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Improving Signal to Noise in Labeled Biological Specimens Using Energy-Filtered TEM of Sections with a Drift Correction Strategy and a Direct Detection Device

Published online by Cambridge University Press:  19 March 2014

Ranjan Ramachandra ,
James C. Bouwer ,
Mason R. Mackey ,
Eric Bushong ,
Steven T. Peltier ,
Nguyen-Huu Xuong  and
Mark H. Ellisman
Ranjan Ramachandra*
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
James C. Bouwer
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
Mason R. Mackey
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
Eric Bushong
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
Steven T. Peltier
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
Nguyen-Huu Xuong
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
Mark H. Ellisman
Affiliation:
Center for Research in Biological Systems, National Center for Microscopy and Imaging Research, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
*
*Corresponding author. raramachandra@ucsd.edu
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Abstract

Energy filtered transmission electron microscopy techniques are regularly used to build elemental maps of spatially distributed nanoparticles in materials and biological specimens. When working with thick biological sections, electron energy loss spectroscopy techniques involving core-loss electrons often require exposures exceeding several minutes to provide sufficient signal to noise. Image quality with these long exposures is often compromised by specimen drift, which results in blurring and reduced resolution. To mitigate drift artifacts, a series of short exposure images can be acquired, aligned, and merged to form a single image. For samples where the target elements have extremely low signal yields, the use of charge coupled device (CCD)-based detectors for this purpose can be problematic. At short acquisition times, the images produced by CCDs can be noisy and may contain fixed pattern artifacts that impact subsequent correlative alignment. Here we report on the use of direct electron detection devices (DDD’s) to increase the signal to noise as compared with CCD’s. A 3× improvement in signal is reported with a DDD versus a comparably formatted CCD, with equivalent dose on each detector. With the fast rolling-readout design of the DDD, the duty cycle provides a major benefit, as there is no dead time between successive frames.

Keywords

EFTEMdirect detection deviceDDDCCDDE-12drift correctionendosomesastrocytes
Type
EDGE Special Issue
Information
Microscopy and Microanalysis , Volume 20 , Issue 3 , June 2014 , pp. 706 - 714
Copyright
© Microscopy Society of America 2014 

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