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MBC in Press, published online ahead of print December 29, 2003
Mol. Biol. Cell 10.1091/mbc.E03-07-0522

A more recent version of this article appeared on March 1, 2004
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Submitted on July 23, 2003
Revised on October 22, 2003
Accepted on October 24, 2003

X-ray Tomography Generates 3-D Reconstructions of the Yeast, Saccharomyces cerevisiae, at 60 nm Resolution

Carolyn A. Larabell1* and Mark A. Le Gros2

1 Department of Anatomy, University of California, San Francisco CA, 94143; Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720
2 Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720

* Corresponding author. E-mail address: larabel{at}itsa.ucsf.edu.

We examined the yeast, S. cerevisiae, using x-ray tomography and demonstrate unique views of the internal structural organization of these cells at 60 nm resolution. Cryo x-ray tomography is a new imaging technique that generates three-dimensional information of whole cells. In the energy range of x-rays used to examine cells, organic material absorbs approximately an order of magnitude more strongly than water. This produces a quantifiable natural contrast in fully hydrated cells and eliminates the need for chemical fixatives or contrast enhancement reagents to visualize cellular structures. Since proteins can be localized in the x-ray microscope using immunogold labeling protocols (Meyer-Ilse et al., 2001), tomography enables three-dimensional molecular localization. The time required to collect the data for each cell shown here was <15 min and has recently been reduced to three minutes, making it possible to examine numerous yeast and to collect statistically significant high-resolution data. In this video essay, we show examples of three-dimensional tomographic reconstructions of whole yeast and demonstrate the power of this technology to obtain quantifiable information from whole, hydrated cells.




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