Backgroundand goal: Micro-computed tomography (micro-CT) is an emerging technology in the biomedical field and enables us to analyze 3D structures non-destructively and observe these structures in various directions, thus enabling innovation in this area of pathology. However, application of micro-CT for medicine has just started and optimization per purpose has not yet been done. The purpose of this study is to 1) demonstrate the potential utility of micro-CT in pathology; 2) optimize micro-CT imaging technology and develop a standard operating protocol and; 3) investigate whether micro-CT incurs any radiation damage to pathological tissue samples.Material and methods:The samples of fresh tissue, formalin fixed tissue and formalin fixed paraffin-embedded (FFPE) tissue blocks were scanned using a custom-built Nikon Metrology micro-CT system with a variety of parameters then evaluated with histology correlation in detail. Radiation damage to tissue samples was also evaluated. Through our study, we have established the scanning protocol and workflow for each type of sample. Results:For fresh/fixed tissue, the house made polystyrene foam container was most ideal and the scanning time for fresh tissue was six minutes at as shortest, in which it is possible to detect neoplastic lesions in the tissue. In case of FFPE blocks, 10 -17 hours scanned images