Proper sample preparation, scan setup, data collection and image analysis are key factors in successful atomic force microscopy (AFM), which can avoid gloss phenomena effectively from unreasonable manipulations or instrumental defaults. Fresh cleaved mica and newly treated glass cover were checked first as the substrates for all of the sample preparation for AFM. Then, crystals contamination from buffer was studied separately or combined with several biologic samples, and the influence of scanner, scan mode and cantilever to data collection was also discussed intensively using molecular and cellular samples. At last, images treatment and analysis with off-line software had been focused on standard and biologic samples, and artificial glosses were highly considered for their high probability. SCANNING 31: 49-58, 2009. (c) 2009 Wiley Periodicals, Inc.
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