Special Section Guest Editorial - Tissue Polarimetry

Tissue polarimetry is an emerging area of technology development in tissue optics that is expected to lead to important research applications in biology and medicine. Traditional polarimetry is a mature field, but its application to thick tissues has been hindered by technical hurdles associated with multiple light scattering. However, light is intrinsically polarized; and many biological tissue components or molecules—such as collagen, muscle fibers, keratin, retina, and glucose—possess polarization properties. Further, biological scatterers—such as cell nuclei and mitochondria—alter light polarization upon each scattering event according to the geometric and optical parameters of the scatterers. Since incident polarized light is rapidly depolarized by scattering, polarization-sensitive detection of reflected or transmitted light selects only the early escaping photons and rejects multiply scattered light. Hence, polarized light offers a means of gating: reflected polarized photons interrogate superficial tissue layers, whereas transmitted polarized photons image the ballistic or quasi-ballistic region. Polarization can therefore provide novel contrast mechanisms for imaging, diagnosis, and sensing

Light backscattering polarization patterns from turbid media: theory and experiment

We present both experimental measurements and Monte-Carlo-based simulations of the diffusely backscattered intensity patterns that arise from illuminating a turbid medium with a polarized laser beam. It is rigorously shown that, because of axial symmetry of the system, only seven elements of the effective backscattering Mueller matrix are independent. A new numerical method that allows simultaneous calculation of all 16 elements of the two-dimensional Mueller matrix is used. To validate our method we compared calculations to measurements from a turbid medium that consisted of polystyrene spheres of different sizes and concentrations in deionized water. The experimental and numerical results are in excellent agreement

Real-time, closed-loop dual-wavelength optical polarimetry for glucose monitoring

The development of a real-time, dual-wavelength optical polarimetric system to ultimately probe the aqueous humor glucose concentrations as a means of noninvasive diabetic glucose monitoring is the long-term goal of this research. The key impact of the work is the development of an approach for the reduction of the time-variant corneal birefringence due to motion artifact, which is still a limiting factor preventing the realization of such a device. Our dual-wavelength approach utilizes real-time, closed-loop feedback that employs a classical three-term feedback controller and efficiently reduces the effect of motion artifact that appears as a common noise source for both wavelengths. In vitro results are shown for the open-loop system, and although the dual-wavelength system helps to reduce the noise, it is shown that closed-loop control is necessary to bring the noise down to a sufficient level for physiological monitoring. Specifically, in vitro measurement results with the closed-loop dual-wavelength approach demonstrate a sensitivity of 12.8 mg∕dl across the physiologic glucose range in the presence of time-variant test cell birefringence. Overall, it is shown that this polarimetric system has the potential to be used as a noninvasive measure of glucose for diabetes

Diffuse reflectance polarization images of turbid media affected by glucose

Diffuse reflectance polarimetry has been used to obtain images from tissue-simulating turbid media with different glucose concentrations to investigate the changes in the polarization patterns induced by glucose. The optical properties change as the concentration of glucose is varied in the medium. The polarization patterns are affected by the overall optical properties of the turbid medium under study. This approach has potential applications in actual biological tissues for rapidly determining glucose levels non-invasively

High Affinity Mannotetraose as an Alternative to Dextran in ConA Based Fluorescent Affinity Glucose Assay Due to Improved FRET Efficiency

Diabetes mellitus affects millions of people worldwide and requires that individuals tightly self-regulate their blood glucose levels to minimize the associated secondary complications. Continuous monitoring devices potentially offer patients a long-term means to tightly monitor their glucose levels. In recent years, fluorescent affinity sensors based on lectins (e.g., Concanavalin A (ConA)) have been implemented in such devices. Traditionally, these sensors pair the lectin with a multivalent ligand, like dextran, in order to develop a competitive binding assay that changes its fluorescent properties in response to the surrounding glucose concentrations. This work introduces a new type of fluorescent ligand for FRET-based assays in an attempt to improve the sensitivity of such assays. This ligand is rationally designed to present a core trimannose structure and a donor fluorophore in close proximity to one another. This design decreases the distance between the FRET donor and the FRET acceptors on ConA to maximize the FRET efficiency upon binding of the ligand to ConA. This work specifically compares the FRET efficiency and sensitivity of this new competing ligand with a traditional dextran ligand, showing that the new ligand has improved characteristics. This work also tested the long-term thermal stability of the assay based on this new competing ligand and displayed a MARD of less than 10% across the physiological range of glucose after 30 days incubation at 37 °C. Ultimately, this new type of fluorescent ligand has the potential to significantly improve the accuracy of continuous glucose monitoring devices based on the competitive binding sensing approach

Optimization of a Concanavalin A‑Based Glucose Sensor Using Fluorescence Anisotropy

To date, the dependent nature of the recognition and transduction mechanisms in optical glucose sensors based upon Concanavalin A (ConA) has tended to prevent the sensors’ full potential from being realized. In this paper, these mechanisms are independently optimized for a given assay configuration in order to decrease the predictive error of a ConA-based glucose sensor and to give a more accurate demonstration of its potential. To this end, we used fluorescence anisotropy as the transduction mechanism to determine the binding of ConA to 4 kDa FITC-dextran by measuring the change in the rotational correlation lifetime between the bound and unbound populations. By tracking the fluorescence anisotropy of this ligand, the ranges of ConA and 4 kDa FITC-dextran concentrations capable of being explored were not limited by the transduction mechanism. Using predetermined association constants, the binding responses to physiological glucose concentrations were predicted for different assay configurations, and experimentally collected fluorescence anisotropy data displayed the predicted trends for these assay configurations. From the experimental results, a calibration fit was generated for the optimized assay configuration to predict the glucose concentrations using the fluorescence anisotropy. This optimized assay displayed a mean standard error of prediction of 7.5 mg/dL (0–300 mg/dL), and 100% of the data points fell within clinically acceptable zones (A and B) upon the Clarke Error Grid Analysis. This indicates that, by independently optimizing the recognition and transduction mechanisms for the final assay configuration, the sensitivity of a competitive binding chemistry using ConA can be appropriately configured for continuous glucose monitoring applications

Rational Design of a Bisphenol A Aptamer Selective Surface-Enhanced Raman Scattering Nanoprobe

Surface-enhanced Raman scattering (SERS) optical nanoprobes offer a number of advantages for ultrasensitive analyte detection. These functionalized colloidal nanoparticles are a multifunctional assay component. providing a platform for conjugation to spectral tags, stabilizing polymers, and biorecognition elements such as aptamers or antibodies. We demonstrate the design and characterization of a SERS-active nanoprobe and investigate the nanoparticles’ biorecognition capabilities for use in a competitive binding assay. Specifically, the nanoprobe is designed for the quantification of bisphenol A (BPA) levels in the blood after human exposure to the toxin in food and beverage plastic packaging. The nanoprobes demonstrated specific affinity to a BPA aptamer with a dissociation constant <i>K</i>_d of 54 nM, and provided a dose-dependent SERS spectra with a limit of detection of 3 nM. Our conjugation approach shows the versatility of colloidal nanoparticles in assay development, acting as detectable spectral tagging elements and biologically active ligands concurrently

Self-Cleaning Membrane to Extend the Lifetime of an Implanted Glucose Biosensor

The lifetime and efficacy of a subcutaneously implanted glucose biosensor could be greatly improved by a self-cleaning membrane capable of periodic physical removal of adhered cells associated with the foreign body reaction. Previously, we reported a thermoresponsive double network nanocomposite (DNNC) membrane composed of poly­(<i>N</i>-isopropylacrylamide) (PNIPAAm) and embedded polysiloxane nanoparticles. When the membrane was thermally cycled above and below its volume phase transition temperature (VPTT, ∼33–35 °C), the associated deswelling and reswelling, respectively, led to in vitro cell release. Herein, this membrane design was tailored to meet the specific demands of a subcutaneously implanted glucose biosensor, and critical functional properties were assessed. First, <i>N</i>-vinylpyrrolidone (NVP) comonomer increased the VPTT to ∼38 °C so that the membrane would be swollen and thus more permeable to glucose in the “off-state” (i.e., no heating) while residing in the subcutaneous tissue (∼35 °C). Second, glucose diffusion kinetics though the DNNC membrane was experimentally measured in its deswollen and reswollen states. A cylindrical DNNC membrane with dimensions considered suitable for implantation (1.5 × 5 mm, diameter × length) was used to model the glucose diffusion lag time. In addition, the DNNC cylinder was used to observe dimensional changes associated with deswelling and reswelling. Noncytotoxicity was confirmed and self-cleaning was assessed in vitro in terms of thermally driven cell release to confirm the potential of the DNNC membrane to control biofouling