Optofluidic laser (OFL) technology, as an emerging technology combining microfluidics and laser technology, offers many unique advantages in sensing applications and has become a research hotspot for highly-sensitive intracavity biochemical analysis. Biochemical sensors based on OFLs can detect changes in biochemical parameters by using significant changes in laser output characteristics, so as to achieve high detection sensitivity. Here, we provide an overview of OFLs with a focus on their constructions, the design of OFL-based biochemical sensors, and their applications in biochemical analysis. Firstly, the three elements of an OFL, including the optical microcavity, gain medium, and pump source, are described systematically. After explaining the basic principles and characterization of OFLs for biochemical sensing, the current research progress of OFL-based biochemical sensors is summarized and analyzed according to the combination of OFLs with different assay techniques. This is followed by a discussion of the research on OFLs at the level of biological macromolecules, cells, and tissues. Finally, in view of the applications of OFLs in the field of biochemical sensing, the current challenges and future development directions are briefly discussed.