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Automated Very Low Magnification Imaging for TEM

Published online by Cambridge University Press:  02 July 2020

C.S. Potter
Affiliation:
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801.
B. Carragher
Affiliation:
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801.
D. Kriegman
Affiliation:
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801.
J. Pulokas
Affiliation:
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801.

Abstract

A typical TEM specimen grid provides approximately a 2×2 mm area that is available for imaging. in order to identify and locate suitable targets on the grid the microscopist must usually inspect the grids at magnifications that allow for only a small area of the grid in the field of view at a time. Systematically searching the grid and mentally keeping track of relative locations presents a challenge for a microscopist especially as the image normally rotates as the magnification is changed. We present an automated technique that creates a very low magnification (VLM) image of the entire available imaging area on the grid. The VLM image can then be used as a reference map for searching the grid at high magnification.

VLM images were obtained at a nominal magnification of 57x using a Philips CM200 TEM equipped with a Gatan CCD camera. The VLM image of the entire 2 mm specimen grid can be created using a mosaic of 49 images, each 512×512 pixels in size. The sampling distance between the images in the 7×7 array is 300μm and the pixel size is 680nm. The images are then automatically tiled and re-sampled to form a final VLM matrix of 4K×4K with a pixel size of 730nm. The resampling factors for the tiling operation include the scale and the relative angle between the camera and goniometer axes. These factors are determined automatically from a calibration process that characterizes the goniometer.

Type
Instrument Automation (Organized by W. Deruijter and C. Potter)
Copyright
Copyright © Microscopy Society of America 2001

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References

1.Pulokas, J., et al., J. Struct. Biol. 128 (2000) 250256.CrossRefGoogle Scholar
2.Carrager, B., et al., J. Struct Biol. 132 (2000) 3345.CrossRefGoogle Scholar
3. This research is supported by the National Science Foundation (DBI-9730056, DBI-9904547) and the National Institutes of Health (GM61939-01).Google Scholar

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