Coloration principles of nymphaline butterflies - thin films, melanin, ommochromes and wing scale stacking.

The coloration of the common butterflies Aglais urticae (small tortoiseshell), Aglais io (peacock) and Vanessa atalanta (red admiral), belonging to the butterfly subfamily Nymphalinae, is due to the species-specific patterning of differently coloured scales on their wings. We investigated the scales' structural and pigmentary properties by applying scanning electron microscopy, (micro)spectrophotometry and imaging scatterometry. The anatomy of the wing scales appears to be basically identical, with an approximately flat lower lamina connected by trabeculae to a highly structured upper lamina, which consists of an array of longitudinal, parallel ridges and transversal crossribs. Isolated scales observed at the abwing (upper) side are blue, yellow, orange, red, brown or black, depending on their pigmentation. The yellow, orange and red scales contain various amounts of 3-OH-kynurenine and ommochrome pigment, black scales contain a high density of melanin, and blue scales have a minor amount of melanin pigment. Observing the scales from their adwing (lower) side always revealed a structural colour, which is blue in the case of blue, red and black scales, but orange for orange scales. The structural colours are created by the lower lamina, which acts as an optical thin film. Its reflectance spectrum, crucially determined by the lamina thickness, appears to be well tuned to the scales' pigmentary spectrum. The colours observed locally on the wing are also due to the degree of scale stacking. Thin films, tuned pigments and combinations of stacked scales together determine the wing coloration of nymphaline butterflies.

Stavenga DG ，Leertouwer HL ，Wilts BD 《-》

Longwing (Heliconius) butterflies combine a restricted set of pigmentary and structural coloration mechanisms.

Longwing butterflies, Heliconius sp., also called heliconians, are striking examples of diversity and mimicry in butterflies. Heliconians feature strongly colored patterns on their wings, arising from wing scales colored by pigments and/or nanostructures, which serve as an aposematic signal. Here, we investigate the coloration mechanisms among several species of Heliconius by applying scanning electron microscopy, (micro)spectrophotometry, and imaging scatterometry. We identify seven kinds of colored scales within Heliconius whose coloration is derived from pigments, nanostructures or both. In yellow-, orange- and red-colored wing patches, both cover and ground scales contain wavelength-selective absorbing pigments, 3-OH-kynurenine, xanthommatin and/or dihydroxanthommatin. In blue wing patches, the cover scales are blue either due to interference of light in the thin-film lower lamina (e.g., H. doris) or in the multilayered lamellae in the scale ridges (so-called ridge reflectors, e.g., H. sara and H. erato); the underlying ground scales are black. In the white wing patches, both cover and ground scales are blue due to their thin-film lower lamina, but because they are stacked upon each other and at the wing substrate, a faint bluish to white color results. Lastly, green wing patches (H. doris) have cover scales with blue-reflecting thin films and short-wavelength absorbing 3-OH-kynurenine, together causing a green color. The pigmentary and structural traits are discussed in relation to their phylogenetic distribution and the evolution of vision in this highly interesting clade of butterflies.

Wilts BD ，Vey AJM ，Briscoe AD ，Stavenga DG ... -  《BMC EVOLUTIONARY BIOLOGY》

Brilliant iridescence of Morpho butterfly wing scales is due to both a thin film lower lamina and a multilayered upper lamina.

Butterflies belonging to the nymphalid subfamily, Morphinae, are famous for their brilliant blue wing coloration and iridescence. These striking optical phenomena are commonly explained as to originate from multilayer reflections by the ridges of the wing scales. Because the lower lamina of the scales of related nymphalid butterflies, the Nymphalinae, plays a dominant role in the wing coloration, by acting as a thin film reflector, we investigated single blue scales of three characteristic Morpho species: M. epistrophus, M. helenor and M. cypris. The experimental data obtained by spectrophotometry, scatterometry and scanning electron microscopy demonstrated that also in the Morpho genus the lower lamina of both the cover and ground scales acts as an optical thin film reflector, contributing importantly to the blue structural coloration of the wings. Melanin pigment has a contrast-enhancing function in a sub-class of ground scales.

Giraldo MA ，Stavenga DG 《-》

Glass scales on the wing of the swordtail butterfly Graphium sarpedon act as thin film polarizing reflectors.

The wings of the swordtail butterfly Graphium sarpedon (the Common Bluebottle) have blue/green-colored patches that are covered on the underside by two types of scales: white and glass scales. Transmission and scanning electron microscopy revealed that the white scales are classically structured: the upper lamina, with prominent ridges and large open windows, is well separated by trabeculae from a flat, continuous lower lamina. In the glass scales, the upper lamina, with inconspicuous ridges and windows, is almost flat and closely apposed to the equally flat lower lamina. The glass scales thus approximate ideal thin films, in agreement with the observation that they reflect light directionally and are iridescent. Reflectance and transmittance spectra measured from the glass scales with a microspectrophotometer agree with spectra calculated for an ideal non-absorbing thin film. Imaging scatterometry of single, isolated glass scales demonstrated that the reflected light can be strongly polarized, indicating that they function as polarizing reflectors.

Stavenga DG ，Matsushita A ，Arikawa K ，Leertouwer HL ，Wilts BD ... -  《-》

Wing coloration and pigment gradients in scales of pierid butterflies.

Depending on the species, the individual scales of butterfly wings have a longitudinal gradient in structure and reflectance properties, as shown by scanning electron microscopy and microspectrophotometry. White scales of the male Small White, Pieris rapae crucivora, show a strong gradient in both the density in pigment granules and the reflectance. After pigment extraction by aqueous ammonia, scales of male P. r. crucivora closely resemble the unpigmented scales of female P. r. crucivora. Only a minor gradient exists in the white and orange scales of the male Orange Tip, Anthocharis cardamines. Pigment extraction of orange scales of A. cardamines causes bleaching. Partial bleaching transforms the scales so that they resemble certain scales of Phoebis philea that have a natural extreme gradient. Reflectance measurements on an artificial stack of two overlapping scales as well as on the scale stacks existing on intact and partially denuded wings of the Large White, Pieris brassicae, quantitatively demonstrate the reflectance enhancement by scale stacking.

Giraldo MA ，Stavenga DG 《-》