Layout design is ubiquitous in daily life, an indispensable element behind successful visual guidance and aesthetic experience of 2-dimensional media or 3-dimensional spaces alike. Therefore, how different layouts and content lead to different user eye movement and emotional reactions becomes a vital subject for practical designs and aesthetic education, while relevant studies are few in number. The present research started off by discussing basic layout prototypes. Then an 8 × 2 factorial design experiment was employedbased on two factors, ＂layout design＂ and ＂content meaningfulness/meaninglessness.＂ It used eye tracking technique and Pleasure- Arousal-Dominance (PAD) emotion measurement scale to analyze browsing movements and aesthetic emotions of 40 participants. As the results showed, the two factors have notable interaction with Pleasure emotion; layout factor influences Arousal emotions, while content meaningfulness/meaninglessness influences Dominance emotion. Besides making theoretic contributions, the present research also hopes to provide clear guidance for design education and thus facilitate layout designs fitting the current multimedia platforms.