Flexible polarimetric probe for 3 × 3 Mueller matrix measurements of biological tissue.

Polarimetry is a noninvasive method that uses polarised light to assess biophysical characteristics of tissues. A series of incident polarisation states illuminates a biological sample, and analysis of sample-altered polarisation states enables polarimetric tissue assessment. The resultant information can, for example, help quantitatively differentiate healthy from pathologic tissue. However, most bio-polarimetric assessments are performed using free-space optics with bulky optical components. Extension to flexible fibre-based systems is clinically desirable, but is challenging due to polarisation-altering properties of optical fibres. Here, we propose a flexible fibre-based polarimetric solution, and describe its design, fabrication, calibration, and initial feasibility demonstration in ex vivo tissue. The design is based on a flexible fibre bundle of six multimode optical fibres, each terminated with a distal polariser that ensures pre-determined output polarisation states. The resultant probe enables linear 3 × 3 Mueller matrix characterization of distal tissue. Potential in vivo Mueller matrix polarimetric tissue examinations in various directly-inaccessible body cavities are envisioned.

Rapid wide-field Mueller matrix polarimetry imaging based on four photoelastic modulators with no moving parts.

A new polarimetry method is demonstrated to image the entire Mueller matrix of a turbid sample using four photoelastic modulators (PEMs) and a charge coupled device (CCD) camera, with no moving parts. Accurate wide-field imaging is enabled with a field-programmable gate array (FPGA) optical gating technique and an evolutionary algorithm (EA) that optimizes imaging times. This technique accurately and rapidly measured the Mueller matrices of air, polarization elements, and turbid phantoms. The system should prove advantageous for Mueller matrix analysis of turbid samples (e.g., biological tissues) over large fields of view, in less than a second.

Polarized light imaging in biomedicine: emerging Mueller matrix methodologies for bulk tissue assessment.

Polarized light point measurements and wide-field imaging have been studied for many years in an effort to develop accurate and information-rich tissue diagnostic methods. However, the extensive depolarization of polarized light in thick biological tissues has limited the success of these investigations. Recently, advances in technology and conceptual understanding have led to a significant resurgence of research activity in the promising field of bulk tissue polarimetry. In particular, with the advent of improved measurement, analysis, and interpretation methods, including Mueller matrix decomposition, new diagnostic avenues, such as quantification of microstructural anisotropy in bulk tissues, have been enabled. Further, novel technologies have improved the speed and the accuracy of polarimetric instruments for ex vivo and in vivo diagnostics. In this paper, we review some of the recent progress in tissue polarimetry, provide illustrative application examples, and offer an outlook to the future of polarized light imaging in bulk biological tissues.

Assessment of local structural disorders of the bladder wall in partial bladder outlet obstruction using polarized light imaging.

Partial bladder outlet obstruction causes prominent morphological changes in the bladder wall, which leads to bladder dysfunction. In this paper, we demonstrate that polarized light imaging can be used to identify the location of obstruction induced structural changes that other imaging modalities fail to detect. We induced 2-week and 6-week partial outlet obstruction in rats, harvested obstructed bladders, then measured their retardances while distended to high pressures and compared them to controls. Our results show that the retardance of the central part of the ventral side (above the ureters) closer to the urethra can be used as a potential metric of the distending bladder obstruction.

Rapid time-gated polarimetric Stokes imaging using photoelastic modulators.

We report a rapid time-gated full Stokes imaging approach without mechanically moving parts, which is well-suited for biomedical applications, using two photoelastic modulators (PEMs). A charge-coupled device (CCD) with microsecond time-gating capability was used to acquire the images. To synchronize the CCD with the PEMs, thus gaining signal-to-noise ratio advantage, a field programmable gate array was employed. After calibration, an evolutionary algorithm was used to select four time points from which the full Stokes vector can be recovered. Using the images taken by the camera at these four times (in ~80 ms), the images of the full Stokes vectors of different incident polarization states were accurately derived.

Detecting axial heterogeneity of birefringence in layered turbid media using polarized light imaging.

The structural anisotropy of biological tissues can be quantified using polarized light imaging in terms of birefringence; however, birefringence varies axially in anisotropic layered tissues. This may present ambiguity in result interpretation for techniques whose birefringence results are averaged over the sampling volume. To explore this issue, we extended the polarization sensitive Monte Carlo code to model bi-layered turbid media with varying uniaxial birefringence in the two layers. Our findings demonstrate that the asymmetry degree (ASD) between the off-diagonal Mueller matrix elements of heterogeneously birefringent samples is higher than the homogenously birefringent (uniaxial) samples with the same effective retardance (magnitude and orientation). We experimentally verified the validity of ASD as a birefringence heterogeneity measure by performing polarized light measurements of bi-layered elastic and scattering polyacrylamide phantoms.

Optical assessment of tissue anisotropy in ex vivo distended rat bladders.

Microstructural remodelling in epithelial layers of various hollow organs, including changes in tissue anisotropy, are known to occur under mechanical distension and during disease processes. In this paper, we analyze how bladder distension alters wall anisotropy using polarized light imaging (followed by Mueller matrix decomposition). Optical retardance values of different regions of normal rat bladders under different distension pressures are derived. Then optical coherence tomography is used to measure local bladder wall thicknesses, enabling the calculation of the tissue birefringence maps as a measure of the tissue anisotropy. Selected two-photon microscopy is also performed to better understand the compositional origins of the obtained anisotropy results. The dome region of the bladder shows maximum birefringence when the bladder is distended to high pressures, whereas the ventral remains roughly isotropic during distension. In addition, the average anisotropy direction is longitudinal, along the urethra to dome. The derived wall anisotropy trends are based on birefringence as an intrinsic property of the tissue organization independent of its thickness, to aid in understanding the structure-functions relation in healthy bladders. These new insights into the wall microstructure of ex vivo distending bladders may help improve the functionality of the artificially engineered bladder tissues.

Quantitative correlation between light depolarization and transport albedo of various porcine tissues.

We present a quantitative study of depolarization in biological tissues and correlate it with measured optical properties (reduced scattering and absorption coefficients). Polarized light imaging was used to examine optically thick samples of both isotropic (liver, kidney cortex, and brain) and anisotropic (cardiac muscle, loin muscle, and tendon) pig tissues in transmission and reflection geometries. Depolarization (total, linear, and circular), as derived from polar decomposition of the measured tissue Mueller matrix, was shown to be related to the measured optical properties. We observed that depolarization increases with the transport albedo for isotropic and anisotropic tissues, independent of measurement geometry. For anisotropic tissues, depolarization was higher compared to isotropic tissues of similar transport albedo, indicating birefringence-caused depolarization effects. For tissues with large transport albedos (greater than ~0.97), backscattering geometry was preferred over transmission due to its greater retention of light polarization; this was not the case for tissues with lower transport albedo. Preferential preservation of linearly polarized light over circularly polarized light was seen in all tissue types and all measurement geometries, implying the dominance of Rayleigh-like scattering. The tabulated polarization properties of different tissue types and their links to bulk optical properties should prove useful in future polarimetric tissue characterization and imaging studies.

Do different turbid media with matched bulk optical properties also exhibit similar polarization properties?

We here investigate polarimetric behavior of thick samples of porcine liver, Intralipid, and microsphere-based tissue phantoms whose absorption and scattering properties are matched. Using polarized light we measured reflection mode Mueller matrices and derived linear/circular/total depolarization rates, based on polar decomposition. According to our results, phantoms exhibit greater depolarization rates in the backscattering geometry than the liver sample. The enhanced tissue polarization preservation differs from previous reports of polarimetric transmission studies, with the likely cause of this difference being the angular dependence of the single-scattering phase function. Also, Intralipid approximated polarimetric liver behavior well, whereas the polystyrene phantoms did not.

Evolutionary algorithm for solving the inverse problem of finding the incident field on a high-aperture lens for generating a desired focused field.

Using a proposed evolutionary algorithm, we have solved the inverse problem of finding the incident field on a high-aperture lens for generating a desired focused field, for the first time (to our knowledge). Further, we have achieved the global solution to the problem using this novel algorithm.

Images of finite sized spherical particles in confocal and conventional microscopes when illuminated with arbitrary polarization.

We investigate the form of the image of a finite sized spherical particle in confocal and conventional microscopes when the illuminating light has an arbitrary polarization. In particular, we take the cases of radial and azimuthal polarizations and use the Mie theory to find the scattered field from differently sized particles for these cases. We present numerical results for the changes in the detected intensity when subresolution and resolvable spherical particles are illuminated with particular wavelengths and polarizations. Further, we find the limiting size of a particle for which it can be considered a point scatterer for a particular wavelength.