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J Struct Biol. 2003 Jul;143(1):24-32.

Focus gradient correction applied to tilt series image data used in electron tomography.

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  • 1Institute of Molecular Biophysics, Florida State University, Mail Code 4380, Tallahassee, FL 32306-4380, USA.


The resolution in 3D reconstructions from tilt series is limited to the information below the first zero of the contrast transfer function unless the signal is corrected computationally. The restoration is usually based on the assumption of a linear space-invariant system and a linear relationship between object mass density and observed image contrast. The space-invariant model is no longer valid when applied to tilted micrographs because the defocus varies in a direction perpendicular to the tilt axis and with it the shape of the associated point spread function. In this paper, a method is presented for determining the defocus gradient in thin specimens such as sections and 2D crystals, and for restoration of the images subsequently used for 3D reconstruction. The alignment procedure for 3D reconstruction includes area matching and tilt geometry refinement. A map with limited resolution computed from uncorrected micrographs is compared to a volume computed from corrected micrographs with extended resolution.

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