Nanoscale cuticle density variations correlate with pigmentation and color in butterfly wing scales

How pigment distribution correlates with cuticle density within a microscopic butterfly wing scale, and how both impact final reflected color remains unknown. We used ptychographic X-ray computed tomography to quantitatively determine, at nanoscale resolutions, the three-dimensional mass density of scales with pigmentation differences. By comparing cuticle densities with pigmentation and color within a scale, we determine that the lower lamina structure in all scales has the highest density and lowest pigmentation. Low pigment levels also correlate with sheet-like chitin structures as opposed to rod-like structures, and distinct density layers within the lower lamina help explain reflected color. We propose that pigments, in addition to absorbing specific wavelengths, can affect cuticle polymerization, density, and refractive index, thereby impacting reflected wavelengths that produce structural colors.