Polarization neutral point pairs of the solar corona and the lunar disc observed during the total solar eclipse on 11 August 1999 in Hungary.

During the total solar eclipses on 11 August 1999 in Kecel (Hungary) and on 29 March 2006 in Side (Turkey), two Hungarian groups performed full-sky imaging polarimetric measurements of the eclipsed sky. They observed the spatiotemporal change of the celestial polarization pattern and detected three polarization neutral points as well as two points with maximal polarization of the sky. Parallel to these studies, the polarization pattern in front of the lunar disc, the solar corona, and the surrounding sky have also been measured. During the total solar eclipse on 21 August 2017 in the USA (Rexburg-Idaho, Madras-Oregon), three American/international groups have measured the polarization characteristics of the full sky and the solar corona. The first group observed the spatiotemporal variation of the celestial polarization pattern, and the second group detected three polarization neutral points of the sky and observed two neutral points of the solar corona. The latter were named as Minnaert and van de Hulst neutral points. The third group observed two neutral points of the lunar disc. We have reanalyzed the earlier polarization patterns of the lunar disc, solar corona, and the surrounding sky measured during the Hungarian total eclipse on 11 August 1999. In these reanalyzed polarization patterns, all four neutral points observed during the eclipse on 21 August 2017 in the USA occur: the Minnaert/van de Hulst neutral point pair above/below the eclipsed Sun, where coronal polarization is canceled by sky polarization, and the northern and southern (unnamed) neutral points of the lunar disc, where the directions of polarization of coronal light and foreground skylight are perpendicular to each other with the same polarized intensity. We name the latter two polarization neutral points after Coulson and Vorobiev.

On the trail of Vikings with polarized skylight: experimental study of the atmospheric optical prerequisites allowing polarimetric navigation by Viking seafarers.

Between AD 900 and AD 1200 Vikings, being able to navigate skillfully across the open sea, were the dominant seafarers of the North Atlantic. When the Sun was shining, geographical north could be determined with a special sundial. However, how the Vikings could have navigated in cloudy or foggy situations, when the Sun's disc was unusable, is still not fully known. A hypothesis was formulated in 1967, which suggested that under foggy or cloudy conditions, Vikings might have been able to determine the azimuth direction of the Sun with the help of skylight polarization, just like some insects. This hypothesis has been widely accepted and is regularly cited by researchers, even though an experimental basis, so far, has not been forthcoming. According to this theory, the Vikings could have determined the direction of the skylight polarization with the help of an enigmatic birefringent crystal, functioning as a linearly polarizing filter. Such a crystal is referred to as 'sunstone' in one of the Viking's sagas, but its exact nature is unknown. Although accepted by many, the hypothesis of polarimetric navigation by Vikings also has numerous sceptics. In this paper, we summarize the results of our own celestial polarization measurements and psychophysical laboratory experiments, in which we studied the atmospheric optical prerequisites of possible sky-polarimetric navigation in Tunisia, Finland, Hungary and the high Arctic.

Confocal fluorescence detected linear dichroism imaging of isolated human amyloid fibrils. Role of supercoiling.

Amyloids are highly organized insoluble protein aggregates that are associated with a large variety of degenerative diseases. In this work, we investigated the anisotropic architecture of isolated human amyloid samples stained with Congo Red. This was performed by fluorescence detected linear dichroism (FDLD) imaging in a laser scanning confocal microscope that was equipped with a differential polarization attachment using high frequency modulation of the polarization state of the laser beam and a demodulation circuit. Two- and three-dimensional FDLD images of amyloids provided information on the orientation of the electric transition dipoles of the intercalated Congo Red molecules with unprecedented precision and spatial resolution. We show that, in accordance with linear dichroism imaging (Jin et al. Proc Natl Acad Sci USA 100:15294, 2003), amyloids exhibit strong anisotropy with preferential orientation of the dye molecules along the fibrils; estimations on the orientation angle, of around 45°, are given using a model calculation which takes into account the helical organization of the filaments and fibrils. Our data also show that FDLD images display large inhomogeneities, high local values with alternating signs and, in some regions, well identifiable µm-sized periodicities. These features of the anisotropic architecture are accounted for by supercoiling of helically organized amyloid fibrils.

Imaging anisotropy using differential polarization laser scanning confocal microscopy.

We have constructed differential polarization (DP) attachments to a laser scanning microscope (LSM) for imaging the main DP quantities of anisotropic microscopic objects. The DP-LSM operates with high-frequency modulation and subsequent demodulation and displays the main DP quantities pixel by pixel. These, for linearly polarized light, include: (i) linear birefringence (LB), which is exhibited by structurally and/or optically anisotropic material; (ii) linear dichroism (LD), which carries information on the anisotropic distribution of the molecules, i.e. of their absorbance transition dipole vectors, in the sample; (iii) fluorescence-detected LD (FDLD), which carries the same information for fluorescent dyes upon excitations with two orthogonally polarized light beams; (iv) anisotropy of the fluorescence emission (r), excited with non-polarized light, which is determined by the distribution of the emission transition dipole vectors in the sample and is analogous with LD and (v) the degree of polarization of the fluorescence emission (P), excited with polarized light, which depends on the depolarization of the emission e.g. due to the rotation of molecules during their excitation lifetimes. In fluorescence regimes, the DP images can be recorded in the confocal regime of the microscope, which thus warrants good spatial resolution and the possibility of mapping the anisotropy in three dimensions. In this paper, we outline the design and technical realization of our DP-LSM and give a few examples on DP imaging of different biological samples.

Some new faces of membrane microdomains: a complex confocal fluorescence, differential polarization, and FCS imaging study on live immune cells.

Lipid rafts are cholesterol- and glycosphingolipid-rich plasma membrane microdomains, which control signal transduction, cellular contacts, pathogen recognition, and internalization processes. Their stability/lifetime, heterogeneity remained still controversial, mostly due to the high diversity of raft markers and cellular models. The correspondence of the rafts of living cells to liquid ordered (Lo) domains of model membranes and the effect of modulating rafts on the structural dynamics of their bulk membrane environment are also yet unresolved questions. Spatial overlap of various lipid and protein raft markers on live cells was studied by confocal laser scanning microscopy, while fluorescence polarization of DiIC18(3) and Bodipy-phosphatidylcholine was imaged with differential polarization CLSM (DP-CLSM). Mobility of the diI probe under different conditions was assessed by fluorescence correlation spectroscopic (FCS). GM1 gangliosides highly colocalized with GPI-linked protein markers of rafts and a new anti-cholesterol antibody (AC8) in various immune cells. On the same cells, albeit not fully excluded from rafts, diI colocalized much less with raft markers of both lipid and protein nature, suggesting the Lo membrane regions are not equivalents to lipid rafts. The DP-CLSM technique was capable of imaging probe orientation and heterogeneity of polarization in the plasma membrane of live cells, reflecting differences in lipid order/packing. This property--in accordance with diI mobility assessed by FCS--was sensitive to modulation of rafts either through their lipids or proteins. Our complex imaging analysis demonstrated that two lipid probes--G(M1) and a new anti-cholesterol antibody--equivocally label the membrane rafts on a variety of cell types, while some raft-associated proteins (MHC-II, CD48, CD59, or CD90) do not colocalize with each other. This indicates the compositional heterogeneity of rafts. Usefulness of the DP-CLSM technique in imaging immune cell surface, in terms of lipid order/packing heterogeneities, was also shown together with its sensitivity to monitor biological modulation of lipid rafts.

Imaging fluorescence detected linear dichroism of plant cell walls in laser scanning confocal microscope.

Anisotropy carries important information on the molecular organization of biological samples. Its determination requires a combination of microscopy and polarization spectroscopy tools. The authors constructed differential polarization (DP) attachments to a laser scanning microscope in order to determine physical quantities related to the anisotropic distribution of molecules in microscopic samples; here the authors focus on fluorescence-detected linear dichroism (FDLD). By modulating the linear polarization of the laser beam between two orthogonally polarized states and by using a demodulation circuit, the authors determine the associated transmitted and fluorescence intensity-difference signals, which serve the basis for LD (linear dichroism) and FDLD, respectively. The authors demonstrate on sections of Convallaria majalis root tissue stained with Acridin Orange that while (nonconfocal) LD images remain smeared and weak, FDLD images recorded in confocal mode reveal strong anisotropy of the cell wall. FDLD imaging is suitable for mapping the anisotropic distribution of transition dipoles in 3 dimensions. A mathematical model is proposed to account for the fiber-laminate ultrastructure of the cell wall and for the intercalation of the dye molecules in complex, highly anisotropic architecture.

Domains of Importin-alpha2 required for ring canal assembly during Drosophila oogenesis.

Null-mutation in Drosophila importin-alpha2, such as the deficiency imp-alpha2(D14), causes recessive female sterility with the formation of dumpless eggs. In imp-alpha2(D14) the transfer of nurse cell components to the oocyte is interrupted and the Kelch protein, an oligomeric ring canal actin organizer, is normally produced but fails to associate with the ring canals resulting in their occlusion. To define domains regulating Kelch deposition on ring canals we performed site-directed mutagenesis on protein binding domains and putative phosphorylation sites of Imp-alpha2. Phenotypic analysis of the mutant transgenes in imp-alpha2(D14) revealed that mutations affecting the Imp-beta binding-domain, the dimerization domain, and specific serine residues of putative phosphorylation sites led to a normal or nearly normal oogenesis but arrested early embryonic development, whereas mutations in the nuclear localization signal (NLS) and CAS/exportin binding domains resulted in ring canal occlusion and a drastic nuclear accumulation of the mutant proteins. Deletion of the Imp-beta binding domain also gave rise to a nuclear localization of the mutant protein, which partially retained its function in ring canal assembly. Thus, we propose that mutations in NLS and CAS binding domains affect the deposition of Kelch onto the ring canals and prevent the association of Imp-alpha2 with a negative regulator of Kelch function.

Alignment of biological microparticles by a polarized laser beam.

The optical alignment of biological samples is of great relevance to microspectrometry and to the micromanipulation of single particles. Recently, Bayoudh et al. (J. Mod. Opt. 50:1581-1590, 2003) have shown that isolated, disk-shaped chloroplasts can be aligned in a controlled manner using an in-plane-polarized Gaussian beam trap, and suggested that this is due to their nonspherical shape. Here we demonstrate that the orientation of various micrometer-sized isolated biological particles, trapped by optical tweezers, can be altered in a controlled way by changing the plane of linear polarization of the tweezers. In addition to chloroplasts, we show that subchloroplast particles of small size and irregular overall shape, aggregated photosynthetic light-harvesting protein complexes as well as chromosomes can be oriented with the linearly polarized beam of the tweezers. By using a laser scanning confocal microscope equipped with a differential polarization attachment, we also measured the birefringence of magnetically oriented granal chloroplasts, and found that they exhibit strong birefringence with large local variations, which appears to originate from stacked membranes. The size and sign of the birefringence are such that the resulting anisotropic interaction with the linearly polarized laser beam significantly contributes to the torque orienting the chloroplasts.

Neutral points of skylight polarization observed during the total eclipse on 11 August 1999.

We report here on the observation of unpolarized (neutral) points in the sky during the total solar eclipse on 11 August 1999. Near the zenith a neutral point was observed at 450 nm at two different points of time during totality. Around this celestial point the distribution of the angle of polarization was heterogeneous: The electric field vectors on the one side were approximately perpendicular to those on the other side. At another moment of totality, near the zenith a local minimum of the degree of linear polarization occurred at 550 nm. Near the antisolar meridian, at a low elevation another two neutral points occurred at 450 nm at a certain moment during totality. Approximately at the position of these neutral points, at another moment of totality a local minimum of the degree of polarization occurred at 550 nm, whereas at 450 nm a neutral point was observed, around which the angle-of-polarization pattern was homogeneous: The electric field vectors were approximately horizontal on both sides of the neutral point.

How Celestial Polarization Changes due to Reflection from the Deflector Panels Used in Deflector Loft and Mirror Experiments Studying Avian Orientation.

In bird experiments which investigate the role of optical orientation cues associated with sunset it is a common method to reflect the celestial sunset factors by vertical deflector panels. Similar panels are also used in the deflector loft of homing pigeons that rotate wind and light cues in a clockwise or counterclockwise direction. The responses of test birds in these behavioural experiments often appear to be contradictory, because the deflectors produce a polarized optical stimulus that differs both qualitatively and quantitatively from the natural skylight polarization. This paper gives a quantitative account of the change of celestial polarization in such situations. The polarization pattern of skylight reflected from different vertical deflector panels is computed as a function of the elevation of the sun from the horizon. It is shown that the reflection polarization of a deflector panel composed of transparent glass with or without white or black background cannot simulate the distribution of polarization of light in clear sky. On the other hand, if the deflector consists of a metal plate or metal-glass mirror, it can mimic the celestial polarization pattern well, because it changes the polarization of incident light after reflection only slightly.