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Automated acquisition of electron microscopic random conical tilt sets Shawn Q. Zheng a,b
 

Summary: Automated acquisition of electron microscopic random conical tilt sets
Shawn Q. Zheng a,b
, Justin M. Kollman a,b
, Michael B. Braunfeld a,b
, John W. Sedat b
,
David A. Agard a,b,*
a
The Howard Hughes Medical Institute, University of California, San Francisco, CA 94158-2517, USA
b
The W.M. Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco,
CA 94158-2517, USA
Received 28 September 2006; received in revised form 23 October 2006; accepted 26 October 2006
Available online 3 November 2006
Abstract
Single particle reconstruction using the random conical tilt data collection geometry is a robust method for the initial determination of
macromolecular structures by electron microscopy. Unfortunately, the broad adoption of this powerful approach has been limited by the
practical challenges inherent in manual data collection of the required pairs of matching high and low tilt images (typically 60 and 0).
The microscopist is obliged to keep the imaging area centered during tilting as well as to maintain accurate focus in the tilted image while
minimizing the overall electron dose, a challenging and time consuming process. To help solve these problems, we have developed an

  

Source: Agard, David - Department of Biochemistry and Biophysics, University of California at San Francisco

 

Collections: Biotechnology; Biology and Medicine