We present a preliminary investigation of macroscopic polarimetric imaging of uterine cervix. Orthogonal state contrast (OSC) images of healthy and anomalous cervices have been taken in vivo at 550 nm. Four ex vivo cervix samples have been studied in full Muller polarimetry, at 550 nm and 700 nm, and characterized in detail by standard pathology. One sample was totally healthy, another one carried CIN lesions at very early stage (CIN1) in its visible exocervical region, while for the other two samples more advanced (CIN3) lesions were present, together with visible glandular epithelium (ectropion). Significant birefringence has been observed in the healthy regions of all six samples, both in vivo and ex vivo. Standard treatments of the Mueller images of the ex vivo samples allowed to quantify both retardation and depolarization. Retardation reached 60° in healthy regions, and disappeared in the anomalous regions of the other three ex vivo samples. The depolarization power was largest in healthy regions, and lower in CINs and ectropion. Possible origins of the observed effects are briefly discussed.
Combining optical microscopy, synchrotron X-ray diffraction and ellipsometry, we studied the internal structure of linear defect domains (oily streaks) in films of a smectic liquid crystal 8CB with thicknesses in the range of 100-300 nm. These films are confined between air and a rubbed PVA polymer substrate which imposes hybrid anchoring conditions (normal and unidirectional planar, respectively). We show how the presence or absence of dislocations controls the structure of highly deformed thin smectic films. Each domain contains smectic layers curved in the shape of flattened hemicylinders to satisfy both anchoring conditions, together with grain boundaries whose size and shape are controlled by the presence of dislocation lines. A flat grain boundary normal to the interface connects neighboring hemicylinders, while a rotating grain boundary (RGB) is located near the axis of curvature of the cylinders. The RGB shape appears such that dislocation lines are concentrated at its summit close to the air interface. The smectic layers reach the polymer substrate via a transition region where the smectic layer orientation satisfies the planar anchoring conditions over the entire polymer substrate and whose thickness does not depend on that of the film. The strength of planar anchoring appears to be high, larger than 10(-2) mJ m(-2), compensating for the high energy cost of creating an additional 2D defect between a horizontal smectic layer and perpendicular ones of the transition region. This 2D defect may be melted, in order to avoid the creation of a transition region structure composed of a large number of dislocations. As a result, linear defect domains can be considered as arrays of oriented defects, straight dislocations of various Burger vectors, whose location is now known, and 2D nematic defects. The possibility of easy variation between the present structure with a moderate amount of dislocations and a structure with a large number of dislocations is also demonstrated.
We studied the azimuthal orientations of collagen fibers in histological slides of uterine cervical tissue by two different microscopy techniques, namely Mueller polarimetry (MP) and Second Harmonic Generation (SHG). SHG provides direct visualization of the fibers with high specificity, which orientations is then obtained by suitable image processing. MP provides images of retardation (among other polarimetric parameters) due to the optical anisotropy of the fibers, which is enhanced by Picrosirius Red staining. The fiber orientations are then assumed to be those of the retardation slow axes. The two methods, though fully different from each other, provide quite similar maps of average fiber orientations. Overall, our results confirm that MP microscopy provides reliable images of dominant fiber orientations at a much lower cost that SHG, which remains the "gold standard" for specific imaging of collagen fibers using optical microscopy.
Reflected Mueller matrix spectra were measured and simulated for asymmetrical photoresist master diffraction gratings in conical mounting (i.e., the direction of grating grooves was not perpendicular to the plane of light incidence). From the electromagnetic reciprocity theorem, Mueller matrix of symmetric grating (composed of only reciprocal materials, and operating in zeroth-order diffraction) is invariant under transposition (M ¼ M t). For zeroth-order diffraction of asymmetric gratings, the lack of profile rotational symmetry violates this reciprocity and, consequently, breaks the symmetry of the above-mentioned matrix. This property of the Mueller matrix of asymmetric gratings was experimentally observed and numerically modeled at all experimental illumination conditions with the exception of planar mounting (the direction of grating grooves was perpendicular to the plane of light incidence), where there is no cross-polarization effect for the gratings composed of isotropic materials. It was demonstrated that optical nonreciprocity of diffraction gratings can be used for unambiguous detection of grating profile asymmetry. In addition, choosing optimal measurement configuration (i.e., azimuthal angles) considerably increases the sensitivity of the detection technique. V
International audienceAngle resolved Mueller polarimetry implemented as polarimetric imaging of a back focal plane of a high NA microscope objective has already demonstrated a good potential for CD metrology. Here we present the experimental and numerical results indicating that this technique may also be competitive for the measurements of overlay error delta. A series of samples of superimposed gratings with well controlled overlay errors have been manufactured and measured with the angle resolved Mueller polarimeter. The overlay targets were 20-mu m wide. When the overlay error is delta is equal to 0, absolute values of elements of real 4x4 Mueller matrix M are invariant by matrix transposition. Otherwise this symmetry breaks down. Consequently, we define the following overlay estimator matrix as E = |M| - |M|(t). The simulations show that matrix element E-14 is the most sensitive to the overlay error. The scalar estimator of E-14 was calculated by averaging the pixel values over a specifically chosen mask. This estimator is found to vary linearly with d for overlay values up to 50 nm. Our technique allows entering small overlay marks (down to 5-mu m wide). Only one target measurement is needed for each overlay direction. The actual overlay value can be determined without detailed simulation of the structure provided two calibrated overlay structures are available for each direction
We present a polarimetric instrument suitable for the simultaneous measurement of angle resolved normalized Mueller matrices for polar angles ranging from 0 degrees to 60 degrees and all azimuths. The polarimetric modulation and analysis are performed by means of an optimized polarization state generator and analyzer based on nematic liquid crystals. A high numerical aperture (0.95) microscope objective is used in double pass to illuminate the sample, with its rear focal plane imaged on a low noise CCD. This polarimeter can be used either in reflection, with the sample set in the objective front focal plane, or in transmission, for thin transparent samples. This latter configuration, which involves an additional spherical mirror with its center of curvature at the objective front focus, is described in detail. This instrument was used to accurately determine the directions of the optic axes and the angular dependence of the retardation of a biaxial polyethylene terephthalate (PET) plastic substrate in spite of the strong depolarization essentially due to the source 10 nm spectral width or the limitation in angular resolution due to the pixels distribution of the CCD combined with the sample large retardation.
Polarized microscopy provides unique information on anisotropic samples. In its most complete implementation, namely Mueller microscopy, this technique is well suited for the visualization of fibrillar proteins orientations, with collagen in the first place. However, the intrinsic optical anisotropy of unstained tissues has to be enhanced by Picrosirius Red (PR) staining to enable Mueller measurements. In this work, we compared the orientation mapping provided by Mueller and second harmonic generation (SHG) microscopies on PR stained samples of vaginal and uterine cervix tissues. SHG is a multiphoton technique that is highly specific to fibrillar collagen, and was taken as the "gold standard" for its visualization. We showed that Mueller microscopy can be safely used to determine collagen orientation in PR stained cervical tissue. In contrast, in vaginal samples, Mueller microscopy revealed orientations not only of collagen but also of other anisotropic structures. Thus PR is not fully specific to collagen, which necessitates comparison to SHG microscopy in every type of tissue. In addition to this study of PR specificity, we determined the optimal values of the staining parameters. We found that staining times of 5 min, and sample thicknesses of 5 µm were sufficient in cervical and vaginal tissues.
We designed and built Matrix Distributed ECR (MDECR) PECVD reactor dedicated for dielectric filters deposition and equipped it with multiple sensors for process control. Planar matrix geometry of plasma source is based on electron cyclotron resonance effect at 2.45 GHz microwave frequency and provides scalability of the deposition on large area substrates. High (up to 5 nm/sec) deposition rate obtained due to high dissociation efficiency and careful design of the gas injection system. Optical emission spectroscopy, quadrupole mass-spectrometry and spectroscopic and multichannel kinetic ellipsometry are installed for in-situ studies and control of the film deposition. We performed studies of the nature of high-density plasma discharge in silane, oxygen and nitrogen mixture and correlated its properties with optical and physical properties of deposited materials. To demonstrate the capabilities, a wide band gradient index antireflection coating on glass was realized by deposition of SiO x N y alloy thin films. The predefined variation of an index in a profile is obtained by changing the flows of precursors. Real-time control is performed with multi-channel kinetic ellipsometry.
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