Transmission Mueller-matrix spectroscopic ellipsometry is applied to the cuticle of the beetle Cetonia aurata in the spectral range 300-1000 nm. The cuticle is optically reciprocal and exhibits circular Bragg filter features for green light. By using differential decomposition of the Mueller matrix, the circular and linear birefringence as well as dichroism of the beetle cuticle are quantified. A maximum value of structural optical activity of 560 degrees/mm is found. (C) 2016 Optical Society of America

A Mueller matrix of a sample can be used to determine the polarization of  reflected light  for  incident light with arbitrary polarization. The polarization can be quantified  in terms of ellipticity, polarization azimuth and degree of polarization. We apply spectroscopic Mueller-matrix ellipsometry at multiple angles of incidence  to study the cuticle of beetles and derive  polarization features for incident unpolarized light.  In particular we address chiral phenomena in scarab beetles,  the origin of their structural colors and the observed high degree of circular polarization is discussed. Results from beetles in the Scarabaeidae subfamilies Cetoniinae and Rutelinae are presented including specimens with broad-band silver- or gold-like colors with metallic shine as well as specimens with narrow-band green or red reflectors. The variation of polarization with angle of incidence and occurrence of both left-handed and right-handed polarization from a single species are presented. We also use Mueller-matrix spectra in electromagnetic modeling and show how to determine structural parameters including cuticle layer thicknesses and optical properties. Interference oscillations in the observed spectra are due to allowed optical modes and we show how to develop a structural model of a cuticle based on this effect. Sum decomposition of  Mueller matrices measured on a depolarizing cuticle of a beetle is briefly discussed.

Spectroscopic Mueller-matrix ellipsometry at variable angles of incidence is applied to beetle cuticles using a small (50 -100 μm) spot size. It is demonstrated how ellipticity and degree of polarization of the reflected light can be derived from a Mueller matrix providing a detailed insight into reflection properties. Results from Cetonia aurata, Chrysina argenteola and Cotinis mutabilis are presented. The use of Mueller matrices in regression analysis to extract structural and optical parameters of cuticles is briefly described and applied to cuticle data from Cetonia aurata whereby the pitch of the twisted layered structure in the cuticle is determined as well as the refractive indices of the epicuticle and the exocuticle.

The attractive shiny metallic colour of jewel scarabs is originating from the structure of the exoskeleton.For some directions and wavelengths of the incident light this structure will also cause the reflectedlight to have a large ellipticity (near-circular polarization). This is due to that the exoskeleton is ahelicoidal structure, formed by layers of chitin molecules. The reflected light is most commonly lefthandedpolarized but right-handed polarization is also observed. In this work six species of Scarabbeetles from the Chrysina genus are investigated. The complete Mueller-matrix is measured with adual rotating compensator ellipsometer (RC2, J.A.Woollam Co., Inc.). The results are presented ascontour plots where we represent different parameters as a function of incidence angle 2[25; 75]and wavelength 2[240; 1000]nm of the incident beam. Parameters of particular interest are the m41element of the Mueller-matrix, which is related to the circular polarization behaviour, the degree ofpolarization, the ellipticity and the absolute value of the azimuth angle. From ocular observationsthrough left- and right-circularly polarizing filters all specimens showed clear polarization effects interms of colour changes. However, the Mueller matrix ellipsometry measurements showed two generaltypes of polarization behaviour depending on the studied species. Chrysina macropus and Chrysinaperuviana had a smaller range of m41 values around zero. Much larger m41 variations were observedfor Chrysina argenteola, Chrysina chrysargyrea and Chrysina resplendens. Chrysina gloriosa hadboth types of polarization behaviour depending on if the measurements where made on the green orgolden parts of this striped beetle. Comparisons among samples of beetles from the same species wereconducted. For instance, different specimens of Chrysina resplendens show rather large differences inthe polarization response whereas specimens of Chrysina chrysargyrea showed very similar polarizationbehaviour. All studied specimens did in some sense reflect both right- and left-handed polarizedlight. In many cases very high ellipticities (near-circular polarization states) were observed. Modelsof structures generating the observed polarization effects as well as biological aspects will also bediscussed.Figure 257: Three pictures of C. chrysargyrea from left to right taken with aleft-circular polarizer, no filters and with a right-circular polarizer in front of thecamera. Two contour plots of m41 for C. chrysargyrea showing a large region withleft-handed near-circular polarization and C. resplendens showing a large regionwith right-handed near-circular polarization.

One hundred years ago Michelson discovered circular polarization in reflection from beetles. Today a novel Mueller-matrix ellipsometry setup allows unprecedented detailed characterization of the beetles polarization properties. A formalism based on elliptical polarization for description of reflection from scarab beetles is here proposed and examples are given on four beetles of different character: Coptomia laevis - a simple dielectric mirror; Cetonia aurata - a left-hand narrow- band elliptical polarizer; Anoplognathus aureus - a broad-band elliptical polarizer; and Chrysina argenteola - a left-hand polarizer for visible light at small angles, whereas for larger angles, red reflected light is right-handed polarized. We confirm the conclusion of previous studies which showed that a detailed quantification of ellipticity and degree of polarization of cuticle reflection can be performed instead of only determining whether reflections are circularly polarized or not. We additionally investigate reflection as a function of incidence angle. This provides much richer information for understanding the behaviour of beetles and for structural analysis.

Polarization effects in reflection from the cuticle of scarab beetles studied by spectroscopic Mueller-matrix ellipsometry

H. Arwin^*, T. Berlind, J. Birch, L. Fernandez Del Rio, J. Gustafson, J. Landin,

R. Magnusson, C. Åkerlind, and K. Järrendahl

Department of Physics, Chemistry and Biology, Linköping University, Sweden

^*corresponding author: han@ifm.liu.se

Abstract- Many scarab beetles exhibit structural colors and complex polarization phenomena in reflection. These effects are characterized with spectroscopic Mueller-matrix ellipsometry in our work. The polarization ellipse of reflected light as well as the degree of polarization is presented including variations with angle of incidence and wavelength. Emphasis is on beetles showing chiral effects and structural modeling of cuticle nanostructure is discussed.

Background Since one hundred years it is known that some scarab beetles reflect elliptically polarized light as demonstrated by Michelson for the beetle Chrysina resplendens [1]. The handedness of the polarization is in a majority of the cases left-handed but also right-handed polarization has been found [2,3]. The ellipticity varies with wavelength and viewing angle but can be close to +1 or -1 (right or left circular polarization, respectively) and in addition these beetles may exhibit beautiful structural colors. The polarization and color effects are generated in the outer part of the exoskeleton, the cuticle. These natural photonic structures are often multifunctional and play important roles for survival of beetles, e.g. for hiding from or scaring predators, for intraspecies communication, etc. [4]. However, such structures may find use in many commercial applications and a major motivation for detailed studies of natural photonic structures is that they inspire to biomimetic applications [5,6].

Approach Our objective is to use spectral Mueller-matrix data on scarab beetles to parameterize reflection properties in terms of polarization parameters and degree of polarization. The studied beetles all are phytophagous and include species from the Cetoniinae subfamily (e.g. Cetonia aurata and Coptomia laevis,), the Rutelinae subfamily (e.g. Chrysina argenteola and Chrysina resplendens) and the Melolonthinae subfamily (Cyphochilus insulanus). Furthermore, structural modeling is presented on Cetonia aurata and a few more beetles to demonstrate that structural parameters can be extracted by advanced modeling of Mueller-matrix data.

Experimental A dual rotating compensator ellipsometer (RC2, J. A. Woollam Co., Inc.) is used to record all 16 Mueller-matrix elements m_ij (i,j=1..4) in the spectral range 300 – 900 nm at angles of incidence in the range 20-70º. The elements are normalized to m₁₁ and thus have values between -1 and +1. All measurements are performed on the scutellum (a small triangular part on the dorsal side of the beetles) with focusing optics resulting in a spot size of the order of 50-100 mm. The software CompleteEASE (J. A. Woollam Co., Inc.) is used for analysis.

Results and discussion As an example, Fig. 1 shows contour plots of Mueller-matrix data measured on Cetonia aurata. This beetle has a metallic shine and if illuminated with unpolarized white light it reflects left-handed polarized green light as revealed by the non-zero Mueller-matrix elements m₁₄ and m₄₁ in the green spectral region for angles of incidence below about 45º. This is clearly seen in the graph to the right in Fig. 1 which shows a spectrum for Mueller-matrix element m₄₁at 20º as well as fitted model data. A model based on a twisted lamella structure, also called Bouligand structure, is used to model the chiral nanostructure [4]. Given the complexity of the nanostructure, an excellent model fit is achieved. The obtained model parameters are the spectral variation of the refractive index of the birefringent lamellas and the pitch. The model also includes a dielectric surface layer.

Fig.1. Left: Mueller-matrix data on Cetonia aurata. Each contour plot shows m_ij, where i and j correspond to the row and column, respectively. m₁₁ =1 and is not shown but is replaced with a photo of the beetle. Right: Experimental and model-generated Mueller-matrix element m₄₁at an angle of incidence of 20º.

From the Mueller-matrix data one can also determine so called derived parameters including azimuth and ellipticity of the polarization ellipse and the degree of polarization. The variations of these parameters with angle of incidence are presented for a selection of scarab beetles. Examples of both left-handed and right-handed polarization effects are shown and the importance of degree of polarization will be discussed.

Concluding remarks Mueller-matrix spectra at oblique incidence are very rich in information about reflection properties and allows parameterization of polarization parameters of the reflected light. Both left-handed and right-handed reflected light is found in scarab beetles. Mueller-matrix data can also be used for a detailed modeling of the nanostructure of the cuticle of beetles.

AcknowledgementsFinancial support was obtained from the Knut and Alice Wallenberg foundation and the Swedish Research Council. The Museum of Natural History in Stockholm, the National Museum of Natural Science in Madrid, the Berlin Museum of Natural History and the Natural History Museum in London are acknowledged for loan of beetles.

REFERENCES

1. Michelson, A. A. “On Metallic Colouring in Birds and Insects,” Phil. Mag., 21, 554-567, 1911.
2. Goldstein, D. H. “Polarization properties of Scarabaeidae,” Appl. Opt., 45, 7944-7950, 2006.
3. Hodgkinson, I., Lowrey, S., Bourke, L., Parker, A. and McCall, M. W. “Mueller-matrix characterization of beetle cuticle polarized and unpolarized reflections from representative architectures,” Appl. Opt., 49, 4558-4567, 2010.
4. Vukusic, P. and Sambles, J. R. “Photonic structures in biology,” Nature, 424, 852-855, 2003.
5. Lenau, T. and Barfoed, M. “Colours and Metallic Sheen in Beetle Shells - A Biomimetic Search for Material Structuring Principles Causing Light Interference,” Adv. Eng. Mat., 10, 299-314. 2008.
6. Parker, A. R. and Townley, H. E “Biomimetics of photonic nanostructures,” Nature Nanotech., 2, 347-351, 2007.

The Mueller matrix elements m_ij representing the polarization response from a nanostructured materialis determined by the constituent materials optical properties and the superstructure. Here, we investigate how chiral structures in form of helicoidally stacked uniaxial layers determine m_ij as a functionof polarization state, wavelength, incidence angle and azimuthal angle of the incoming light. The studied parameters include the layer materials ordinary/extraordinary optical properties, Euler angle values, and layer thickness as well as the thickness and pitch of the helicoidal superstructure. Sub- and superstructure inhomogeneity is also introduced. From the Fresnel-based calculations, m_ij aswell as the degree of polarization, ellipticity and azimuth of the polarization ellipse are obtained and presented as contour and trace plots to give a complete view of the polarization behavior. The results from the calculations are compared with Mueller matrix spectroscopic ellipsometry measurements of both natural and synthesized helicoidal structures. The measurements were performed with a dualrotating compensator system (RC2, J.A. Woollam Co., Inc.) for wavelengths in the range from 245 to 1000 nm and incident angles from 20 to 75°. For some measurements the azimuthal angle of the incident light was varied. The investigated natural chiral structures were exoskeletons from several beetles in the scarab subfamilies Cetoniinae and Rutelinae. As predicted from the calculations it isobserved that the reflection from these beetles can have a high degree of polarization and high ellipticity (near-circular polarization). Both left- and right-polarization was observed. The synthesized structures are helicoidal nanorods of Al_1−xIn_xN grown on sapphire substrates with metal-nitride seedlayers using UHV magnetron sputtering. Due to an internal composition gradient (a variation of x) in the crystalline structure, the nanorods will tilt away from the substrate normal. Helicoidal structures can thus be obtained by rotating the substrate around its normal during deposition. Samples with different pitch and layer thickness with right-handed as well as left-handed chirality were grown. Also for these structures both left and right near-circular polarized light is observed. By combining calculations, ellipsometry measurements and scanning electron microscopy characterization we get agood input to build layered models of the natural and synthetic samples. After regression fitting agood agreement between calculated and measured optical data were obtained.

Nature offers a plethora of possibilities for optical biomimetics. Some birds, butterflies, insects and other creatures exhibit brilliant colors as result of diverse optical phenomena produced by micro- and nanostructures at the near surface. Particularly, the shiny metallic colors reflected by the exoskeleton (the so called cuticle) of some beetles show elliptical polarization properties; most commonly of the left-handed type but also the right-handed type has been found [1]. These color and polarization properties have been related to a twisted plywood or Bouligand structure which is comprised by the clustering of chitin nanofibrils wrapped by proteins in a planar woven.

In this work we report the polarization properties of light reflected from the scarab beetle Cotinis mutabilis (Gory and Percheron 1833) and the microstructure of its cuticle. This species is found in Mexico and the southwestern part of the United States. The specimen under study was collected at the facilities of Cinvestav in Querétaro, Mexico where it is known as mayate (Mayatl in Náuhtl, the language of Aztecs). The Mayatl presents a green mate color in its dorsal side with red-orange stripes in the elytra. On the other hand, the abdominal side shows a shiny green metallic color which is subject of the present study. The polarization properties are investigated by the complete Mueller matrix (M) measured with a dual rotating compensator ellipsometer (J. A. Woollam Co., Inc.) at angles of incidence of 20-75° and wavelength range of 250-1000 nm. In particular, the elements m₄₁ and m₁₄ of M show that green left-handed polarized light is reflected when the beetle is illuminated at near-normal angles of incidence with unpolarized white light. As the angle of incidence increases the maximum of light reflected is blue-shifted. The degree of polarization, ellipticity, and azimuth calculated with relationships between m_j1 elements provide a full description of the polarization state of light reflected from the beetle’s cuticle for incident un-polarized light. Scanning electron microscopy images of the cuticle reveal that the epicuticle and exocuticle comprise a (hard) layer ca. 15 μm thick. Two regions can be differentiated in the exocuticle: the outer exocuticle without any clear structure and the inner exocuticle where a clear multilayer structure is observed. Beneath the inner exocuticle is the endocuticle which is comprised by unidirectional layers of microfibrils alternate with layers running at nearly right angles to each other in a pseudo-orthogonal arrangement. The cuticle also was imaged with a microscope coupled to a FTIR system allowing further identification of bands due to chitin.

In temporary wetlands in the River Dalalven floodplains, recurrent but irregular floods induce massive hatching of the flood-water mosquito Aedes sticticus, which causes enormous nuisance. Flood-water mosquito control using the biological larvicide Bacillus thuringiensis var. israelensis (Bti) was commenced in parts of the floodplains during 2002, and here we report the first six years of full-season monitoring of general insect emergence from temporary wetlands with and without treatment. Emergence traps, which were emptied weekly, were used from May to September each year. A total of 137,153 insects of 13 taxonomic orders were collected. Diptera was highly dominating and especially the sub-order Nematocera with 18 families was a very prominent taxon. Bti-treatment effects were analysed by taxonomic order, by sub-order in Diptera and Hemiptera, and by family for Nematocera and Coleoptera for the whole study period. We found no significant negative effects of Bti treatments on the production of insects by taxonomic order, with the exception of Coleoptera in the long term. However, no significant negative effects were found for the Coleoptera families, neither in the short term nor in the long term. There was no significant negative treatment effect on Nematocera production, neither when analyzed for the whole sub-order nor when analyzed by family. However, abundance of Ceratopogonidae was significantly higher in experimental than in reference wetlands. We conclude that Bti-treatment effects on insect production may be minute in comparison to other environmental factors structuring the insect fauna of the temporary wetlands studied.

Massive mosquito nuisance problems, caused by the flood-water mosquito Aedes sticticus, occur after floods in the flood plains of the River Dalälven, central Sweden. Since 2002, the biological mosquito larvicide Bacillus thuringiensis var. israelensis (Bti) has been used to control these mosquitoes. Here, we report results from the first six years of monitoring Chironomidae, the most susceptible non-target organisms, in three wetlands with Bti-treatment against mosquitoes and in three wetlands without treatment. Emergence traps were used for continuous insect sampling from May to September each year, 2002–2007, and were emptied once a week. A total of 21,394 chironomids of 135 species were collected, and the subfamily Orthocladiinae dominated the fauna followed by Chironominae and Tanypodinae. The annual chironomid production in the wetlands was low, with an average of 1917 individuals per m², and 42 g ash-free dry weight per m². We found no reduced production of chironomids at neither family, nor subfamily level, in Bti-treated as compared to untreated wetlands. This is the first long-term follow-up study of the possible effects of Bti-based mosquito larval control on chironomid species production. In the short-term view, one species had higher production in treated areas. In the long-term view, four species had higher and one species had lower production in treated areas. We conclude that the Bti-based control of flood-water mosquitoes does not cause any major direct negative effects on chironomid production, and therefore does not seem to induce any risk for indirect negative effects on birds, bats or any other predators feeding on chironomids.