Biomedical image analysis has become critically important to the public health and welfare. However, analyzing biomedical images is time-consuming and labor-intensive, and has long been performed manually by highly trained human experts. As a result, there has been an increasing interest in applying machine learning to automate biomedical image analysis. Recent progress in deep learning research has catalyzed the development of machine learning in learning discriminative features from data with minimum human intervention. Many deep learning models have been designed and achieved superior performance in various data analysis applications. This chapter starts with the basic of deep learning models and some practical strategies for handling biomedical image applications with limited data. After that, case studies of deep feature extraction for gene expression pattern image annotations, imaging data completion for brain disease diagnosis, and segmentation of infant brain tissue images are discussed to demonstrate the effectiveness of deep learning in biomedical image analysis.