Doekele G. Stavenga

The wing–scale morphologies of the pierid butterflies Pieris rapae (small white) and Delias nigrina (common jezabel), and the heliconine Heliconius melpomene are compared and related to the wing–reflectance spectra. Light scattering at the wing scales determines the wing reflectance, but when the scales contain an absorbing pigment, reflectance is suppressed in the absorption wavelength range of the pigment. The reflectance of the white wing areas of P. rapae, where the scales are studded with beads, is considerably higher than that of the white wing areas of H. melpomene, which has scales lacking beads. The beads presumably cause the distinct matt–white colour of the wings of pierids and function to increase the reflectance amplitude. This will improve the visual discrimination between conspecific males and females.

SUMMARY The effect of the UV-absorbing sensitizing pigment of fly photoreceptors on absolute, spectral and angular sensitivity was investigated with a wave-optics model for the facet lens-rhabdomere system. When sky light was used as a UV-rich light source, one sensitizing pigment molecule per rhodopsin increased the photoreceptor absorption by 14-18% with respect to pure rhodopsin, whilst two sensitizing pigment molecules per rhodopsin increased the absorption by 20-27%. Upon light adaptation, when the pupil mechanism is activated, photoreceptor absorption decreases; in the housefly, Musca, by up to 6-fold. The fully light-adapted pupil diminishes the photoreceptor9s acceptance angle by a factor of ∼0.6 due to selective absorption of higher order waveguide modes. Spatial acuity of dark-adapted photoreceptors is more or less constant throughout the visual wavelength range, including the UV, because the waveguide optics of the rhabdomere compromise acuity least at wavelengths most limited by diffraction of the facet lens. Diffraction is not the general limiting factor causative for UV sensitivity of insect eyes. Visual acuity is governed by diffraction only with a fully light-adapted pupil, which absorbs higher waveguide modes. Closure of the blue-absorbing pupil causes a UV-peaking spectral sensitivity of fly photoreceptors. The sensitizing pigment does not play an appreciable role in modifying spatial acuity, neither in the dark- nor the light-adapted state, due to the dominant contribution of green light in natural light sources.

A wave optics model for the facet lens-rhabdomere system of fly eyes is used to analyze the dependence of the angular and spectral sensitivity of R1–6 photoreceptors on the pupil mechanism. This assembly of light-absorbing pigment granules in the soma interacts with the waveguide modes propagating in the rhabdomere. A fly rhabdomere carries two modes in the middle wavelength range and four modes at short wavelengths, depending on the rhabdomere diameter and the angle of the incident light flux. The extension of the mode to outside the rhabdomere strongly depends on wavelength, and this dependence plays a determinant role in the light control function of the pupil. The absorbance spectrum of the pigment in the pupil granules is severely depressed at short wavelengths by waveguide effects, resulting in a distinct blue peak. Accordingly, pupil closure suppresses the photoreceptor’s spectral sensitivity much more in the blue-green than in the UV. The pupil only narrows the angular sensitivity at short wavelengths. The geometrical size of the rhabdomere governs the angular sensitivity of fly photoreceptors in the dark-adapted state, but diffraction takes over in the fully light-adapted state.

A wave-optical model for the integrated facet lens-rhabdomere system of fly eyes is used to calculate the effective light power in the rhabdomeres when the eye is illuminated with a point light source or with an extended source. Two rhabdomere types are considered: the slender rhabdomeres of R7,8 photoreceptors and the wider, but tapering R1–6 rhabdomeres. The angular sensitivities of the two rhabdomere types have been calculated as a function of F-number and wavelength by fitting Gaussian functions to the effective light power. For a given F-number, the angular sensitivity broadens with wavelength for the slender rhabdomeres, but it stays approximately constant for the wider rhabdomeres. The integrated effective light power increases with the rhabdomere diameter, but it is for both rhabdomere types nearly independent of the light wavelength and F-number. The results are used to interpret the small F-number of Drosophila facet lenses. Presumably the small head puts a limit to the size of the facet lens and favors a short focal length.

Three optical components of a fly's eye determine the angular sensitivity of the photoreceptors: the light diffracting facet lens, the wave-guiding rhabdomere and the light-absorbing visual pigment in the rhabdomere. How the integrated optical system of the fly eye shapes the angular sensitivity curves is quantitatively analyzed in five steps: (1) scalar diffraction theory for low Fresnel-number lenses is applied to four different facet lenses, with diameter 10, 20, 40, and 80 µm, respectively, assuming a constant F-number of 2.2; (2) optical waveguide theory is used to calculate waveguide modes propagating in circular cylindrical rhabdomeres with diameter 1.0, 2.0, and 4.0 µm, respectively; (3) the excitation of waveguide modes is studied with the tip of the waveguide positioned in the focal plane as well as outside this plane; (4) the light absorption from the various propagated modes by the visual pigment in the rhabdomere is calculated as a function of the angle of the incident light wave; and (5) the angular sensitivity of the photoreceptor is obtained by normalizing the total light absorption. Four wavelengths are considered: 300, 400, 500 and 600 nm. The analysis shows that the wavelength dependency of the lens diffraction is strongly compensated by that of the waveguide modes, an effect which is further enhanced by the decrease in light absorption when the mode number increases. The angular sensitivity of fly photoreceptors is robust to defocus and largely wavelength independent for all except very slender rhabdomeres.

Many insect species have darkly coloured eyes, but distinct colours or patterns are frequently featured. A number of exemplary cases of flies and butterflies are discussed to illustrate our present knowledge of the physical basis of eye colours, their functional background, and the implications for insect colour vision. The screening pigments in the pigment cells commonly determine the eye colour. The red screening pigments of fly eyes and the dorsal eye regions of dragonflies allow stray light to photochemically restore photoconverted visual pigments. A similar role is played by yellow pigment granules inside the photoreceptor cells which function as a light-controlling pupil. Most insect eyes contain black screening pigments which prevent stray light to produce background noise in the photoreceptors. The eyes of tabanid flies are marked by strong metallic colours, due to multilayers in the corneal facet lenses. The corneal multilayers in the gold-green eyes of the deer fly Chrysops relictus reduce the lens transmission in the orange-green, thus narrowing the sensitivity spectrum of photoreceptors having a green absorbing rhodopsin. The tapetum in the eyes of butterflies probably enhances the spectral sensitivity of proximal long-wavelength photoreceptors. Pigment granules lining the rhabdom fine-tune the sensitivity spectra.

SUMMARY: The eye shine of butterflies from a large number of ommatidia was observed with a modified epi-illumination apparatus equipped with an objective lens of large numerical aperture. A few representative cases are presented: the satyrine Bicyclus anynana, the heliconian Heliconius melpomene, the small white Pieris rapae and the small copper Lycaena phlaeas. The colour of the eye shine is determined mainly by the reflectance spectrum of the tapetal mirror and the transmittance spectrum of the photoreceptor screening pigments, if present near the light-guiding rhabdom. Reflectance spectra measured from individual ommatidia show that tapetum and screening pigments are co-expressed in fixed combinations, thus determining different ommatidial classes. The classes are distributed in an irregular pattern that can be rapidly assessed with the novel epi-illumination apparatus. Many butterfly species appear to have red-reflecting ommatidia,which is interpreted to indicate the presence of red-sensitive photoreceptors.

The regional characteristics of the eyes of butterflies from different families have been surveyed using epi-illumination microscopy, utilizing the eyeshine visible due to the tapetum situated proximally to the rhabdom. All butterflies studied have a high spatial acuity in the frontal region. The facet diameter varies slightly across the eye, and the interommatidial angle and the eye parameter p are especially large dorsally. Whereas the ommatidial lattice is generally highly regular, the eyeshine colours distinctly depend on the species. Sometimes the eyeshine is locally uniform, but often it is heterogeneous. It is hypothesized that the regional characteristics as well as the local heterogeneity are adaptations that optimize spectral discrimination.