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Automated CCD Data Collection and 3D Reconstruction of Aleutian Mink Disease Parvovirus

Published online by Cambridge University Press:  02 July 2020

N.H. Olson ,
P.R. Chipman ,
M.E. Bloom ,
R McKenna ,
M. Agbandje-McKenna ,
T.F. Booth  and
T.S. Baker
N.H. Olson
Affiliation:
Department of Biological Sciences, Purdue University, West Lafayette, IN47907
P.R. Chipman
Affiliation:
Department of Biological Sciences, Purdue University, West Lafayette, IN47907
M.E. Bloom
Affiliation:
Laboratory of Persistent Viral Disease, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Disease, Hamilton, MT59840
R McKenna
Affiliation:
Department of Biological Sciences, University of Warwick, CoventryCV4 7AL, England
M. Agbandje-McKenna
Affiliation:
Department of Biological Sciences, University of Warwick, CoventryCV4 7AL, England
T.F. Booth
Affiliation:
NERC Institute of Virology & Environmental Microbiology, Oxford, 0X1 3SR, England
T.S. Baker
Affiliation:
Department of Biological Sciences, Purdue University, West Lafayette, IN47907
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Extract

Acquisition of CCD image data from frozen-hydrated specimens provides several advantages compared with the use of photographic film. CCD cameras possess excellent linearity, sensitivity and dynamic range. The CCD also allows one to pre-check the quality of a cryo sample much easier than with a TV rate camera or on film. However, the limited field of view (IK2 pixels) of the CCD makes data collection difficult. Gatan, Inc. has produced a LowDose software package, in conjunction with their DigitalMicrograph™ environment, that automates this process and allows for efficient collection of larger amounts of data. This software, along with our own image analysis software, has facilitated the computation of a three-dimensional reconstruction (3D) from images of baculo-expressed, VP2 particles of Aleutian mink disease parvovirus (ADV).

ADV has a capsid (∼28.5 nm diam.) that contains two proteins (VP1-2) of which ∼90% is VP2. ADV infections lead to plasmacytosis in mink and are a significant problem worldwide.

Type
Computational Advances and Enabling Technologies for 3D Microscopies in Biology
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1117 - 1118
Copyright
Copyright © Microscopy Society of America 1997

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References

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