Food safety analysis plays a significant role in controlling food contamination and supervision. In recent years, multiplex optical bioassays (MOBAs) have been widely applied to analyze multiple hazards due to their efficiency and low cost. However, due to the challenges such as multiplexing capacity, poor sensitivity, and bulky instrumentation, the further application of traditional MOBAs in food screening has been limited. In this review, effective strategies regarding food safety MOBAs are summarized, such as spatial‐resolution modes performed in multi‐T lines/dots strips or arrays of strip/microplate/microfluidic chip/SPR chip and signal‐resolution modes employing distinguishable colorimetric/luminescence/fluorescence/surface plasma resonance/surface‐enhanced Raman spectrum as signal tags. Following this, new trends on how to design engineered sensor architecture and exploit distinguishable signal reporters, how to improve both multiplexing capacity and sensitivity, and how to integrate these formats into smartphones so as to be mobile are summarized systematically. Typically, in the case of enhancing multiplexing capacity and detection throughput, microfluidic array chips with multichannel architecture would be a favorable approach to overcome the spatial and physical limitations of immunochromatographic assay (ICA) test strips. Moreover, noble metal nanoparticles and single‐excitation, multiple‐emission luminescence nanomaterials hold great potential in developing ultrasensitive MOBAs. Finally, the exploitation of innovative multiplexing strategy hybridized with powerful and widely available smartphones opens new perspectives to MOBAs. In future, the MOBAs should be more sensitive, have higher multiplexing capacity, and easier instrumentation.
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