Comic page layouts directly affects the communication of the story. It can be described as the blueprint for the comic page design, and it is an important step in the comic creation process. This study is an empirical research using eye tracking technology to measure subjects. It explores the influence of the number of panels, the shape of panels, and the widths of gutters of manga page layouts on readability. The research shows that the number of panels does affect the readability of manga page layouts. The pages with fewer panels are less readable. The reason is speculated that the layout with less divisions provide less information to readers. The impact of the panel-shapes and the gutter-widths on readability of manga page layouts needs further research and verification. In addition, the eye tracking experiments also found that many readers do not follow the reading path set by the creator. However, whether they follow the default path does not affect the readers’ understanding of the story. Readers’ reading sequence for comics is also affected by their personal reading experience. Readers with more experience in reading American comics, Korean comics, and webtoons, as well as readers who are accustomed to reading online comics, comic e-books with electronic products, have a greater chance of reading comics in a way going against the “reverse Z-path” principle; Readers who have been reading manga magazines and manga tankobon (books published for independent volumes) for a long time usually follow the “reverse Z-path” reading principle of manga.