Multimodal semantic comprehension has attracted increasing research interests recently such as visual question answering and caption generation. However, due to the data limitation, fine-grained semantic comprehension has not been well investigated, which requires to capture semantic details of multimodal contents. In this work, we introduce “YouMakeup”, a large-scale multimodal instructional video dataset to support fine-grained semantic comprehension research in specific domain. YouMakeup contains 2,800 videos from YouTube, spanning more than 420 hours in total. Each video is annotated with a sequence of natural language descriptions for instructional steps, grounded in temporal video range and spatial facial areas. The annotated steps in a video involve subtle difference in actions, products and regions, which requires fine-grained understanding and reasoning both temporally and spatially. In order to evaluate models’ ability for fined-grained comprehension, we further propose two groups of tasks including generation tasks and visual question answering from different aspects. We also establish a baseline of step caption generation for future comparison. The dataset will be publicly available at https://github.com/AIM3-RUC/YouMakeup to support research investigation in fine-grained semantic comprehension.