A robust modelling method was proposed to extract chromophore information in multi-layered skin tissue with spatially-resolved diffuse reflectance spectroscopy. Artificial neural network models trained with a pre-simulated database were first built to map geometric and optical parameters into diffuse reflectance spectra. Nine fitting parameters including chromophore concentrations and oxygen saturation were then determined by solving the inverse problem of fitting spectral measurements from three different parts of the skin. Compared to the Monte Carlo simulation accelerated by a graphics processing unit, the proposed modelling method not only reduced the computation time, but also achieved a better fitting performance.

Tumor antigen-specific cytotoxic T lymphocyte (CTL) is a promising agent for cancer therapy. Most solid tumors are characterized by increased interstitial fluid pressure (IFP) and dense collagen capsule, which form physical barriers to impede cancer treatment. However, it remains unclear how CTL-mediated anticancer response is affected at the presence of these obstacles. Using a microfluidic-based platform mimicking these obstacles, we investigated the migration characteristics and performance of anticancer response of CTLs targeting hepatic cancer cells via antigen-specific and allogeneic recognition. The device consisted of slit channels mimicking the narrow interstitial paths constrained by the fibrous capsule and increased IFP was simulated by applying hydrostatic pressure to the tumor center. We found that antigen-specificity of CTLs against the targeted cancer cells determined the cytotoxic efficacy of the CTLs but did not significantly affect the success rate in CTLs that attempted to infiltrate into the tumor center. When increased IFP was present in the tumor center, CTL recruitment to tumor peripheries was promoted but success of infiltration was hindered. our results highlight the importance of incorporating the physical characteristics of tumor interstitum into the development of CTL-based cancer immunotherapy. Tumor antigen-specific CD8 + cytotoxic T lymphocyte (CTL)-mediated killing of tumor cells has a crucial role in cancer immunotherapy 1. Success of CTL-mediated tumor rejection requires the recruitment, infiltration, and expansion of tumor antigen-specific CTLs in tumor interstitium-the fluidic and matrix compartments between vessels and tumor cells, and recognition and killing of the tumor cells by the CTLs 2. However, a large body of evidence indicates that tumor cells actively reprogram surrounding interstitium to restrict CTLs from interacting with the tumor cells 3. For example, many types of cancer upregulate endothelins signalling of tumor endothelium to impede CTLs infiltration in tumor 4,5 ; soluble mediators such as IL-10 and transforming growth factor β (TGF-β) secreted by either tumor cells or tumor-recruited Treg cells significantly suppress the cytotoxic function of CTLs 3. While a multitude of chemical factors employed by cancers to escape from anticancer immunity are disclosed 6 , an increasing interest has recently been gained in the physical barriers established by tumors in their interstitium, which also poses a significant challenge to CTLs to successfully contact the targeting cells 7,8. Direct delivery of immune cells into tumor interior via perfusion may be physically hindered by the increased vascular resistance imposed by the high compressive stress generated by tumor growth 9,10. The growth-induced solid stress is mainly contributed by the collagen network and space-taking molecules, such as hyaluronan, accumulated in the tumor interstitium 11. Strategies to improve the delivery of blood-borne therapeutic agents against tumor, including the anticancer immune cells, has ...

The capability to image the 3D distribution of melanin in human skin in vivo with absolute quantities and microscopic details will not only enable noninvasive histopathological diagnosis of melanin-related cutaneous disorders, but also make long term treatment assessment possible. In this paper, we demonstrate clinical in vivo imaging of the melanin distribution in human skin with absolute quantities on mass density and with microscopic details by using label-free third-harmonic-generation (THG) enhancement-ratio microscopy. As the dominant absorber in skin, melanin provides the strongest THG nonlinearity in human skin due to resonance enhancement. We show that the THG-enhancement-ratio (erTHG) parameter can be calibrated in vivo and can indicate the melanin mass density. With an unprecedented clinical imaging resolution, our study revealed erTHG-microscopy's unique capability for long-term treatment assessment and direct clinical observation of melanin's micro-distribution to shed light into the unknown pathway and regulation mechanism of melanosome transfer and translocation.

Skin consists of epidermis and dermis layers that have distinct optical properties. The quantification of skin optical properties is commonly achieved by modeling photon propagation in tissue using Monte Carlo (MC) simulations and iteratively fitting experimentally measured diffuse reflectance spectra. In order to speed up the inverse fitting process, time-consuming MC simulations have been replaced by artificial neural networks to quickly calculate reflectance spectra given tissue geometric and optical parameters. In this study the skin was modeled to consist of three layers and different scattering properties of the layers were considered. A new inverse fitting procedure was proposed to improve the extraction of chromophore-related information in the skin, including the hemoglobin concentration, oxygen saturation and melanin absorption. The performance of the new inverse fitting procedure was evaluated on 40 sets of simulated spectra. The results showed that the fitting procedure without knowing the epidermis thickness extracted chromophore information with accuracy similar to or better than fitting with known epidermis thickness, which is advantageous for practical applications due to simpler and more cost-effective instruments. In addition, the melanin volume fraction multiplied by the thickness of the melanin-containing epidermis layer was estimated more accurately than the melanin volume fraction itself. This product has the potential to provide a quantitative indicator of melanin absorption in the skin. In-vivo cuff occlusion experiments were conducted and skin optical properties extracted from the experiments were comparable to the results of previously reported in vivo studies. The results of the current study demonstrated the applicability of the proposed method to quantify the optical properties related to major chromophores in the skin, as well as scattering coefficients of the dermis. Therefore, it has the potential to be a useful tool for quantifying skin optical properties in vivo.

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