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The identification and differentiation of a large number of distinct molecular species with high temporal and spatial resolution is a major challenge in biomedical science. Fluorescence microscopy is a powerful tool, but its multiplexing ability is limited by the number of spectrally distinguishable fluorophores. Yin and coworkers demonstrated the construction of submicrometer nanorods that can be used as fluorescent barcodes [84]. Barcodes can be modified easily to display sequence‐specific attachment sites through simple DNA‐origami staple extensions. Spatial control over the positioning of fluorophores was achieved on the surface of a DNA nanorod (Figure 1.10e). The fluorescent dye‐conjugated DNA strands (imager strands) transiently bind their specific target sites on the DNA nanorod, which produces “blinking” for super‐resolution reconstruction, similar to a previous report [82]. Using epifluorescence or TIRF microscopy, 216 distinct barcodes were unambiguous decoded. On the other hand, barcodes with higher spatial information density were demonstrated via the construction of super‐resolution barcodes (Figure 1.10f,g). One type of barcode was used to tag yeast surface receptors, which suggests their potential applications as in situ imaging probes for diverse biomolecular and cellular entities in their native environments.