Figure 3. Dual-labeled-zone fluorescent barcodes.

(a) Typical TIRF microscopy images of five selected barcode species, shown both in separate channels and as superimposed images. Scale bar: 5 μm. (b) Statistics obtained by analyzing two 50 × 50 μm² images of each barcode species (~85 barcodes, the exact sample size N is noted beside the corresponding bar-graph). The barcode types are noted under the horizontal axis of the diagram. Color scheme (unrelated to the pseudo-colors of the fluorophores): blue, correct barcodes (correct geometry and color identity); green, incorrect barcodes (correct geometry but incorrect color identity); red, monomer nanorods (one spot or two touching spots); purple, barcodes with wrong geometry (bending angle <120°, see Methods in SI). (c) Computer-aided barcode counting results of the 72-barcode pool (N=2,617) and the 216-barcode pool (N=7,243), where the number of qualified barcodes are plotted as bar-graphs (see Supplementary Tables S6 and S7 for numbers). A computer-generated reference barcode image is placed underneath the corresponding bar. Note that the same horizontal axis is used for both bar-graph diagrams. (d) A table containing one representative TIRF image (1.4 × 0.7 μm²) for each of the 216 dual-labeled-zone barcode species.