafish and, for the first time, allow visualization of the ultrastructure of a time-limited biological event within the context of a whole organism.

Partial Text: Progress has been made in the field of high resolution electron microscopy to the point where structural determination of single molecules is becoming routine. However, the challenge of obtaining high resolution structural data from bulk samples of cells, tissues and whole organisms remains largely unexplored.

We have developed a method combining live fluorescence imaging with automated volume EM to produce unprecedented three-dimensional structural and functional detail of a transient biological process in vivo. This technique could find wide application in the fields of cell biology, vascular biology, developmental biology, pathology and neuroscience.



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