Summary
Multiplexed nucleic-acid detection is essential for molecular diagnostics and spatial genomics, but conventional fluorescence methods are often limited by spectral overlap, nonspecific signals, and restricted encoding capacity. We present a spatial fluorescence barcode (SFB) platform based on transiently luminescent DNA beads (TLDBs) that enables single-color, high-plex readout. In this method, targets are encoded through the spatial arrangement of DNA-functionalized beads, eliminating the need for multicolor labeling or spectral unmixing. Target recognition is achieved through toehold-mediated strand displacement, and built-in nucleases enable autonomous enzymatic resetting for repeated use of probes. The system employs monochromatic spatial encoding, decoupling encoding capacity from spe
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