Photonic crystal (PC) based bioassay techniques have many advantages in sensitive biomolecular screening, label-free detection, real-time monitoring of enzyme activity, cell morphology research, and so on. This study provides an overall survey of the basic concepts and up-to-date research concerning the very promising use of PC materials for bioassays. It includes the design and application of PC films, PC microcarriers, PC fibers, and PC optofluidics for fluorescence enhancement or label-free bioassays. Emphasis is given to the description of the functional structures of different PC materials and their respective sensing mechanisms. Examples of detecting various types of analytes are presented. This article promotes communication among chemistry, biology, medicine, pharmacy, and material science.