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J Struct Biol. 2018 Aug;203(2):90-93. doi: 10.1016/j.jsb.2018.03.007. Epub 2018 Mar 15.

Blind estimation of DED camera gain in Electron Microscopy.

Author information

1
Centro Nac. Biotecnología (CSIC), c/Darwin, 3, 28049 Cantoblanco, Madrid, Spain; Univ. San Pablo - CEU, Campus Urb. Montepríncipe, 28668 Boadilla del Monte, Madrid, Spain. Electronic address: coss@cnb.csic.es.
2
Univ. San Pablo - CEU, Campus Urb. Montepríncipe, 28668 Boadilla del Monte, Madrid, Spain.
3
Centro Nac. Biotecnología (CSIC), c/Darwin, 3, 28049 Cantoblanco, Madrid, Spain.
4
Univ. Autónoma de Madrid, 28049 Cantoblanco, Madrid, Spain.

Abstract

The introduction of Direct Electron Detector (DED) videos in the Electron Microscope field has boosted Single Particle Analysis to a point in which it is currently considered to be a key technique in Structural Biology. In this article we introduce an approach to estimate the DED camera gain at each pixel from the movies themselves. This gain is needed to have the set of recorded frames into a coherent gray level range, homogeneous over the whole image. The algorithm does not need any other input than the DED movie itself, being capable of providing an estimate of the camera gain image, helping to identify dead pixels and cases of incorrectly calibrated cameras. We propose the algorithm to be used either to validate the experimentally acquired gain image (for instance, to follow its possible change over time) or to verify that there is no residual gain image after experimentally correcting for the camera gain. We show results for a number of DED camera models currently in use (DE, Falcon II, Falcon 3, and K2).

KEYWORDS:

Direct detectors; Electron microscopy; Single particle analysis

PMID:
29551714
DOI:
10.1016/j.jsb.2018.03.007

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