The Sir Colin MacKenzie Sanctuary is of state significance due to its historical associations with Sir Colin MacKenzie, and the development of medical research in the state. MacKenzie's cottage is the last building in the sanctuary constructed by Sir Colin, and is therefore of great significance as a direct link to MacKenzie's work at the site. The sanctuary is also of state significance as a place of education, and as one of Victoria's most prominent tourist destinations.

The Mackenzie Cottage, Healesville Sanctuary is historically and scientifically important for its close associations with Sir Colin MacKenzie and the sanctuary he fostered. It is important as the last remaining tangible link to the earliest era of the development of the research field station in 1920. MacKenzie is a critically significant figure in the history of the understanding and protection of Australian native wildlife. As the earliest building at the Healesville Sanctuary it shares in the enormous social significance of the sanctuary to Victorians and visitors to the State as a place of education, recreation and fascination. (Victorian Heritage Register)

Physical Conditions: Excellent

Integrity: Evidence of stages