Abstract: A fluorescent identification and focusing target indicia and associated methods for use. The indicia is used to label a substrate, and may subsequently be used first as a target to focus the optical analysis and next as a specific identifier of the substrate and/or of reagents used with the substrate. The label may be a separate layer joined to the substrate by an adhesive. The indicia may include multiple dyes, a dye that produces fluorescence in a plurality of channels, a reflective component, a human interpretable component, a quantification means, a sizing standard, or other components.