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The dating of organic findings is a fundamental task for many scientific fields. Radiocarbon dating is currently the most commonly used method. For wood, dendrochronology is another state‐of‐the‐art method. Both methods suffer from systematic restrictions, leading to samples that have not yet been able to be dated. Molecular changes over time are reported for many materials under different preservation conditions. Many of them are intrinsically monotonous. These monotonous molecular decay (MD) patterns can be understood as clocks that start at the time when a given molecule was formed. Factors that influence these clocks include input material composition and preservation conditions. Different wood species, degrees of pyrolysis, and pretreatments lead to different prediction models. Preservation conditions might change the speed of a given clock and lead to different prediction models. Currently published models for predicting the age of wood, paper, and parchment depend on infrared spectroscopy. In contrast to radiocarbon dating, dating via MD does not comprise a single methodology. Some clocks may deliver less precise results than the others. Ultimately, developing a completely different, new dating strategy‐such as MD dating–will help to bring to light a treasure trove of information hidden in the darkness of organic findings., This review focuses on molecular changes over time. Having been discussed since the 1960s, the variety of molecular clocks includes the change in amino acids in the primary structure of a protein over time. Molecular decay (MD) has a mainly monotonous function, a prerequisite to serving as such a clock. In addition, such clocks need a defined starting point and a defined decay function. In contrast to the decay of alpha and beta emitters, MD is influenced by several environmental factors indicated by preservation conditions. The MD method has the potential to be used for different organic materials and different scientific fields.

Background Exogenous materials may be encountered in skin biopsies as contamination. Contamination may occur during the biopsy procedure in the clinic or during tissue processing in the laboratory. Although the experienced pathologist may often instinctively ignore clear examples of contamination, such tissue may be a source of confusion for young pathologists and those in training. Foreign materials can typically be recognized as exogenous by morphology, polarizability, and the presence or absence of a tissue reaction, but we have rarely encountered situations in which exogenous materials could be misinterpreted as organisms, either by their morphologic appearance or staining properties. Methods Exogenous materials used during skin biopsy and grossing were embedded in a nonhuman tissue scaffold and prepared into histologic slides. Hematoxylin and eosin (H&E), periodic acid-Schiff with diastase, and Grocott methenamine silver stains were performed, and each material was evaluated under polarized microscopy. Results Exogenous materials were divided into the following 3 categories with shared morphologic appearances and staining properties: suture materials, plant-based materials, and synthetic materials. Conclusion We present a comprehensive characterization of the morphologic and cytochemical staining properties of multiple exogenous materials that may contaminate a skin biopsy. This characterization will aid the pathologist by providing a mechanism to identify potential contaminants in skin biopsies.

A soft body area sensor network presents a promising direction in wearable devices to integrate on-body sensors for physiological signal monitoring and flexible printed circuit boards (FPCBs) for signal conditioning/readout and wireless transmission. However, its realization currently relies on various sophisticated fabrication approaches such as lithography or direct printing on a carrier substrate before attaching to the body. Here, we report a universal fabrication scheme to enable printing and room-temperature sintering of the metal nanoparticle on paper/fabric for FPCBs and directly on the human skin for on-body sensors with a novel sintering aid layer. Consisting of polyvinyl alcohol (PVA) paste and nanoadditives in the water, the sintering aid layer reduces the sintering temperature. Together with the significantly decreased surface roughness, it allows for the integration of a submicron-thick conductive pattern with enhanced electromechanical performance. Various on-body sensors integrated with an FPCB to detect health conditions illustrate a system-level example.

Significance: The spatial organization of collagen fibers has been used as a biomarker for assessing injury and disease progression. However, quantifying this organization for complex structures is challenging. Aim: To quantify and classify complex collagen fiber organizations. Approach: Using quantitative second-harmonic generation (SHG) microscopy, we show that collagen-fiber orientation can be viewed as pseudovector fields. Subsequently, we analyze them using fluid mechanic metrics, such as energy U, enstrophy E, and tortuosity tau. Results: We show that metrics used in fluid mechanics for analyzing fluid flow can be adapted to analyze complex collagen fiber organization. As examples, we consider SHG images of collagenous tissue for straight, wavy, and circular fiber structures. Conclusions: The results of this study show the utility of the chosen metrics to distinguish diverse and complex collagen organizations. We find that the distribution of values for E and U increases with collagen fiber disorganization, where they divide between low and high values corresponding to uniformly aligned fibers and disorganized collagen fibers, respectively. We also confirm that the values of tau cluster around 1 when the fibers are straight, and the range increases up to 1.5 when wavier fibers are present. (C) The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Published version

Significance: Sub-diffuse optical properties may serve as useful cancer biomarkers, and wide-field heatmaps of these properties could aid physicians in identifying cancerous tissue. Sub-diffuse spatial frequency domain imaging (sd-SFDI) can reveal such wide-field maps, but the current time cost of experimentally validated methods for rendering these heatmaps precludes this technology from potential real-time applications. Aim: Our study renders heatmaps of sub-diffuse optical properties from experimental sd-SFDI images in real time and reports these properties for cancerous and normal skin tissue subtypes. Approach: A phase function sampling method was used to simulate sd-SFDI spectra over a wide range of optical properties. A machine learning model trained on these simulations and tested on tissue phantoms was used to render sub-diffuse optical property heatmaps from sd-SFDI images of cancerous and normal skin tissue. Results: The model accurately rendered heatmaps from experimental sd-SFDI images in real time. In addition, heatmaps of a small number of tissue samples are presented to inform hypotheses on sub-diffuse optical property differences across skin tissue subtypes. Conclusion: These results bring the overall process of sd-SFDI a fundamental step closer to real-time speeds and set a foundation for future real-time medical applications of sd-SFDI such as image guided surgery.

Significance: Artificial skin (AS) is widely used in dermatology, pharmacology, and toxicology, and has great potential in transplant medicine, burn wound care, and chronic wound treatment. There is a great demand for high-quality AS product and a non-invasive detection method is highly desirable. Aim: To quantify the constructure parameters (i.e., thickness and surface roughness) of AS samples in the culture cycle and explore the growth regularities using optical coherent tomography (OCT). Approach: An adaptive interface detection algorithm is developed to recognize surface points in each A-scan, offering a rapid method to calculate parameters without constructing OCT B-scan pictures and further achieving realizing real-time quantification of AS thickness and surface roughness. Experiments on standard roughness plates and H&E-staining microscopy were performed as a verification. Results: As applied on the whole cycle of AS culture, our method’s results show that during the air–liquid culture, the surface roughness of the skin first decreases and then exhibits an increase, which implies coincidence with the degree of keratinization under a microscope. And normal and typical abnormal samples can be differentiated by thickness and roughness parameters during the culture cycle. Conclusions: The adaptive interface detection algorithm is suitable for high-sensitivity, fast detection, and quantification of the interface with layered characteristic tissues, and can be used for non-destructive detection of the growth regularity of AS sample thickness and roughness during the culture cycle.

Significance: Spatial frequency domain imaging (SFDI) is a wide-field imaging technique that provides quantitative maps of tissue optical properties. We describe a compact SFDI imager that employs a multispectral compound-eye camera. This design enables simultaneous image acquisition at multiple wavelengths. Such a device has potential for application for quantitative evaluation of superficial tissues by nonspecialists in low-resource settings. Aim: The aim of this work was to develop a compact SFDI imager for widefield imaging of in-vivo tissue optical properties and verify its ability to measure optical properties of tissue-simulating phantoms and in a preclinical model of burn wounds. Approach: This compound-eye imager was constructed using a CMOS sensor subdivided into multiple regions, each having a bandpass filter and objective lens. The ability of the instrument to image optical properties was compared with (1) a commercial SFDI imager and (2) a laboratory-based system. Initial validation of ability to accurately characterize optical properties was performed using a tissue-simulating optical phantom. It was then applied to an established murine model of thermal contact burn severity. In-vivo measurements of the optical properties of rat skin were performed before and after the application of burns. Histology was used to verify burn severity. Results: Measurements of the tissue-simulating phantom optical properties made using the compound-eye imager agree with measurements made using the two comparison SFDI devices. For the murine burn model, the burns showed a decrease in the reduced scattering coefficient at all measurement wavelengths compared with preburn measurements at the same locations. This is consistent with previously reported changes in scattering that occur in full-thickness burns. Conclusion: We demonstrate the potential for SFDI to be translated into compact form factor using a compound-eye camera that is capable of obtaining multiple wavelengths channels simultaneously.

Significance: Collagen is the most abundant protein in vertebrates and is found in tissues that regularly experience tension, compression, and shear forces. However, the underlying mechanism of collagen fibril formation and remodeling is poorly understood. Aim: We explore how a collagen monomer is visualized using fluorescence microscopy and how its spatial orientation is determined. Defining the orientation of collagen monomers is not a trivial problem, as the monomer has a weak contrast and is relatively small. It is possible to attach fluorescence tags for contrast, but the size is still a problem for detecting orientation using fluorescence microscopy. Approach: We present two methods for detecting a monomer and classifying its orientation. A modified Gabor filter set and an automatic classifier trained by convolutional neural network based on a synthetic dataset were used. Results: By evaluating the performance of these two approaches with synthetic and experimental data, our results show that it is possible to determine the location and orientation with an error of ∼37 deg of a single monomer with fluorescence microscopy. Conclusions: These findings can contribute to our understanding of collagen monomers interaction with collagen fibrils surface during fibril formation and remodeling.

Significance: The Mueller matrix decomposition method is widely used for the analysis of biological samples. However, its presumed sequential appearance of the basic optical effects (e.g., dichroism, retardance, and depolarization) limits its accuracy and application. Aim: An approach is proposed for detecting and classifying human melanoma and non-melanoma skin cancer lesions based on the characteristics of the Mueller matrix elements and a random forest (RF) algorithm. Approach: In the proposal technique, 669 data points corresponding to the 16 elements of the Mueller matrices obtained from 32 tissue samples with squamous cell carcinoma (SCC), basal cell carcinoma (BCC), melanoma, and normal features are input into an RF classifier as predictors. Results: The results show that the proposed model yields an average precision of 93%. Furthermore, the classification results show that for biological tissues, the circular polarization properties (i.e., elements m44, m34, m24, and m14 of the Mueller matrix) dominate the linear polarization properties (i.e., elements m13, m31, m22, and m41 of the Mueller matrix) in determining the classification outcome of the trained classifier. Conclusions: Overall, our study provides a simple, accurate, and cost-effective solution for developing a technique for classification and diagnosis of human skin cancer.

In this research, collagen hydrolysates with different average molecular weights ( Mn ¯ ) from leather collagen were chosen as raw materials. Five environmental-friendly sizing agents (SA) were prepared by cross-linking collagen hydrolysates with glycol diglycidyl ether (GDE) and further grafting them with butyl acrylate (BA) and styrene (St). Then the compound sizing agents (SGDESA-x, x = 1, 2, 3 and 4) were obtained by simple physical mixing of GDESA and starch. The surface sizing performance of GDESA and compound sizing agents were studied. The research result shows that both physical, mechanical properties and water resistance of the corrugated paper coated by GDESA were significantly improved when Mn ¯ of collagen hydrolysate was about 10,000, and its emulsion exhibited robust stability in long standing time. Furthermore, when the SGDESA-2 was used as a sizing agent, the coated corrugated paper exhibited strong water resistance, good physical and mechanical properties even after refolded for 20 times.

As human beings have been consistently exposed to bisphenol A (BPA) and bisphenol S (BPS) derived from various products, the intake of BPS/BPA to humans has been extensively studied. However, using conventional biological matrices such as urine, blood, or dissected skin to detect BPS/BPA in the human body system requires longer exposure time to them, hardly defines the pollutant source of the accumulated BPS/BPA, and is often invasive. Herein, our new approach i.e. fingerprint analysis quantitatively confirms the transfer of BPS/BPA from receipts (specific pollution source) to human skin only within receipt-handling of “20 s”. When receipts (fingertip region size; ~1 cm2) containing 100–300 μg of BPS or BPA are handled, 20–40 μg fingerprint-1 of BPS or BPA is transferred to human skin (fingertip). This transferred amount of BPS/BPA can still be toxic according to the toxicity test using water fleas. As a visual evidence, a fingerprint map that matches the distribution of the absorbed BPS/BPA is developed using a mass spectrometry imaging tool. This is the first study to analyze fingerprints to determine the incorporation mechanism of emerging pollutants. This study provides an efficient and non-invasive environmental forensic tool to analyze amounts and sources of hazardous substances.

Objectives Bisphenol A (BPA) is the most used colour developer in thermal paper for cashiers receipts, labels, and tickets. BPA can migrate onto the skin and be absorbed when handling these papers. BPA is a known endocrine disruptor and is therefore being replaced in thermal paper by some alternatives such as Bisphenol S (BPS), D-8, and Pergafast 201® (PF201). To our knowledge, no studies have characterized skin permeation of these BPA alternatives. Methods We measured/characterized skin absorption for BPA, BPS, D-8, and PF201 through ex vivo human skin using flow-through diffusion cells according to OECD guideline 428. Skin samples were 7–12 per test substance from three different skin donors. Skin metabolism was studied for BPA. Dermal absorption was expressed as the amount of the BPA alternatives in the receptor fluid over applied dose in percent (%). Results The absorbed dose after 24 h of exposure was 25% for BPA, 17% for D-8, 0.4% for BPS, and Conclusions Skin absorption was in decreasing order: BPA > D-8 >> BPS > PF201. These results are in agreement with their log Kow and molecular weights. We provided here the necessary data to estimate the extent of skin absorption of BPA analogues, which is a necessary step in risk assessment, and ultimately evaluate public health risks posed by D-8, BPS, and PF201.

Biomarkers in sweat are a largely untapped source of health information. Most of the currently available sweat harvesting and testing devices are incapable of operating under low-sweat rates such as those experienced by humans at rest. Here we analyze the in vitro and in vivo sampling of sweat through osmosis via the use of a hydrogel interfaced with the skin, without need for active perspiration. The hydrogel also interfaces with paper-based microfluidics to transport the fluid via capillary forces toward a testing zone and then evaporation pad. We show that the hydrogel solute content and area of the evaporation pad regulate the long-term extraction of sweat and its associated biomarkers. The results indicate that the platform can sample biomarkers from a model skin system continuously for approximately 12 h. On-skin testing of the platform on both resting and exercising human subjects confirms that it can sample sweat lactate directly from the surface of skin. The results highlight that lactate in sweat increases with exercise and as a direct result of muscle activity. Implementation of such new principles for sweat fluid harvesting and management via wearable patch devices can contribute toward the advancement of next generation wearables.

Significance Rehabilitation through facial prostheses' main goal is to aid individual's social reintegration as well as improving their quality of life. However, this treatment is not yet widely available in Brazil due to the lack of specialized clinics and the cost associated with the high number of necessary medical appointments until the final result. One of the steps in the process consists of measuring skin color, which is observer-dependent and may suffer from the effect of metamerism. Aim The methodology of our work aims to obtain a standard between different devices and greater fidelity to the color seen in person in order to reduce face-to-face iterations, reduce costs, and ensure better final results. Approach A physical device and a computer program were improved from previous projects. The changes included implementing the Thin-Plate Spline 3D algorithm for color calibration, in addition to an optional non-uniform illumination correction in the process. We also aim to improve the project's accessibility using a colorimeter. The methodology and the algorithms were both compared to readings from direct skin measurements as well as color references. Results After processing, the ΔEab* metric between images from the same segments is taken with different cameras and conditions of illumination decreased from 18.81 ± 4.85 to 4.85 ± 1.72. In addition, when the images were compared to colorimetric readings of the skin, the difference went from 14.93 ± 4.11 to 5.85 ± 1.61. It was also observed that using a less expensive device did not impact the readings. The project is open source and available at Github. Conclusions The results demonstrate the possibility of applying the methodology to assist in the manufacturing of facial prostheses to decrease the total number of consultations, in addition to providing greater reliability of the final result.

Significance: Terahertz (THz) radiation has demonstrated a great potential in biomedical applications over the past three decades, mainly due to its non-invasive and label-free nature. Among all biological specimens, skin tissue is an optimal sample for the application of THz-based methods because it allows for overcoming some intrinsic limitations of the technique, such as a small penetration depth (0.1 to 0.3 mm for the skin, on average). Aim: We summarize the modern research results achieved when THz technology was applied to the skin, considering applications in both imaging/detection and treatment/modulation of the skin constituents. Approach: We perform a review of literature and analyze the recent research achievements in THz applications for skin diagnosis and investigation. Results: The reviewed results demonstrate the possibilities of THz spectroscopy and imaging, both pulsed and continuous, for diagnosis of skin melanoma and non-melanoma cancer, dysplasia, scars, and diabetic condition, mainly based on the analysis of THz optical properties. The possibility of modulating cell activity and treatment of various diseases by THz-wave exposure is shown as well. Conclusions: The rapid development of THz technologies and the obtained research results for skin tissue highlight the potential of THz waves as a research and therapeutic instrument. The perspectives on the use of THz radiation are related to both non-invasive diagnostics and stimulation and control of different processes in a living skin tissue for regeneration and cancer treatment.

Significance: Spatial frequency domain imaging (SFDI), a noncontact wide-field imaging technique using patterned illumination with multiple wavelengths, has been used to quantitatively measure structural and functional parameters of in vivo tissue. Using SFDI in a porcine model, we previously found that scattering changes in skin could potentially be used to noninvasively assess burn severity and monitor wound healing. Translating these findings to human subjects necessitates a better understanding of the variation in “baseline” human skin scattering properties across skin types and anatomical locations. Aim: Using SFDI, we aim to characterize the variation in the reduced scattering coefficient (μs′) for skin across a range of pigmentation and anatomic sites (including common burn locations) for normal human subjects. These measurements are expected to characterize baseline human skin properties to inform our use of SFDI for clinical burn severity and wound healing assessments. Approach: SFDI was used to measure μs′ in the visible- and near-infrared regime (471 to 851 nm) in 15 subjects at 10 anatomical locations. Subjects varied in age, gender, and Fitzpatrick skin type. Results: For all anatomical locations, the coefficient of variation in measured μs′ decreased with increasing wavelength. High intersubject variation in μs′ at visible wavelengths coincided with large values of the melanin extinction coefficient at those wavelengths. At 851 nm, where intersubject variation in μs′ was smallest for all anatomical locations and absorption from melanin is minimal, significant intrasubject differences in μs′ were observed at the different anatomical locations. Conclusions: Our study is the first report of wide-field mapping of human skin scattering properties across multiple skin types and anatomical locations using SFDI. Measured μs′ values varied notably between skin types at wavelengths where absorption from melanin was prominent. Additionally, μs′ varied considerably across different anatomical locations at 851 nm, where the confounding effects from melanin absorption are minimized.

Significance: Oxygenation is one of the skin tissue physiological properties to follow for patient care management. Furthermore, long-term monitoring of such parameters is needed at the patient bed as well as outside the hospital. Diffuse reflectance spectroscopy has been widely used for this purpose. Aim: The aim of the study is to propose a low-cost system for the long-term measurement of skin physiological parameters in contact. Approach: We have developed a low-cost, wearable, CMOS-based device. We propose an original method for processing diffuse reflectance data to calculate the tissue oxygen saturation (StO2). Results: We tested the device for the assessment of tissue oxygenation during a first-in-human clinical trial that took place at the Grenoble University Hospital France. Conclusions: The results of this clinical trial show a good accordance between our sensor and commercial devices used a reference.

Significance: Hemoglobin oxygen saturation and red blood cell (RBC) tissue fraction are important parameters when assessing microvascular status. Functional information can be attained using temporally resolved measurements performed during stimulus–response protocols. Pointwise assessments can currently be conducted with probe-based systems. However, snapshot multispectral imaging (MSI) can be used for spatial–temporal measurements. Aim: To validate if hemoglobin oxygen saturation and RBC tissue fraction can be quantified using a snapshot MSI system and an inverse Monte Carlo algorithm. Approach: Skin tissue measurements from the MSI system were compared to those from a validated probe-based system during arterial and venous occlusion provocation on 24 subjects in the wavelength interval 450 to 650 nm, to evaluate a wide range of hemoglobin oxygen saturation and RBC tissue fraction levels. Results: Arterial occlusion results show a mean linear regression R2 = 0.958 for hemoglobin oxygen saturation. Comparing relative RBC tissue fraction during venous occlusion results in R2 = 0.925. The MSI system shows larger dynamic changes than the reference system, which might be explained by a deeper sampling including more capacitance vessels. Conclusions: The snapshot MSI system estimates hemoglobin oxygen saturation and RBC tissue fraction in skin microcirculation showing a high correlation (R2 > 0.9 in most subjects) with those measured by the reference method. Funding: Swedens Innovation Agency VINNOVA via the programs Swelife and MedTech4Health [2017-01435, 2019-01522]; CENIIT research organization within Linkoping University [11.02]

Significance: Management of skin cancer worldwide is often a challenge of scale, in that the number of potential cases presented outweighs the resources available to detect and treat skin cancer. Aim: This project aims to develop a polarimetry probe to create an accessible skin cancer detection tool. Approach: An optical probe was developed to perform bulk tissue Stokes polarimetry, a technique in which a laser of known polarization illuminates a target, and the altered polarization state of the backscattered light is measured. Typically, measuring a polarization state requires four sequential measurements with different orientations of polarization filters; however, this probe contains four spatially separated detectors to take four measurements in one shot. The probe was designed to perform at a lower cost and higher speed than conventional polarimetry methods. The probe uses photodiodes and linear and circular film polarizing filters as detectors, and a low-coherence laser diode as its illumination source. The probe design takes advantage of the statistical uniformity of the polarization speckle field formed at the detection area. Results: Tests of each probe component, and the complete system put together, were performed to evaluate error and confirm the probe’s performance despite its low-cost components. This probe’s potential is demonstrated in a pilot clinical study on 71 skin lesions. The degree of polarization was found to be a factor by which malignant melanoma could be separated from other types of skin lesions.

Pencils and papers are ubiquitous in our society and have been widely used for writing and drawing, because they are easy to use, low-cost, widely accessible, and disposable. However, their applications in emerging skin-interfaced health monitoring and interventions are still not well explored. Herein, we report a variety of pencil–paper-based on-skin electronic devices, including biophysical (temperature, biopotential) sensors, sweat biochemical (pH, uric acid, glucose) sensors, thermal stimulators, and humidity energy harvesters. Among these devices, pencil-drawn graphite patterns (or combined with other compounds) serve as conductive traces and sensing electrodes, and office-copy papers work as flexible supporting substrates. The enabled devices can perform real-time, continuous, and high-fidelity monitoring of a range of vital biophysical and biochemical signals from human bodies, including skin temperatures, electrocardiograms, electromyograms, alpha, beta, and theta rhythms, instantaneous heart rates, respiratory rates, and sweat pH, uric acid, and glucose, as well as deliver programmed thermal stimulations. Notably, the qualities of recorded signals are comparable to those measured with conventional methods. Moreover, humidity energy harvesters are prepared by creating a gradient distribution of oxygen-containing groups on office-copy papers between pencil-drawn electrodes. One single-unit device (0.87 cm(2)) can generate a sustained voltage of up to 480 mV for over 2 h from ambient humidity. Furthermore, a self-powered on-skin iontophoretic transdermal drug-delivery system is developed as an on-skin chemical intervention example. In addition, pencil–paper-based antennas, two-dimensional (2D) and three-dimensional (3D) circuits with light-emitting diodes (LEDs) and batteries, reconfigurable assembly and biodegradable electronics (based on water-soluble papers) are explored.

Significance: The rates of melanoma and nonmelanoma skin cancer are rising across the globe. Due to a shortage of board-certified dermatologists, the burden of dermal lesion screening and erythema monitoring has fallen to primary care physicians (PCPs). An adjunctive device for lesion screening and erythema monitoring would be beneficial because PCPs are not typically extensively trained in dermatological care. Aim: We aim to examine the feasibility of using a smartphone-camera-based dermascope and a USB-camera-based dermascope utilizing polarized white-light imaging (PWLI) and polarized multispectral imaging (PMSI) to map dermal chromophores and erythema. Approach: Two dermascopes integrating LED-based PWLI and PMSI with both a smartphone-based camera and a USB-connected camera were developed to capture images of dermal lesions and erythema. Image processing algorithms were implemented to provide chromophore concentrations and redness measures. Results: PWLI images were successfully converted to an alternate colorspace for erythema measures, and the spectral bandwidth of the PMSI LED illumination was sufficient for mapping of deoxyhemoglobin, oxyhemoglobin, and melanin chromophores. Both types of dermascopes were able to achieve similar relative concentration results. Conclusion: Chromophore mapping and erythema monitoring are feasible with PWLI and PMSI using LED illumination and smartphone-based cameras. These systems can provide a simpler, more portable geometry and reduce device costs compared with interference-filter-based or spectrometer-based clinical-grade systems. Future research should include a rigorous clinical trial to collect longitudinal data and a large enough dataset to train and implement a machine learning-based image classifier.

Significance: Research in tissue engineering and in vitro organ formation has recently intensified. To assess tissue morphology, the method of choice today is restricted primarily to histology. Thus novel tools are required to enable noninvasive, and preferably label-free, three-dimensional imaging that is more compatible with futuristic organ-on-a-chip models. Aim: We investigate the potential for using multiphoton microscopy (MPM) as a label-free in vitro approach to monitor calcium-induced epidermal differentiation. Approach: In vitro epidermis was cultured at the air–liquid interface in varying calcium concentrations. Morphology and tissue architecture were investigated using MPM based on visualizing cellular autofluorescence. Results: Distinct morphologies corresponding to epidermal differentiation were observed. In addition, Ca2+-induced effects could be distinguished based on the architectural differences in stratification in the tissue cultures. Conclusions: Our study shows that MPM based on cellular autofluorescence enables visualization of Ca2+-induced differentiation in epidermal skin models in vitro. The technique has potential to be further adapted as a noninvasive, label-free, and real-time tool to monitor tissue regeneration and organ formation in vitro.

Significance: Pulsatility is a vital characteristic of the cardiovascular system. Characterization of the pulsatility pattern locally in the peripheral microvasculature is currently not readily available and would provide an additional source of information which may prove important in understanding the pathophysiology of arterial stiffening, vascular ageing, and their linkage with cardiovascular disease development. Aim: We aim to confirm the suitability of speckle decorrelation optical coherence tomography angiography (OCTA) under various non-contact/contact scanning protocols for the visualization of pulsatility patterns in vessel-free tissue and in the microvasculature of peripheral human skin. Results: Results from 5 healthy subjects show distinct pulsatile patterns both in vessel-free tissue with either non-contact or contact imaging and in individual microvessels with contact imaging; respectively, likely caused by the pulsatile pressure and pulsatile blood flow. The pulse rates show good agreement with those from pulse oximetry, confirming that the pulsatile signatures reflect pulsatile hemodynamics. Conclusions: This study demonstrates the potential of speckle decorrelation OCTA for measuring localized peripheral cutaneous pulsatility and defines scanning protocols necessary to undertake such measurements. Non-contact imaging should be used for the study of pulsatility in vessel-free tissue and contact imaging with strong mechanical coupling in individual microvessels. Further studies of microcirculation based upon this method and protocols are warranted.

Significance: Spatial frequency domain imaging (SFDI) is a quantitative imaging method to measure absorption and scattering of tissue, from which several chromophore concentrations (e.g., oxy-/deoxy-/meth-hemoglobin, melanin, and carotenoids) can be calculated. Employing a method to extract additional spectral bands from RGB components (that we named cross-channels), we designed a handheld SFDI device to account for these pigments, using low-cost, consumer-grade components for its implementation and characterization. Aim: With only three broad spectral bands (red, green, blue, or RGB), consumer-grade devices are often too limited. We present a methodology to increase the number of spectral bands in SFDI devices that use RGB components without hardware modification. Approach: We developed a compact low-cost RGB spectral imager using a color CMOS camera and LED-based mini projector. The components’ spectral properties were characterized and additional cross-channel bands were calculated. An alternative characterization procedure was also developed that makes use of low-cost equipment, and its results were compared. The device performance was evaluated by measurements on tissue-simulating optical phantoms and in-vivo tissue. The measurements were compared with another quantitative spectroscopy method: spatial frequency domain spectroscopy (SFDS). Results: Out of six possible cross-channel bands, two were evaluated to be suitable for our application and were fully characterized (520 ± 20 nm; 556 ± 18 nm). The other four cross-channels presented a too low signal-to-noise ratio for this implementation. In estimating the optical properties of optical phantoms, the SFDI data have a strong linear correlation with the SFDS data (R2 = 0.987, RMSE = 0.006 for μa, R2 = 0.994, RMSE = 0.078 for μs′). Conclusions: We extracted two additional spectral bands from a commercial RGB system at no cost. There was good agreement between our device and the research-grade SFDS system. The alternative characterization procedure we have presented allowed us to measure the spectral features of the system with an accuracy comparable to standard laboratory equipment. Funding agencies: Knut and Alice Wallenberg FoundationKnut & Alice Wallenberg Foundation

Significance : Definitive diagnostics of many diseases is based on the histological analysis of thin tissue cuts with optical white light microscopy. Extra information on tissue structural properties obtained with polarized light would help the pathologist to improve the accuracy of his diagnosis. Aim: We report on using Mueller matrix microscopy data, logarithmic decomposition, and polarized Monte Carlo (MC) modeling for qualitative and quantitative analysis of thin tissue cuts to extract the information on tissue microstructure that is not available with a conventional white light microscopy. Approach: Unstained cuts of human skin equivalents were measured with a custom-built liquid-crystal-based Mueller microscope in transmission configuration. To interpret experimental data, we performed the simulations with a polarized MC algorithm for scattering anisotropic media. Several optical models of tissue (spherical scatterers within birefringent host medium, and combination of spherical and cylindrical scatterers within either isotropic or birefringent host medium) were tested. Results: A set of rotation invariants for the logarithmic decomposition of a Mueller matrix was derived to rule out the impact of sample orientation. These invariants were calculated for both simulated and measured Mueller matrices of the dermal layer of skin equivalents. We demonstrated that only the simulations with a model combining both spherical and cylindrical scatterers within birefringent host medium reproduced the experimental trends in optical properties of the dermal layer (linear retardance, linear dichroism, and anisotropic linear depolarization) with layer thickness. Conclusions: Our studies prove that Mueller polarimetry provides relevant information not only on a size of dominant scatterers (e.g., cell nuclei versus subwavelength organelles) but also on its shape (e.g., cells versus collagen fibers). The latter is directly related to the state of extracellular collagen matrix, which is often affected by early pathology. Hence, using polarimetric data can help to increase the accuracy of diagnosis.

We describe the contribution of our in vivo multiphoton microscopy (MPM) studies over the last ten years with DermaInspect® (JenLab, Germany), a CE-certified medical tomograph based on detection of fluorescent biomolecules, to the assessment of possible penetration of nanoparticulate zinc oxide in sunscreen through human skin. At the time we started our work, there was a strong movement for the precautionary principle to be applied to the use of nanoparticles in consumer products due to a lack of knowledge. The combined application of different MPM modalities, including spectral imaging, fluorescence lifetime imaging, second harmonic fluorescence generation, and phosphorescence microscopy, has provided overwhelming evidence that nanoparticle zinc oxide particles do not penetrate human skin when applied to various skin types with a range of methods of topical sunscreen application. MPM has also been used to study the viable epidermal morphology and redox state in supporting the safe use of topical zinc oxide nanoparticles. The impact of this work is emphasized by the recent proposed rule by the United States FDA on Sunscreen Drug Products for Over-the-Counter Human Use, which listed only zinc oxide and titanium dioxide of the currently marketed products to be generally recognized as safe and effective. Refereed/Peer-reviewed

Significance: Recent generation of bioengineered human skin allowed the efficient treatment of patients with severe skin defects. However, the optical and biomechanical properties of these models are not known. Aim: Three models of bioengineered human skin based on fibrin-agarose biomaterials (acellular, dermal skin substitutes, and complete dermoepidermal skin substitutes) were generated and analyzed. Approach: Optical and biomechanical properties of these artificial human skin substitutes were investigated using the inverse adding-doubling method and tensile tests, respectively. Results: The analysis of the optical properties revealed that the model that most resembled the optical behavior of the native human skin in terms of absorption and scattering properties was the dermoepidermal human skin substitutes after 7 to 14 days in culture. The time-course evaluation of the biomechanical parameters showed that the dermoepidermal substitutes displayed significant higher values than acellular and dermal skin substitutes for all parameters analyzed and did not differ from the control skin for traction deformation, stress, and strain at fracture break. Conclusions: We demonstrate the crucial role of the cells from a physical point of view, confirming that a bioengineered dermoepidermal human skin substitute based on fibrin-agarose biomaterials is able to fulfill the minimal requirements for skin transplants for future clinical use at early stages of in vitro development., Ministry of Science, Innovation and Universities of Spain PGC2018-101904-A-I0, Instituto de Salud Carlos III (ISCIII), Ministry of Science, Innovation and Universities, through AES 2017 AC17/00013, Instituto de Salud Carlos III (ISCIII), Ministry of Science, Innovation and Universities within EuroNanoMed framework, EU AC17/00013, University of Granada A.TEP.280.UGR18, Junta de Andalucía PE-0395-2019, Fundación Benéfica Anticancer San Francisco Javier y Santa Cándida, Granada, Spain, OTRI.35A-07

When analyzing multidiameter single-fiber reflectance (MDSFR) spectra, the inhomogeneous distribution of melanin pigments in skin tissue is usually not accounted for. Especially in heavily pigmented skins, this can result in bad fits and biased estimation of tissue optical properties. A model is introduced to account for the inhomogeneous distribution of melanin pigments in skin tissue. In vivo visible MDSFR measurements were performed on heavily pigmented skin of type IV to VI. Skin tissue optical properties and related physiological properties were extracted from the measured spectra using the introduced model. The absorption of melanin pigments described by the introduced model demonstrates a good correlation with the co-localized measurement of the well-known melanin index. (C) The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License.

Melanin is known to provide strong third-harmonic generation (THG) contrast in human skin. With a high concentration in basal cell cytoplasm, THG contrast provided by melanin overshadows other THG sources in human skin studies. For better understanding of the THG signals in keratinocytes without the influence of melanin, an in vivo THG microscopy (THGM) study was first conducted on vitiliginous skin. As a result, the THG-brightness ratio between the melanin-lacking cytoplasm of basal cells and collagen fibers is about 1.106 at the dermal–epidermal junctions of vitiliginous skin, indicating high sensitivity of THGM for the presence of melanin. We further applied the in vivo THGM to assist evaluating the therapeutic outcome from the histopathological point of view for those showed no improvement under narrowband ultraviolet B therapy based on the seven-point Physician Global Assessment score. Our clinical study indicates the high potential of THGM to assist the histopathological assessment of the therapeutic efficacy of vitiligo treatments.

Bisphenol A (BPA) is an endocrine disruptor frequently detected in human biofluids. Dermal absorption of BPA from thermal paper receipts occurs but BPA pharmacokinetics following dermal exposure is not understood. To compare the pharmacokinetics of dermal and dietary BPA exposure, six male participants handled simulated receipts containing relevant levels of BPA (isotope-labeled BPA-d16) for 5 min, followed by hand-washing 2 h later. Urine (0–48 h) and serum (0–7.5 h) were monitored for free and total BPA-d16. One week later, participants returned for a dietary administration with monitoring as above. One participant repeated the dermal administration with extended monitoring of urine (9 days) and serum (2 days). After dietary exposure, urine total BPA-d16 peaked within 5 h and quickly cleared within 24 h. After dermal exposure, cumulative excretion increased linearly for 2 days, and half the participants still had detectable urinary total BPA-d16 after 1 week. The participant repeating the dermal exposur...

A new sensitive non-invasive gold nanoparticle-based sensor that enables to detect thiols in the human skin has been developed. The detection procedure implied the assessment of the color change of a paper sensor resulting from aggregation of gold nanoparticles caused by thiols. The ratio of the intensity of the photo image blue channel vs the red one (in units of RGB coloration) served as analytical response. The main thiol in the skin is glutathione, therefore, it was used as model biothiol and spiking substance. The range of linearity for glutathione was 8–75 µM, the detection limit was 6.9 µM. RSD≤7% is for inter-day determination of 10 μM glutathione and RSD≤12% is the intra-day value. The recovery of 5 µM and 10 µM of glutathione was evaluated by applying solution, containing thiol-spikes, on skin. The results varied in the range 77–138%. A hundred-fold excess of serine, alanine, histidine, threonine, creatinine, urea, and ammonia; a ten-fold excess of glycine, proline, leucine, isoleucine, phenylalanine, asparagine; and a five-fold excess of valine, tryptophan, tyrosine, and uric acid, which can be extracted from the skin and is contained in the test matrix, have no significant effect on 10 µM glutathione signal. Thiols level in the skin of volunteers (21–65 years old, men and women) detected with the use of a proposed non-invasive sensor was 11.6–47.5 µM.

Non-contact vital sign monitoring enables the estimation of vital signs, such as heart rate, respiratory rate and oxygen saturation (SpO2), by measuring subtle color changes on the skin surface using a video camera. For patients in a hospital ward, the main challenges in the development of continuous and robust non-contact monitoring techniques are the identification of time periods and the segmentation of skin regions of interest (ROIs) from which vital signs can be estimated. We propose a deep learning framework to tackle these challenges. Approach: This paper presents two convolutional neural network (CNN) models. The first network was designed for detecting the presence of a patient and segmenting the patient’s skin area. The second network combined the output from the first network with optical flow for identifying time periods of clinical intervention so that these periods can be excluded from the estimation of vital signs. Both networks were trained using video recordings from a clinical study involving 15 pre-term infants conducted in the high dependency area of the neonatal intensive care unit (NICU) of the John Radcliffe Hospital in Oxford, UK. Main results: Our proposed methods achieved an accuracy of 98.8% for patient detection, a mean intersection-over-union (IOU) score of 88.6% for skin segmentation and an accuracy of 94.5% for clinical intervention detection using two-fold cross validation. Our deep learning models produced accurate results and were robust to different skin tones, changes in light conditions, pose variations and different clinical interventions by medical staff and family visitors. Significance: Our approach allows cardio-respiratory signals to be continuously derived from the patient’s skin during which the patient is present and no clinical intervention is undertaken.

pTwo-photon microscopes have been successfully translated into clinical imaging tools to obtain high-resolution optical biopsies foritalicin vivo/italichistology. We report on clinical multiphoton coherent anti-Stokes Raman spectroscopy (CARS) tomography based on two tunable ultrashort near-infrared laser beams for label-freeitalicin vivo/italicmultimodal skin imaging. The multiphoton biopsies were obtained with the compact tomograph "MPTflex-CARS" using a photonic crystal fiber, an optomechanical articulated arm, and a four-detector-360 deg measurement head. The multiphoton tomograph has been employed to patients in a hospital with diseased skin. The clinical study involved 16 subjects, 8 patients with atopic dermatitis, 4 patients with psoriasis vulgaris, and 4 volunteers served as control. Two-photon cellular autofluorescence lifetime, second harmonic generation (SHG) of collagen, and CARS of intratissue lipids/proteins have been detected with single-photon sensitivity, submicron spatial resolution, and picosecond temporal resolution. The most important signal was the autofluorescence from nicotinamide adenine dinucleotide [NAD(P)H]. The SHG signal from collagen was mainly used to detect the epidermal-dermal junction and to calculate the ratio elastin/collagen. The CARS/Raman signal provided add-on information. Based on this view on the disease-affected skin on a subcellular level, skin areas affected by dermatitis and by psoriasis could be clearly identified. Multimodal multiphoton tomographs may become important label-free clinical high-resolution imaging tools foritalicin vivo/italicskin histology to realize rapid early diagnosis as well as treatment control./p.

Photoacoustic microscopy (PAM) is a fast-growing biomedical imaging technique that provides high-resolution in vivo imaging beyond the optical diffusion limit. Depending on the scalable lateral resolution and achievable penetration depth, PAM can be classified into optical resolution PAM (OR-PAM) and acoustic resolution PAM (AR-PAM). The use of a microelectromechanical systems (MEMS) scanner has improved OR-PAM imaging speed significantly and is highly beneficial in the development of miniaturized handheld devices. The shallow penetration depth of OR-PAM limits the use of such devices for a wide range of clinical applications. We report the use of a high-speed MEMS scanner for both OR-PAM and AR-PAM. A high-speed, wide-area scanning integrated OR-AR-PAM system combining MEMS scanner and raster mechanical movement was developed. A lateral resolution of 5 μm and penetration depth ∼0.9-mm in vivo was achieved using OR-PAM at 586 nm, whereas a lateral resolution of 84 μm and penetration depth of ∼2-mm in vivo was achieved using AR-PAM at 532 nm.

The human microbiome encompasses the fungi, bacteria and viruses that live on, within, and immediately surrounding the body. Microbiomes have potential utility in forensic science as an evidentiary tool to link or exclude persons of interest associated with criminal activities. Research has shown the microbiome is individualised, and that personal microbial signatures can be recovered from surfaces such as phones, shoes and fabrics. Before the human microbiome may be used as an investigative tool, further research is required to investigate the utility and potential limitations surrounding microbial profiling. This includes the detectability of microbial transfer between individuals or items, the associated risks (such as contamination events) and the applicability of microbial profiling for forensic purposes. This research aimed to identify whether an individual's distinguishable microbiome could be transferred to another individual and onto substrates, and vice versa. Paper, cotton, and glass surfaces were chosen to represent a range of substrate matrices. The study involved six participants placed into three pairs; participants took part in two modes of transfer. Transfer Mode 1 involved the pair shaking hands, followed by rubbing a substrate in their right hand. Transfer Mode 2 involved individuals rubbing a substrate in their left hand, swapping substrates with their partner and then rubbing the swapped substrate in their left hand. 16S rRNA sequencing was performed on the extracted microbial DNA from participant and substrate samples. Quantitative Insights into Microbial Ecology 2 (QIIME 2) was used for sequence quality control and beta (between-sample) diversity analyses and taxonomic assignment. Principal Coordinate Analysis (PCoA) based on Jaccard distances was visualised through Emperor software to determine the phylogenetic similarity of bacterial communities between participants and among participant pairs. Statistical testing through PERMANOVA revealed significant differences in the Jaccard distances between each participant pair (P 0.001), highlighting not only the potential distinguishability of skin microbiomes among individuals, but also the clustering effect observed between participant pairs due to the potential transfer of hand-associated microbiomes between individuals. The study demonstrated that transfer of the human skin microbiome had occurred between all participant pairs, regardless of substrate type or mode of transfer.

The silicon photomultiplier (SIPM) is an emerging detector technology that enables both high sensitivity and high dynamic range detection of visible and near-infrared light at a fraction of the cost of conventional vacuum tube photomultiplier tubes (PMTs). A low-cost detection circuit is presented and the performance of a commercial SIPM is evaluated for high-speed laser scanning microscopy applications. For moderate-to-high-speed fluorescent imaging applications, the measurements and imaging results indicate that the SIPM exceeds the sensitivity of GaAsP PMTs, while providing higher dynamic range and better saturation behavior. For low speed or applications requiring large detector areas, the GaAsP PMT retains a sensitivity advantage due to large area and lower dark counts. The calculations presented show that, above a critical detection bandwidth, the SIPM sensitivity exceeds that of a GaAsP PMT.