The rapid development of healthcare techniques encourages the emergence of new molecular imaging agents and modalities. Fluorescence imaging that enables precise monitoring and detection of biological processes/diseases is extensively investigated as this imaging technique has strengths in terms of high sensitivity, excellent temporal resolution, low cost, and good safety. Aggregation-induced emission luminogens (AIEgens) have recently emerged as a new class of emitters that possess several notable features, such as high brightness, large Stokes shift, marked photostability, good biocompatibility, and so on. So far, AIEgens are widely explored and exhibit superb performance in the area of biomedicine and life sciences. Herein, this review summarizes and discusses the recent investigations of AIEgens for in vivo diagnosis and therapy including long-term tracking, 3D angiography, multimodality imaging, disease theranostics, and activatable sensing. Collectively, these results reveal that AIEgens are of great promise for in vivo biomedical applications. It is hoped that this review will lead to new insights into the development of advanced healthcare materials.