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A total of 550,000 particles (window size 144 × 144) were automatically picked from 2,100 micrographs.
Protein spots of interest were automatically picked from the 2D gel with the Ettan Spot Picker, subject to tryptic digestion and MS/MS analysis.
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For three-dimensional (3D) reconstruction of MG23 structures, particle images for the large bowl-shaped and asymmetric small structures were automatically picked up from the electron micrographs.
Automatic structure calculation was conducted using, in addition to the information from the manual calculation, automatically picked peak lists from the CHHC, PAR, PDSD, and two PAIN spectra (Table S2) for iterative assignment by CYANA (Table S2).
For instance, instead of having to manually set the camera for each mode of shooting (such as normal or macro), a feature called Intelligent IS automatically picks from six modes: normal, macro, panning dynamic, powered and tripod, to avoid blurred photos.
Using this selection criterion, 196 micrographs were selected, from which a total of 87,066 particles were automatically picked using FindEM (Roseman, 2004).
Automatically picked particles were manually verified using a correspondence analysis based method (Shaikh et al., 2008b).
"A bag left unattended would be automatically picked up by the system".
Their method was highly accurate, even when automatically picked peaks were used as the inputs.
The automatically picked particles were then extracted into 256 by 256 boxes, CTF corrected, and cropped as before.
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