Background: Post-occupancy evaluation (POE) can play a critical role in evidence-based healthcare design (EBD) by evaluating design effectiveness and adding to the knowledge base. However, its potential has not been fully realized due to the lack of standardized tools, inaccessibility of findings, a focus on technical performance instead of patient-centred outcomes, and irrelevance to healthcare processes. Objective: To strengthen the EBD knowledge base by developing standardized POE tools. Methodology: A conceptual framework was created from a review of over 100 research publications. A standardized POE toolkit was developed, including questionnaires on design strategies, healthcare outcomes, and staff perception. Pilot testing was conducted at five healthcare construction projects. This toolkit is being further augmented by additional on-site audit POE tools. Findings: From the pilot testing of the first POE tool, significant variations were found among the five pilot facilities in terms of the design strategies adopted. Satisfaction with the indoor environment was higher among occupants at facilities with more sustainable design strategies implemented. Patients reported high satisfaction (65–83% top ratings) and high willingness to recommend facilities (71–85% ‘definitely recommend’). The general satisfaction of staff with building ranged from 0.88 to 2.2 on a scale from −3 to 3. Conclusion: A robust framework and a set of standard tools for obtaining broad building-level data related to healthcare design have been established to comprehensively evaluate building performance and to enable quick dissemination and easy access to new knowledge created.