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I -IK staining influenced specimen shrinkage, micro-CT image quality, and pathological image quality in a concentration- and time-dependent manner. After comprehensively considering these indicators, 3% I -IK staining for 48 and 12h were found to be optimal for rabbit tongue tissues and TSCC samples, respectively. This method could provide a detailed 3-D structure of TSCC samples compared with HE sections. Moreover, tumor and normal tissues could be differentiated by their glycogen content, which has high affinity with I -IK. I -IK