Current strategies for follow up of small animal (such as murine) models of liver disease are flawed to continuously monitor disease progression in the tissue level, and necessitate sacrifice of animals for tissue sampling. In this study, we aimed at developing a safe repetitive tool for sampling liver in steatosis and hepatitis C virus (HCV) models in tree shrews, by utilization of a minimally invasive surgery method. The biopsy of one animal was divided into two parts; they were immediately placed in formaldehyde and liquid nitrogen respectively. Repeated biopsy was performed with the same animal every one month. For subsequent biopsy, the liver tissue was collected in the different site. Hematoxylin and eosin (HE) stained for all histological evaluation. Total cellular RNA from liver tissue was extracted and reversely transcribed into cDNA, using the SuperScriptâ…¢ Reverse Transcription System (Invitrogen). This technique allows safe and repeated liver biopsies in tree shrews, yielding adequate tissue for histological staining and RNA extraction. We suggest that application of this approach may provide the investigator with the tools to assess and manipulate disease progression in vivo in individual tree shrew, allowing for a more comprehensive understanding of mechanisms of disease progression.