The spot auto-focusing (AF) method with a unique high-definition auto-correlation function (HD-ACF) proposed in the previous paper is improved and is now applicable to general specimens at a wide range of magnifications. According to the definition where the AF is defocused to obtain the highest resolution, the proposed method achieves the sharpest HD-ACF profile in the AF spot image. The relationship where the sharpest HD-ACF profile gives the highest resolution is theoretically explained, and practical AF examples for different specimens and magnifications are experimentally demonstrated. Specimens include a yeast cell thin section at 10-k magnification, a standard grating replica used as a ruler at 50-k, a crystal lattice of graphitized carbon at 400-k and a 60°-tilted thin section (yeast cell) at 10-k. Different procedures are prepared to actively identify the defocus position that gives the sharpest HD-ACF profile. Every AF result demonstrates the highest-resolution image.
Keywords: Fourier ring correlation; auto-correlation function; auto-correlation radial distribution function; electron tomography; high-definition auto-correlation function; spot auto-focusing.
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