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Vol. 15, Issue 3, 957-962, March 2004

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Essay

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

Carolyn A. Larabell ^*  , and Mark A. Le Gros 

^* Department of Anatomy, University of California, San Francisco California 94143; Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720

Submitted July 23, 2003; Revised October 22, 2003; Accepted October 24, 2003
Monitoring Editor: Thomas Pollard

We examined the yeast, Saccharomyces 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 (3-D) 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. Because proteins can be localized in the X-ray microscope using immunogold labeling protocols (Meyer-Ilse et al., 2001. J. Microsc. 201, 395–403), tomography enables 3-D molecular localization. The time required to collect the data for each cell shown here was <15 min and has recently been reduced to 3 min, making it possible to examine numerous yeast and to collect statistically significant high-resolution data. In this video essay, we show examples of 3-D tomographic reconstructions of whole yeast and demonstrate the power of this technology to obtain quantifiable information from whole, hydrated cells.

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

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