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Automatic Recording of Low Dose Images with Slow-Scan CCD Cameras

Published online by Cambridge University Press:  02 July 2020

E. Zellmann
Affiliation:
LEO Elektronenmikroskopie GmbH, Carl-Zeiss-Str. 56, D-73446Oberkochen/, Germany
W. Probst
Affiliation:
LEO Elektronenmikroskopie GmbH, Carl-Zeiss-Str. 56, D-73446Oberkochen/, Germany
V. Seybold
Affiliation:
LEO Elektronenmikroskopie GmbH, Carl-Zeiss-Str. 56, D-73446Oberkochen/, Germany
S. A. Hiller
Affiliation:
LEO Elektronenmikroskopie GmbH, Carl-Zeiss-Str. 56, D-73446Oberkochen/, Germany
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Extract

Recording of images under low dose conditions is a rather critical operation including many intermediate steps. Moreover, there is not a single fully agreed protocol concerning low dose operation but several competing ones potentially including individual variations. In any case, however, there should be a very well defined sequence of steps which must not allow for any mistake. Therefore tailored individual automation is the only way to obtain reliable results with maximum throughput at the same time.

Such automation requires the functional integration of a fully remote controllable digital TEM, a Slow-Scan camera with a high sensitivity, and an acquisition/remote control software providing macro capabilities dedicated to different logical cameras. These requirements are met by LEO (EF)TEMs, by the ProScan SSCCD 14 bit camera, and the analySIS software 3.0.

Three different logical cameras have to be configured in the SIS image processing software. The first camera is defined for searching for suitable specimen areas. It operates with 5 MHz and 8 bit image depth providing 1k×1k field size and resolution. The exposure time should be as short as possible, e.g. 50 ms for each frame. Via a preprocessing macro image brightness and EM magnification are set to the appropriate values, the desired shutter is activated, and the viewing screen is lifted for the exposure. A second postprocessing macro blanks the beam and does additional settings when the camera is stopped.

Type
Cryotechniques, Immunocytochemistry, and Electron Microscopy
Copyright
Copyright © Microscopy Society of America

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