Quantitative Birefringence Measurements Research Articles

Unsupervised topological analysis of polarized light microscopy: application to quantitative birefringence imaging.

The determination of birefringence (magnitude and axis orientation) of optical materials is of significant interest in various fields. In the case of composite samples, this task becomes complicated and time-consuming; therefore, a partially automated procedure for reconstructing birefringence spatial distribution becomes valuable. Herein, we propose a procedure to reconstruct the spatial distributions of the retardance and optical axis orientation in a geological thin section from sparse quantitative birefringence measurements, using automatic boundary detection on cross-polarized light microscopy images. We examine two particular areas on the selected geological thin section: one that presents a uniaxial crystal with a circular cross-section of its refractive index ellipsoid and the other with grains of varying orientations. The measurement gives the orientation of the grain's optical axis both in and out of the plane of the thin section, which explains the qualitative observations with the cross-polarized light microscope. Future work will connect the measured orientation of the rock thin section with its 3D geological orientation in the field.

Mitotic Index Determination on Live Cells From Label-Free Acquired Quantitative Phase Images Using a Supervised Autoencoder.

This interdisciplinary work focuses on the interest of a new auto-encoder for supervised classification of live cell populations growing in a thermostated imaging station and acquired by a Quantitative Phase Imaging (QPI) camera. This type of camera produces interferograms that have to be processed to extract features derived from quantitative linear retardance and birefringence measurements. QPI is performed on living populations without any manipulation or treatment of the cells. We use the efficient new autoencoder classification method instead of the classical Douglas-Rachford method. Using this new supervised autoencoder, we show that the accuracy of the classification of the cells present in the mitotic phase of the cell cycle is very high using QPI features. This is a very important finding since we demonstrate that it is now possible to very precisely follow cell growth in a non-invasive manner, without any bias. No dye or any kind of markers are necessary for this live monitoring. Any studies requiring analysis of cell growth or cellular response to any treatment could benefit from this new approach by simply monitoring the proportion of cells entering mitosis in the studied cell population.

Femtosecond laser direct writing in SiO2‐Al2O3 binary glasses and thermal stability of Type II permanent modifications

AbstractWe investigate the potential of fabricating thermally stable refractive index contrasts using femtosecond (fs) near‐infrared (IR) radiation in aluminosilicate glasses. A set of pure SiO2‐Al2O3 glasses are manufactured, characterized (density and Raman), and investigated after being irradiated by fs laser within the Type II regime. The formation of nanogratings is identified and studied using quantitative birefringence measurements. Their thermal stability is then investigated through 30 minutes step isochronal annealing (up to 1250°C). For both SiO2 and 50SiO2‐50Al2O3 compositions, the normalized birefringence does not decrease when tested up to 1100°C, while for the 4,6 mol% GeO2‐SiO2 erased for 20% at 1000°C.

Applications of shear-induced polarized light imaging (SIPLI) technique for mechano-optical rheology of polymers and soft matter materials

A new experimental method for studying the mechano-optical rheology of polymeric liquids and soft matter materials is presented. The method is based on a combination of rotational rheology and a recently developed optical technique—shear-induced polarized light imaging (SIPLI). The method provides a unique opportunity to monitor a complete sample view during rheological measurements in plate–plate and cone-and-plate geometry. Applications of the method are presented including simultaneous SIPLI and the rheology of the oriented lamellar phase of block copolymers and liquid crystals as well as a study of the thermally induced reversible transformation of worm-like micelles to spherical micelles. In addition, a direct relation between the shish formation and the polymer melt viscosity upturn during flow-induced crystallization of semi-crystalline polymers is demonstrated. An application of SIPLI for quantitative birefringence measurements is also shown. © 2016 The Authors. Journal of Polymer Science Part B: Polymer Physics Published by Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016, 54, 2151–2170

Nanogratings formation in multicomponent silicate glasses

We demonstrate the formation of porous nanogratings in various oxide glasses including TiO2-doped silica, GeO2 and alumino-borosilicate by near-IR femtosecond laser radiation. ULE and GeO2 glasses exhibit similar birefringence to pure silica, whereas Borofloat 33 reveals twice weaker amplitude. Using quantitative birefringence measurements, small-angle X-ray scattering and scanning electron microscopy, we correlate birefringence and porous nanolayers formation according to laser repetition rate and glass composition. We show that heat accumulation is a crucial parameter limiting the glass decomposition and thus nanogratings formation.

Quantitative retardance imaging of biological samples using quadriwave lateral shearing interferometry.

We describe a new technique based on the use of a high-resolution quadri-wave lateral shearing interferometer to perform quantitative linear retardance and birefringence measurements on biological samples. The system combines quantitative phase images with varying polarization excitation to create retardance images. This technique is compatible with living samples and gives information about the local retardance and structure of their anisotropic components. We applied our approach to collagen fibers leading to a birefringence value of (3.4 ± 0.3) · 10(-3) and to living cells, showing that cytoskeleton can be imaged label-free.

Structure of blown films of polyethylene/polybutene‐1 blends

AbstractThe structure of blown films of blends of low‐density polyethylene (PE‐LD) and isotactic polybutene‐1 (iPB‐1) with different content of iPB‐1 was investigated using wide‐ and small‐angle X‐ray scattering (WAXS and SAXS), transmission electron microscopy (TEM), and polarizing optical microscopy (POM). TEM proves formation of a matrix–particle phase structure due to immiscibility of the blend components. Within the iPB‐1 particles, needle‐like crystals with c‐axis orientation were observed. The PE‐LD matrix showed two populations of crystals. WAXS data indicate that the majority of crystals were oriented with the c‐axis perpendicular to machine direction (MD), while SAXS data prove additional presence of stacks of lamellae, oriented parallel to MD. Quantitative birefringence measurements showed that the majority of molecule segments were oriented in the direction of the circumference of the film, confirming the WAXS data. The crystal orientation has direct impact on mechanical properties, which was demonstrated by measurement of the anisotropy of the modulus of elasticity. POLYM. ENG. SCI., 2009. © 2009 Society of Plastics Engineers

Analysis of enamel microbiopsies in shed primary teeth by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM)

The aims of this study were 1) to verify how close to the theoretically presumed areas are the areas of enamel microbiopsies carried out in vivo or in exfoliated teeth; 2) to test whether the etching solution penetrates beyond the tape borders; 3) to test whether the etching solution demineralizes the enamel in depth. 24 shed upper primary central incisors were randomly divided into two groups: the Rehydrated Teeth Group and the Dry Teeth Group. An enamel microbiopsy was performed, and the enamel microbiopsies were then analyzed by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM). Quantitative birefringence measurements were performed. The “true” etched area was determined by measuring the etched enamel using the NIH Image analysis program. Enamel birefringence was compared using the paired t test. There was a statistically significant difference when the etched areas in the Rehydrated teeth were compared with those of the Dry teeth ( p = 0.04). The etched areas varied from − 11.6% to 73.5% of the presumed area in the Rehydrated teeth, and from 6.6% to 61.3% in the Dry teeth. The mean percentage of variation in each group could be used as a correction factor for the etched area. Analysis of PM pictures shows no evidence of in-depth enamel demineralization by the etching solution. No statistically significant differences in enamel birefringence were observed between values underneath and outside the microbiopsy area in the same tooth, showing that no mineral loss occurred below the enamel superficial layer. Our data showed no evidence of in-depth enamel demineralization by the etching solution used in the enamel microbiopsy proposed for primary enamel. This study also showed a variation in the measured diameter of the enamel microbiopsy in nineteen teeth out of twenty four, indicating that in most cases the etching solution penetrated beyond the tape borders.

Rheo-optical measurements of the first and third normal stresses of homopolymer poly(vinyl methyl ether) melt

Normal stresses play a key role in polymer processing, yet accurate measurements are still challenging. Simultaneous rheo-optical measurements are conducted on a poly(vinyl methyl ether) homopolymer melt over a wide range of temperatures and oscillatory shear frequencies, in an effort to measure the normal stresses, by using quantitative flow birefringence measurements. The stress optical rule holds well for this polymer as expected, with the value of the stress optic coefficient of (6.38±0.19)×10−11 cm2/dyn at 30°C. The first and third normal stress difference coefficients\(\psi ^{ * }_{1} ,\psi ^{ * }_{3} \), calculated using a single memory constitutive equation applied to the stress and birefringence data, are in excellent agreement. The ratio of the measured third and first normal stress difference coefficients, (1−β)=0.71±0.05, agrees well with the result of the Doi–Edwards model with independent alignment approximation (β=0.28). The measurement of normal stress difference coefficients with such small deviations proves the robust nature of the improved rheo-optical instrument and its ability to measure complete stress tensor.

Monitoring rejection after heart transplantation: cytoimmunological monitoring on blood cells and quantitative birefringence measurements on endomyocardial biopsy specimens.

Cytoimmunological monitoring and quantitative birefringence measurements were used as potential aids in diagnosing acute rejection after heart transplantation instead of histopathological assessment of the endomyocardial biopsy specimen alone. Cytoimmunological monitoring was based on morphological inspection and quantitation of mononuclear cells, particularly activated lymphoid cells. Quantitative birefringence measurements comprise a variable for myocyte contractile function. Its read out is the ratio of the degree of birefringence before contraction to that after. Cytoimmunological monitoring indicated significantly higher concentrations of activated lymphocytes in moderate or severe acute rejection, and quantitative birefringence measurements indicated decreased myocyte function during severe and resolved or resolving rejection. Cytoimmunological monitoring and quantitative birefringence measurements were diagnostically most useful in terms of sensitivity, specificity, and predictive value, when only data gathered before the first episode of acute rejection were considered. For cytoimmunological monitoring, diagnostic relevance was optimal when the data were expressed as relative proportions of activated lymphocytes. The quantitative birefringence measurements correlated best with analysis of the endomyocardial biopsy specimen when a cut off value of 1.25 was used. When both methods for diagnosing acute rejection were analysed together, no improvement in sensitivity (value 0.44) was found, but the specificity increased to 0.98 and the predictive value to about 0.80. It is concluded that cytoimmunological monitoring is a useful, non-invasive additional method for diagnosing the first period of acute rejection after heart transplantation and that quantitative birefringence measurements give valuable information on the extent of myocyte damage.

Effect of donor heart damage on survival after transplantation.

Evaluation of the functional condition of the heart prior to its removal from the donor or after transport to the recipient is difficult. Biopsies of the myocardium allow serial assessments to be made throughout this period, but suffer from the disadvantage that the average analysis of biopsies has only a tenuous connection with physiological function. Quantitative birefringence measurements (QBM), on the other hand, assess the ability of myocardial fibres to respond to ATP and calcium and have been shown to correlate well with measurements of cardiac function (P less than 0.001). A prospective study of myocardial biopsies before excision, after transport and again after transplantation, using quantitative birefringence measurement of biopsies of the heart has recently been completed. These studies have shown evidence of impaired myocardial function in 73 (43%) of 172 donor hearts studied prior to excision, with a further 27% showing significant deterioration during storage and transport to the recipient. Biopsy assessments therefore indicated that at the moment of implantation, only 30% of the donor hearts were normal. Functional assessment of the biopsies by QBM correlated with early clinical outcome of transplantation (P less than 0.001). Longer term follow-up of the recipients (up to 5 years) has shown that the mortality of recipients of hearts with impaired function before transplantation is significantly increased (44% of 120) compared with that of recipients of undamaged hearts (6% of 52, P less than 0.001).

Clinical validation of St. Thomas' Hospital cardioplegic solution No. 2 (Plegisol).

Recently, the St. Thomas' Hospital cardioplegic solution No. 2 (Plegisol) has become available commercially in the UK. In a series of patients (n = 28) undergoing open heart surgery for a variety of lesions, a clinical validation was performed. Preservation of myocardial contractility was assessed biophysically by quantitative birefringence measurements of myocardial biopsy samples (full thickness apical left ventricle and right ventricle) taken (1) prior to ischaemia, (2) at the end of ischaemia and (3) 10-15 min after reperfusion during cardiopulmonary bypass. In addition, serum CK-MB values were measured in samples taken throughout the operation and for 4 days postoperatively. Postoperative ECG traces (taken every 6 h for 48 h and then daily up to 7 days) were analysed to identify the occurrence of perioperative infarction. There were no hospital deaths. Chronotropic support was required in 5 of 28 patients (18%) for transient heart block. Low cardiac output did not occur postoperatively. Birefringence measurements in biopsy samples taken at the end of the ischaemic period (immediately prior to reperfusion) indicated an apparent left ventricular deterioration in myocardial contractility in 12 of 28 patients (43%) when compared to biopsies sampled prior to the ischaemic period. However, after 10-15 min of aerobic reperfusion, measurements indicated that myocardial contractility recovered to almost pre-ischaemic levels in the majority of patients. Thus, in 22 of 28 patients (79%), left ventricular deterioration did not occur in post-ischaemic biopsy samples when compared to the pre-ischaemic biopsies. Similarly, 21 of 28 patients (75%) had no deterioration of birefringence values in right ventricular biopsies.(ABSTRACT TRUNCATED AT 250 WORDS)

Biopsy assessment of fifty hearts during transplantation

The aim of the study was to evaluate myocardial protection during the transportation of human donor hearts stored at 4 degrees C in crystalloid cardioplegic solution. Biopsies were performed on 50 donor hearts before excision and at four subsequent time intervals during transplantation. Quantitative birefringence measurements on cryostat sections of the left ventricular biopsy specimens were used to assess myocardial function. Fifteen donor hearts had poor birefringence assessments before excision, and 67% received inotropic support after implantation; 22 deteriorated during transportation, and 50% received inotropic support; 13 were unchanged throughout the procedure, and none required inotropic support. This study demonstrates the need for improved protection of donor hearts during transportation and recommends the use of rigorous criteria in the selection of these hearts.

Numerical evaluation of extinction angle and birefringence in various directions as a function of velocity gradient

Despite the limitations of the theory of Peterlin and Stuart it still seems to be the best theory for interpreting quantitative flow birefringence measurements performed on solutions of rigid spheroidal particles. First, we repeat the numerical evaluation of the extinction angleX and the orientation factorfz corresponding to conventional observations realized in a Couette cell, but the values are now given with a higher degree of accuracy and the range of variation of the parameters has been extended. Furthermore, using the same theory, we extend our calculations to other directions of observation. This should contribute towards avoiding false interpretations of birefringence measurements in these directions (as has sometimes occurred). Finally, we present measurements taken along the lines of flow and along the direction of the velocity gradient, thus including the principal situations met in the various cells used in flow birefringence measurements.

Myocardial protection and function assessed by quantitative birefringence measurements: S. Darracott-Cankovic, J. Chayen, M.V. Braimbridge, Cardiothoracic Research, The Rayne Institute, St. Thomas' Hospital, London SE1, U.K.

Myocardial protection and function assessed by quantitative birefringence measurements: S. Darracott-Cankovic, J. Chayen, M.V. Braimbridge, Cardiothoracic Research, The Rayne Institute, St. Thomas' Hospital, London SE1, U.K.

Cellular chemical indices of right ventricular protection in children.

Myocardial preservation during open heart surgery in children was studied using biopsies of the right ventricle taken at the beginning and end of a bypass with a Tru-Cut biopsy needle. Three assessments were used: (1) semi-quantitative cytochemical grading of Baker's acid haematein reaction, and the distribution of succinate dehydrogenase and myosin ATPase; (2) microdensitometry of the succinate dehydrogenase activity; and (3) quantitative birefringence measurements to assess the response of the myocardial fibres to ATP and calcium. For each assessment, the values at the beginning and end of the bypass were compared. In a series of 42 children, two died in low cardiac output and three others required substantial inotropic support. No patient showed deterioration in the overall cytochemical assessment. Two patients showed deterioration in birefringence, one died and one had low cardiac output. The remaining three patients who had post-operative problems had low birefringence values at the beginning of bypass. Thirty-two patients were used for the microdensitometric assays, one died and three required substantial inotropic support. Succinate dehydrogenase activity decreased significantly during bypass in only two patients. One of these died and the other required substantial inotropic support.