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Hyperspectral imaging—a technique that combines the high spectral resolution of spectrophotometry with the high spatial resolution of photography—holds great promise for the study of animal coloration. However, applications of hyperspectral imaging to questions about the ecology and evolution of animal color remain relatively rare. The approach can be expensive and unwieldy, and we lack user-friendly pipelines for capturing and analyzing hyperspectral data in the context of animal color. Fortunately, costs are decreasing and hyperspectral imagers are improving, particularly in their sensitivity to wavelengths (including ultraviolet) visible to diverse animal species. To highlight the potential of hyperspectral imaging for animal coloration studies, we developed a pipeline for capturing, sampling, and analyzing hyperspectral data (here, in the 325 nm to 700 nm range) using avian museum specimens. Specifically, we used the pipeline to characterize the plumage colors of the King bird-of-paradise (Cicinnurus regius), Magnificent bird-of-paradise (C.magnificus), and their putative hybrid, the King of Holland’s bird-of-paradise (C.magnificus x C.regius). We also combined hyperspectral data with 3D digital models to supplement hyperspectral images of each specimen with 3D shape information. Using visual system-independent methods, we found that many plumage patches on the hybrid King of Holland’s bird-of-paradise are—to varying degrees—intermediate relative to those of the parent species. This was true of both pigmentary and structurally colored plumage patches. Using visual system-dependent methods, we showed that only some of the differences in plumage patches among the hybrid and its parent species would be perceivable by birds. Hyperspectral imaging is poised to become the gold standard for many animal coloration applications: comprehensive reflectance data—across the entire surface of an animal specimen—can be obtained in a matter of minutes. Our pipeline provides a practical and flexible roadmap for incorporating hyperspectral imaging into future studies of animal color. 

Analyzing color and pattern in the context of motion is a central and ongoing challenge in the quantification of animal coloration. Many animal signals are spatially and temporally variable, but traditional methods fail to capture this dynamism because they use stationary animals in fixed positions. To investigate dynamic visual displays and to understand the evolutionary forces that shape dynamic colorful signals, we require cross-disciplinary methods that combine measurements of color, pattern, 3-dimensional (3D) shape, and motion. Here, we outline a workflow for producing digital 3D models with objective color information from museum specimens with diffuse colors. The workflow combines multispectral imaging with photogrammetry to produce digital 3D models that contain calibrated ultraviolet (UV) and human-visible (VIS) color information and incorporate pattern and 3D shape. These “3D multispectral models” can subsequently be animated to incorporate both signaler and receiver movement and analyzed in silico using a variety of receiver-specific visual models. This approach—which can be flexibly integrated with other tools and methods—represents a key first step toward analyzing visual signals in motion. We describe several timely applications of this workflow and next steps for multispectral 3D photogrammetry and animation techniques.