SignificanceAlthough spatial frequency domain imaging (SFDI) has been well characterized under diffuse optical conditions, tissue measurements made outside the diffuse regime can provide new diagnostic information. Before such measurements can become clinically relevant, however, the behavior of sub-diffuse SFDI and its effect on the accuracy of derived tissue parameters must be assessed.AimWe aim to characterize the impact that both the assumed scattering phase function (SPF) and the polarization state of the illumination light source have on the accuracy of SFDI-derived optical properties when operating under diffuse or sub-diffuse conditions, respectively.ApproachThrough the use of a set of well-characterized optical phantoms, SFDI accuracy was assessed at four wavelengths (395, 545, 625, and 850 nm) and two different spatial frequencies (0.3 and 1.0 mm−1), which provided a broad range of diffuse and sub-diffuse conditions, using three different SPFs. To determine the effects of polarization, the SFDI accuracy was assessed using both unpolarized and cross-polarized illumination.ResultsIt was found that the assumed SPF has a direct and significant impact on the accuracy of the SFDI-derived optical properties, with the best choice of SPF being dictated by the polarization state. As unpolarized SFDI retains the sub-diffuse portion of the signal, optical properties were found to be more accurate when using the full SPF that includes forward and backscattering components. By contrast, cross-polarized SFDI yielded accurate optical properties when using a forward-scattering SPF, matching the behavior of cross-polarization to attenuate the immediate backscattering of sub-diffuse reflectance. Using the correct pairings of SPF and polarization enabled using a reflectance standard, instead of a more subjective phantom, as the reference measurement.ConclusionsThese results provide the foundation for a more thorough understanding of SFDI and enable new applications of this technology in which sub-diffuse conditions dominate (e.g., μa≮μs′) or high spatial frequencies are required.
Current approaches of creating optical phantoms cannot accurately capture the wavelength-dependent properties found in tissue. To address this, we developed a method of producing solid, inorganic phantoms whose wavelength-dependent optical properties can be fit to those of tissue over 370 to 950 nm through the combination of up to twenty different absorbing and scattering pigments. Using this approach, we were able to create and validate spectral phantoms closely matching the optical properties of muscle and nerve tissue, the diffuse reflectance of pale and melanistic skin, and the chromophore concentrations of a computational skin model with varying levels of oxygen saturation.
SignificanceCurrent methods of producing optical phantoms are incapable of accurately capturing the wavelength-dependent properties of tissue critical for many optical modalities.AimWe aim to introduce a method of producing solid, inorganic phantoms whose wavelength-dependent optical properties can be matched to those of tissue over the wavelength range of 370 to 950 nm.ApproachThe concentration-dependent optical properties of 20 pigments were characterized and used to determine combinations that result in optimal fits compared to the target properties over the full spectrum. Phantoms matching the optical properties of muscle and nerve, the diffuse reflectance of pale and melanistic skin, and the chromophore concentrations of a computational skin model with varying oxygen saturation (StO2) were made with this method.ResultsBoth optical property phantoms were found to accurately mimic their respective tissues’ absorption and scattering properties across the entire spectrum. The diffuse reflectance phantoms were able to closely approximate skin reflectance regardless of skin type. All three computational skin phantoms were found to have emulated chromophore concentrations close to the model, with an average percent error for the StO2 of 4.31%.ConclusionsThis multipigment phantom platform represents a powerful tool for creating spectrally accurate tissue phantoms, which should increase the availability of standards for many optical techniques.
Coupling of surface-enhanced Raman spectroscopy (SERS) with the coffee ring effect can overcome the poor reproducibility typically seen when using SERS. In this study, we developed a nitrocellulose membrane paper-based substrate for coffee ring enhanced SERS, which was highly hydrophobic and produced consistent coffee rings. After optimization of solution parameters including gold nanoparticle concentration and solvent, this platform demonstrated high enhancement and low variability using Malachite Green Isothiocyanate and Moraxella catarrhalis. This substrate has the potential to increase the usability and implementation of SERS by overcoming intrinsic limitations and is more accessible than current substrates.
Iatrogenic nerve injuries often occur during surgical procedures yielding high morbidity. Numerous optical techniques are being investigated to provide nerve visualization and monitoring to both avoid and detect nerve injury. However, there is insufficient VIS-IR optical properties data for nerve and surrounding tissues, to facilitate needed advancements in the biophotonic approaches being developed for deployment. To fill this gap, we determined the absorption and reduced scattering coefficients of ex vivo nerve, muscle, tendon, and adipose tissues from four rats and human cadavers between 350-2500nm. The results provide valuable insights for optimizing the optical contrast of nerves for improved surgical guidance.
Current methods of photodynamic therapy dosimetry are limited in their spatial and temporal resolutions. To address this, we have designed a system that combines therapeutic illumination with both fluorescence and spatial frequency domain imaging (SFDI). Fluorescence imaging during treatment provides information on the photosensitizer distribution and photobleaching rate while SFDI allows for the approximation of fluence, oxygenation, and tissue-corrected photosensitizer concentrations, but requires the treatment be paused. To minimize these interruptions, the single snapshot of optical properties approach was used which requires only a single image frame. A prototype of the system has been built and validated using calibrated phantoms.
One of the promising therapeutic effects of photobiomodulation is the ability to modulate the redox homeostasis of a cell. While this effect may be beneficial in treating chronic or infected wounds by boosting the antimicrobial capabilities of neutrophils, it is poorly understood how the redox state of bacteria is altered due to exposure to the red or near-infrared light. To address this, we present the effects of 690 nm light on the response of S. aureus to exogenous hydrogen peroxide at a range of different optical parameters including irradiance and fluence.
The increased incidence of antibiotic-resistant gram-positive bacteria, like methicillin-resistant S. aureus (MRSA), necessitates treatments that eliminate the potential of developing further resistance. Antimicrobial photodynamic therapy (aPDT) has shown promise as gram-positive infections can be specifically photosensitized by inducing the accumulation of coproporphyrin III (CPIII) through the administration of VU0038882 (‘882), a small-molecule activator of coproporphyrinogen oxidase, and delta-aminolevulinic acid hydrochloride (ALA). While the phototoxic effects of CPIII are most pronounced when stimulated with 395nm light, corresponding to its Soret absorption-band, the high absorption of the skin at that wavelength reduces the efficacy in vivo by three orders of magnitude as compared to in vitro. Although the issue of light penetrance can be mitigated by using red-shifted wavelengths targeting the Q-bands of CPIII (λpeak=498/530/565/619nm), the efficiency of cytotoxic reactive oxygen species (ROS) production and bacterial killing drastically reduces. Though this inefficiency can be partially overcome through an increased light dose, photoinactivation of CPIII and oxygen depletion limits this process to a maximum effective light dose. To overcome these limitations and improve the overall efficacy of CPIII-targeted aPDT, we designed and built a novel multi-LED light source and explored the effect of simultaneously targeting the Soret-band and Q-bands. We present that lower radiant exposures of blue light in conjunction with a higher exposure of green or red light increases the amount of bacterial killing by 1 to 3 logs in vitro as compared to either treatment alone. This enhancement is expected to increase when utilized in vivo due to differences in penetrance.
This work focuses on pulsed terahertz imaging for the application of surgical margin assessment of breast cancer. Various phantom tissue types and orientations are tested here to refine imaging methodology that can detect breast cancer up to 0.5-1.0 mm from the edge of the sample. The depth of the cancer within the sample is estimated using time of flight analysis of the reflected peaks in the pulsed time domain signal. Breast tissue phantoms have been designed to resemble fresh infiltrating ductal carcinoma, fibroglandular tissue, and fatty tissue of the breast to accomplish this work.
The goal of this work was to develop phantoms that match the refractive indices and absorption coefficients between 0.15 and 2.0 THz of the freshly excised tissues commonly found in breast tumors. Since a breast cancer tumor can contain fibrous and fatty tissues alongside the cancerous tissues, a phantom had to be developed for each. In order to match the desired properties of the tissues, oil in water emulsions were solidified using the proven phantom component TX151. The properties of each potential phantom were verified through THz time-domain spectroscopy on a TPS Spectra 3000. Using this method, phantoms for fibrous and cancerous tissue were successfully developed while a commercially available material was found which matched the optical properties of fatty tissue.
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