Andrej Meglič

Extreme polarization sensitivity in the retina of the corn borer moth Ostrinia

ABSTRACT The visual system of the European corn borer (Ostrinia nubilalis) was analysed with microscopy and electrophysiological methods (electroretinograms and single-cell recordings). Ostrinia nubilalis has a pair of mainly ultraviolet-sensitive ocelli and a pair of compound eyes, maximally sensitive to green light. The ommatidia contain a tiered, fused rhabdom, consisting of the rhabdomeres of 9–12 photoreceptor cells with sensitivity peak wavelengths at 356, 413, 480 and 530 nm. The photoreceptors in a large dorsal rim area have straight rhabdomeres and high polarisation sensitivity (PS1,2=3.4, 14). Elsewhere, in the main retina, the majority of photoreceptors have non-aligned microvilli and negligible PS, but each ommatidium contains one or two blue-sensitive distal photoreceptors with straight microvilli parallel to the dorsoventral axis, yielding extremely high PS (PS1,2,3=56, 63, 316). Rhabdoms containing distal cells with potentially high PS have evolved at least twice: in moths (Crambidae, Noctuidae, Saturniidae), as well as in dung beetles (Scarabaeidae). The distal photoreceptors with high PS, sensitive to vertically polarised light, represent a monopolatic system, which is unsuitable for the proper analysis of electric field vector (e-vector) orientation. However, the distal photoreceptors might be used in conjunction with polarisation-insensitive photoreceptors to detect objects that reflect polarised light with stereotyped orientation. Summary: Polarisation vision in the moths is not confined to the dorsal rim area; the retina of Crambid, Noctuid and Saturniid moths and some dung beetles contains distal photoreceptors with extremely high polarisation sensitivity.

A fast multispectral light synthesiser based on LEDs and a diffraction grating

Optical experiments often require fast-switching light sources with adjustable bandwidths and intensities. We constructed a wavelength combiner based on a reflective planar diffraction grating and light emitting diodes with emission peaks from 350 to 630 nm that were positioned at the angles corresponding to the first diffraction order of the reversed beam. The combined output beam was launched into a fibre. The spacing between 22 equally wide spectral bands was about 15 nm. The time resolution of the pulse-width modulation drivers was 1 ms. The source was validated with a fast intracellular measurement of the spectral sensitivity of blowfly photoreceptors. In hyperspectral imaging of Xenopus skin circulation, the wavelength resolution was adequate to resolve haemoglobin absorption spectra. The device contains no moving parts, has low stray light and is intrinsically capable of multi-band output. Possible applications include visual physiology, biomedical optics, microscopy and spectroscopy.

Electrochemical inactivation of cyanobacteria and microcystin degradation using a boron-doped diamond anode — A potential tool for cyanobacterial bloom control

Cyanobacterial blooms are global phenomena that can occur in calm and nutrient-rich (eutrophic) fresh and marine waters. Human exposure to cyanobacteria and their biologically active products is possible during water sports and various water activities, or by ingestion of contaminated water. Although the vast majority of harmful cyanobacterial products are confined to the interior of the cells, these are eventually released into the surrounding water following natural or artificially induced cell death. Electrochemical oxidation has been used here to damage cyanobacteria to halt their proliferation, and for microcystin degradation under in-vitro conditions. Partially spent Jaworski growth medium with no addition of supporting electrolytes was used. Electrochemical treatment resulted in the cyanobacterial loss of cell-buoyancy regulation, cell proliferation arrest, and eventual cell death. Microcystin degradation was studied separately in two basic modes of treatment: batch-wise flow, and constant flow, for electrolytic-cell exposure. Batch-wise exposure simulates treatment under environmental conditions, while constant flow is more appropriate for the study of boron-doped diamond electrode efficacy under laboratory conditions. The effectiveness of microcystin degradation was established using high-performance liquid chromatography-photodiode array detector analysis, while the biological activities of the products were estimated using a colorimetric protein phosphatase-1 inhibition assay. The results indicate potential for the application of electro-oxidation methods for the control of bloom events by taking advantage of specific intrinsic ecological characteristics of bloom-forming cyanobacteria. The applicability of the use of boron-doped diamond electrodes in remediation of water exposed to cyanobacteria bloom events is discussed.

Classical lepidopteran wing scale colouration in the giant butterfly-moth Paysandisia archon

The palm borer moth Paysandisia archon (Castniidae; giant butterfly-moths) has brown dorsal forewings and strikingly orange-coloured dorsal hindwings with white spots surrounded by black margins. Here, we have studied the structure and pigments of the wing scales in the various coloured wing areas, applying light and electron microscopy and (micro)spectrophotometry, and we analysed the spatial reflection properties with imaging scatterometry. The scales in the white spots are unpigmented, those in the black and brown wing areas contain various amounts of melanin, and the orange wing scales contain a blue-absorbing ommochrome pigment. In all scale types, the upper lamina acts as a diffuser and the lower lamina as a thin film interference reflector, with thickness of about 200 nm. Scale stacking plays an important role in creating the strong visual signals: the colour of the white eyespots is created by stacks of unpigmented blue scales, while the orange wing colour is strongly intensified by stacking the orange scales.

Development and plasticity of mitochondria and electrical properties of the cell membrane in blowfly photoreceptors

Blowfly photoreceptors are highly energy demanding sensory systems. Their information processing efficiency is enabled by the high temporal resolution of the cell membrane, requiring heavy metabolic support by the mitochondria. We studied the developmental changes of the mitochondrial apparatus and electrical properties of the photoreceptor membrane in the white eyed Calliphora vicina Chalky. Using in vivo microspectrophotometry and Western blot analysis, we found an age-dependent increase in the concentration of mitochondrial pigments. The maximal change occurred during the first week. The age-related changes were smaller in dark-bred than in light-bred flies. The mitochondrial pigment content increased after the switch from dark to light rearing and decreased after the switch from light to dark rearing. The electrical parameters of the photoreceptors were investigated with intracellular recordings. The resting membrane resistance and time constant decreased significantly after eclosion. The decrease was again most significant during the first week of adult life, paralleled with changes in the Na/K pump-dependent hyperpolarizing afterpotential. We conclude that the photoreceptor mitochondria exhibit remarkable ontogenetic and phenotypic plasticity, because the quantity of mitochondrial pigments tightly follows the development of the cell membrane as well as the energy demands of the photoreceptors under different rearing conditions.

The effects of electrochemical oxidation on in-vivo fluorescence and toxin content in Microcystis aeruginosa culture

The increasing occurrence of cyanobacterial blooms in water bodies is a serious threat to the environment. Efficient in-lake treatment methods for the control of cyanobacteria proliferation are needed, their in-vivo detection to obtain a real-time response to their presence, as well as the information about their physiological state after the applied treatment. In-vivo fluorescence measurements of photosynthetic pigments have proved to be effective for quantitative and qualitative detection of phytoplankton in a water environment. In the experiment, chlorophyll and phycocyanin fluorescence sensors were used concurrently to detect stress caused by electrochemical oxidation applying an electrolytic cell equipped with borondoped diamond electrodes on a laboratory culture of cyanobacteria Microcystis aeruginosa PCC 7806. The inflicted injuries were reflected in a clear transient increase in the phycocyanin fluorescence signal (for 104 %± 43%) 24 h after the treatment, which was not the case for the chlorophyll fluorescence signal. In the next 72 h of observation, the fluorescence signals decreased (on 40% of the starting signal) indicating a reduction of cell number, which was confirmed by cell count (24% reduction of the starting concentration) and analysis of extracted chlorophyll and phycocyanin pigment. These results demonstrate the viability of the combined application of two sensors as a useful tool for in-vivo detection of induced stress, providing real-time information needed for the evaluation of the efficiency of the in-lake treatment and decision upon the necessity of its repetition. The electrochemical treatment also resulted in a lower free microcystins concentration compared to control.

The Fly Sensitizing Pigment Enhances UV Spectral Sensitivity While Preventing Polarization-Induced Artifacts

Microvillar photoreceptors are intrinsically capable of detecting the orientation of e-vector of linearly polarized light. They provide most invertebrates with an additional sensory channel to detect important features of their visual environment. However, polarization sensitivity (PS) of photoreceptors may lead to the detection of polarization-induced false colors and intensity contrasts. Most insect photoreceptors are thus adapted to have minimal PS. Flies have twisted rhabdomeres with microvilli rotated along the length of the ommatidia to reduce PS. The additional UV-absorbing sensitizing pigment on their opsin minimizes PS in the ultraviolet. We recorded voltage from Drosophila photoreceptors R1–6 to measure the spectral dependence of PS and found that PS in the UV is invariably negligible but can be substantial above 400 nm. Using modeling, we demonstrate that in R1–6 without the sensitizing pigment, PS in the UV (PSUV) would exceed PS in the visible part of the spectrum (PSVIS) by a factor PSUV/PSVIS = 1.2–1.8, as lower absorption of Rh1 rhodopsin reduces self-screening. We use polarimetric imaging of objects relevant to fly polarization vision to show that their degree of polarization outdoors is highest in the short-wavelength part of the spectrum. Thus, under natural illumination, the sensitizing pigment in R1–6 renders even those cells with high PS in the visible part unsuitable for proper polarization vision. We assume that fly ventral polarization vision can be mediated by R7 alone, with R1–6 serving as an unpolarized reference channel.