As society ages, the need for online resources for older adults is becoming increasingly significant. However, because older adults are characterized by declining cognitive responses and delayed operating behaviors, they face many challenges in their access experience on healthcare service websites. Taking several domestic healthcare service websites as samples, this study analyzes the inclusiveness of website pages and detects accessibility from the perspective of inclusiveness by comparing the standards of Web Content Accessibility Guidelines (WCAG) and using an open source automated detection tool. The study aims to reveal the barriers faced by elderly users in accessing healthcare service websites from multiple dimensions such as perceivability, operability, comprehensibility and stability. At the same time, the web operating behavior, subjective feelings, and actual access barriers of elderly users were explored in depth through observation and in-depth interview methods. Based on the obtained data, accessibility optimization strategies for medical service websites are proposed to enable older users to access medical information and services more easily and conveniently.