Hyperspectral imaging to accurately segment skin erythema and hyperpigmentation in cutaneous chronic graft-versus-host disease.

In 51 lesions from 15 patients with the inflammatory skin condition chronic graft-versus-host-disease, hyperspectral imaging accurately delineated active erythema and post-inflammatory hyperpigmentation. The method was validated by dermatologist-approved confident delineations of only definitely affected and definitely unaffected areas in photographs. A prototype hyperspectral imaging system acquired a 2.5 × 3.5 cm2 area of skin at 120 wavelengths in the 450-850 nm range. Unsupervised extraction of unknown absorbers by endmember analysis achieved a comparable accuracy to that of supervised extraction of known absorbers (melanin, hemoglobin) by chromophore mapping: 0.78 (IQR: 0.39-0.85) vs. 0.83 (0.53-0.91) to delineate erythema and 0.74 (0.57-0.87) vs. 0.73 (0.52-0.84) to delineate hyperpigmentation. Both algorithms achieved higher specificity than sensitivity. Whereas a trained human confidently marked a median of 7% of image pixels, unsupervised and supervised algorithms delineated a median of 14% and 27% pixels. Hyperspectral imaging could overcome a fundamental practice gap of distinguishing active from inactive manifestations of inflammatory skin disease.

Three-dimensional representation of triple spectral line imaging data as an option for noncontact skin diagnostics.

SIGNIFICANCE: Skin malformations in dermatology are mostly evaluated subjectively, based on a doctor's experience and visual perception; an option for objective quantitative skin assessment is camera-based spectrally selective diagnostics. Multispectral imaging is a technique capable to provide information about concentrations of the absorbing chromophores and their distribution over the malformation in a noncontact way. Conversion of spectral images into distribution maps of chromophores can be performed by means of the modified Beer-Lambert law. However, such distribution maps represent only single specific cases, therefore, some extensive method for data comparison is needed. AIM: This study aims to develop a more informative approach for identification and characterization of skin malformations using three-dimensional (3D) representation of triple spectral line imaging data. APPROACH: The 3D-representation method is experimentally tested on eight different skin pathology types, including both benign and malignant pathologies; an imaging device ensuring uniform three laser line (448, 532, and 659 nm) illumination is used. Three spectral line images are extracted from a single snapshot RGB image data, with subsequent calculation of attenuation coefficients for each working wavelength at every image pixel and represented as 3D graphs. Skin chromophore content variations in malformations are represented in a similar way. RESULTS: Clinical measurement results for 99 skin pathologies, including basal cell carcinomas, melanoma, dermal nevi, combined nevi, junctional nevi, blue nevi, seborrheic keratosis, and hemangiomas. They are presented as 3D spectral attenuation maps exhibiting specific individual features for each group of pathologies. Along with intensity attenuation maps, 3D maps for content variations of three main skin chromophores (melanin, oxyhemoglobin, and deoxyhemoglobin), calculated in frame of a model based on modified Beer-Lambert law, are also presented. Advantages and disadvantages of the proposed data representation method are discussed. CONCLUSIONS: The described 3D-representation method of triple spectral line imaging data shows promising potential for objective quantitative noncontact diagnosis of skin pathologies.

Profiling and imaging of forensic evidence - A pan-European forensic round robin study part 1: Document forgery.

The forensic scenario, on which the round robin study was based, simulated a suspected intentional manipulation of a real estate rental agreement consisting of a total of three pages. The aims of this study were to (i) establish the amount and reliability of information extractable from a single type of evidence and to (ii) provide suggestions on the most suitable combination of compatible techniques for a multi-modal imaging approach to forgery detection. To address these aims, seventeen laboratories from sixteen countries were invited to answer the following tasks questions: (i) which printing technique was used? (ii) were the three pages printed with the same printer? (iii) were the three pages made from the same paper? (iv) were the three pages originally stapled? (v) were the headings and signatures written with the same ink? and (vi) were headings and signatures of the same age on all pages? The methods used were classified into the following categories: Optical spectroscopy, including multispectral imaging, smartphone mapping, UV-luminescence and LIBS; Infrared spectroscopy, including Raman and FTIR (micro-)spectroscopy; X-ray spectroscopy, including SEM-EDX, PIXE and XPS; Mass spectrometry, including ICPMS, SIMS, MALDI and LDIMS; Electrostatic imaging, as well as non-imaging methods, such as non-multimodal visual inspection, (micro-)spectroscopy, physical testing and thin layer chromatography. The performance of the techniques was evaluated as the proportion of discriminated sample pairs to all possible sample pairs. For the undiscriminated sample pairs, a distinction was made between undecidability and false positive claims. It was found that none of the methods used were able to solve all tasks completely and/or correctly and that certain methods were a priori judged unsuitable by the laboratories for some tasks. Correct results were generally achieved for the discrimination of printer toners, whereas incorrect results in the discrimination of inks. For the discrimination of paper, solid state analytical methods proved to be superior to mass spectrometric methods. None of the participating laboratories deemed addressing ink age feasible. It was concluded that correct forensic statements can only be achieved by the complementary application of different methods and that the classical approach of round robin studies to send standardised subsamples to the participants is not feasible for a true multimodal approach if the techniques are not available at one location.

Skin chromophore mapping by smartphone RGB camera under spectral band and spectral line illumination.

SIGNIFICANCE: Multispectral imaging enables mapping of chromophore content changes in skin neoplasms, which helps to diagnose a pathology. Different types of light sources can be used for the imaging. Design of laser-based illuminators is more complicated and, consequently, they are more expensive than LED-based illuminators. On the other hand, spectral line illumination has the advantage of less complicated calculations, since only the discrete maximum wavelengths need to be considered. Spectral band and spectral line approaches for multispectral skin diagnostics have not been compared so far. This can help to evaluate the accuracy and effectiveness of both approaches. AIM: To compare two specific illumination modalities-spectral band and spectral line illumination-from the point of performance for mapping of in vivo skin chromophores. APPROACH: Three spectral images of the same skin malformations were captured by a smartphone RGB camera with two different add-on illuminators comprising LED emitters and laser emitters, respectively. Five types of benign skin neoplasms were included in our study. Concentrations of skin melanin, oxy- and deoxy-hemoglobin at image pixel groups were calculated using the Beer-Lambert law. RESULTS: Skin chromophore maps and statistical analysis of mean concentrations' changes in the neoplasms compared to the surrounding skin are presented and discussed. The data of the laser emitters led to significantly higher (∼10 times) increase of the oxy-hemoglobin values in vascular neoplasms and much lower deoxy-hemoglobin values, if compared to the data obtained by the LED emitters. CONCLUSIONS: Analysis of the obtained chromophore distribution maps and concentration variations in neoplasms led to conclusion that the spectral line illumination approach is more appropriate for this application. Considering only the peak wavelengths of illumination spectral bands leads to essentially different results if compared to those obtained by spectral line illumination and may cause misinterpretations in the clinical assessment of skin neoplasms.

Beer-Lambert law for optical tissue diagnostics: current state of the art and the main limitations.

SIGNIFICANCE: Beer-Lambert law (BLL) is a widely used tool for contact and remote determination of absorber concentration in various media, including living tissues. Originally proposed in the 18th century as a simple exponential expression, it has survived numerous modifications and updates. The basic assumptions of this law may not be fulfilled in real measurement conditions. This can lead to mistaken or misinterpreted results. In particular, the effects to be additionally taken into account in the tissue measurements include anisotropy, scattering, fluorescence, chemical equilibria, interference, dichroism, spectral bandwidth disagreements, stray radiation, and instrumental effects. AIM: We review the current state of the art and the main limitations of remote tissue diagnostics using the BLL. Historical development of updating this law by taking into account specific additional factors such as light scattering and photon pathlengths in diffuse reflectance is described, along with highlighting the main risks to be considered by interpreting the measured data. APPROACH: Literature data related to extension and modification of the BLL related to tissue assessment and concentration estimation of specific tissue molecules are collected and analyzed. The main emphasis here is put on the optical measurements of living tissue chromophore concentrations and estimation of physiological parameters, e.g., blood oxygen saturation. RESULTS: Modified expressions of the BLL suitable for several specific cases of living tissue characterization are presented and discussed. CONCLUSIONS: Applications of updated/modified Beer-Lambert law (MBLL) with respect to particular measurement conditions are helpful for obtaining more reliable data on the target tissue physiological state and biochemical content. MBLL accounting for the role of scattering in several ways appears to be a successful approach. Extended MBLL and BLL in the time domain form could provide more accurate results, but this requires more time resources to be spent.

Autofluorescence imaging for recurrence detection in skin cancer postoperative scars.

This clinical study is a first attempt to use autofluorescence for recurrence diagnosis of skin cancer in postoperative scars. The proposed diagnostic parameter is based on a reduction in scar autofluorescence, evaluated in the green spectral channel. The validity of the method has been tested on 110 postoperative scars from 56 patients suspected of non-melanoma skin cancer, with eight patients (13 scars) available for the repeated examination. The recurrence diagnosis within a scar has been made after two subsequent autofluorescence check-ups, representing the temporal difference between the scar autofluorescence amplitudes as a vector. The recognition of recurrence has been discussed to represent the significant deviations from the value of vector angle θ. This new autofluorescence-based method can be easily integrated into the postoperative monitoring of surgical scars and can help diagnose the recurrence of skin cancer from the early stage of scar development.

Photoplethysmography in dogs and cats: a selection of alternative measurement sites for a pet monitor.

OBJECTIVE: Photoplethysmography (PPG) is an increasingly popular health and well-being tool for monitoring heart rate and oxygen saturation. Due to the pigmentation and hairiness of dogs and cats, a pulse oximeter is routinely placed solely on the tongue. As this approach is feasible only for pet monitor use during surgical procedures, we investigate PPG signal quality on several other measurement sites that would be better tolerated by conscious animals. APPROACH: Acquired PPG signals are analyzed by four signal quality indices: mean baseline, signal power, kurtosis, and tolerance score. MAIN RESULTS: In dogs, the metacarpus and tail can be substituted for oral pulse oximeter placement since both measurement sites exhibited high PPG signal kurtosis and were considered well-tolerated. In cats, the digit could be used with some limitations. SIGNIFICANCE: Pet monitors with pulse oximeter probes adjusted to promising measurement sites could enable veterinarians and owners to monitor animals when fully awake.

Differentiation of seborrheic keratosis from basal cell carcinoma, nevi and melanoma by RGB autofluorescence imaging.

A clinical trial on the autofluorescence imaging of skin lesions comprising 16 dermatologically confirmed pigmented nevi, 15 seborrheic keratosis, 2 dysplastic nevi, histologically confirmed 17 basal cell carcinomas and 1 melanoma was performed. The autofluorescence spatial properties of the skin lesions were acquired by smartphone RGB camera under 405 nm LED excitation. The diagnostic criterion is based on the calculation of the mean autofluorescence intensity of the examined lesion in the spectral range of 515 nm-700 nm. The proposed methodology is able to differentiate seborrheic keratosis from basal cell carcinoma, pigmented nevi and melanoma. The sensitivity and specificity of the proposed method was estimated as being close to 100%. The proposed methodology and potential clinical applications are discussed in this article.

Multispectral, Fluorescent and Photoplethysmographic Imaging for Remote Skin Assessment.

Optical tissue imaging has several advantages over the routine clinical imaging methods, including non-invasiveness (it does not change the structure of tissues), remote operation (it avoids infections) and the ability to quantify the tissue condition by means of specific image parameters. Dermatologists and other skin experts need compact (preferably pocket-size), self-sustaining and easy-to-use imaging devices. The operational principles and designs of ten portable in-vivo skin imaging prototypes developed at the Biophotonics Laboratory of Institute of Atomic Physics and Spectroscopy, University of Latvia during the recent five years are presented in this paper. Four groups of imaging devices are considered. Multi-spectral imagers offer possibilities for distant mapping of specific skin parameters, thus facilitating better diagnostics of skin malformations. Autofluorescence intensity and photobleaching rate imagers show a promising potential for skin tumor identification and margin delineation. Photoplethysmography video-imagers ensure remote detection of cutaneous blood pulsations and can provide real-time information on cardiovascular parameters and anesthesia efficiency. Multimodal skin imagers perform several of the abovementioned functions by taking a number of spectral and video images with the same image sensor. Design details of the developed prototypes and results of clinical tests illustrating their functionality are presented and discussed.

Smartphone snapshot mapping of skin chromophores under triple-wavelength laser illumination.

Chromophore distribution maps are useful tools for skin malformation severity assessment and for monitoring of skin recovery after burns, surgeries, and other interactions. The chromophore maps can be obtained by processing several spectral images of skin, e.g., captured by hyperspectral or multispectral cameras during seconds or even minutes. To avoid motion artifacts and simplify the procedure, a single-snapshot technique for mapping melanin, oxyhemoglobin, and deoxyhemoglobin of in-vivo skin by a smartphone under simultaneous three-wavelength (448­532­659 nm) laser illumination is proposed and examined. Three monochromatic spectral images related to the illumination wavelengths were extracted from the smartphone camera RGB image data set with respect to crosstalk between the RGB detection bands. Spectral images were further processed accordingly to Beer's law in a three chromophore approximation. Photon absorption path lengths in skin at the exploited wavelengths were estimated by means of Monte Carlo simulations. The technique was validated clinically on three kinds of skin lesions: nevi, hemangiomas, and seborrheic keratosis. Design of the developed add-on laser illumination system, image-processing details, and the results of clinical measurements are presented and discussed.

Autofluorescence imaging of basal cell carcinoma by smartphone RGB camera.

The feasibility of smartphones for in vivo skin autofluorescence imaging has been investigated. Filtered autofluorescence images from the same tissue area were periodically captured by a smartphone RGB camera with subsequent detection of fluorescence intensity decreasing at each image pixel for further imaging the planar distribution of those values. The proposed methodology was tested clinically with 13 basal cell carcinoma and 1 atypical nevus. Several clinical cases and potential future applications of the smartphone-based technique are discussed.

Study of smartphone suitability for mapping of skin chromophores.

RGB (red-green-blue) technique for mapping skin chromophores by smartphones is proposed and studied. Three smartphones of different manufacturers were tested on skin phantoms and in vivo on benign skin lesions using a specially designed light source for illumination. Hemoglobin and melanin indices obtained by these smartphones showed differences in both tests. In vitro tests showed an increment of hemoglobin and melanin indices with the concentration of chromophores in phantoms. In vivo tests indicated higher hemoglobin index in hemangiomas than in nevi and healthy skin, and nevi showed higher melanin index compared to the healthy skin. Smartphones that allow switching off the automatic camera settings provided useful data, while those with "embedded" automatic settings appear to be useless for distant skin chromophore mapping.

Snapshot RGB mapping of skin melanin and hemoglobin.

The concept of snapshot red-green-blue (RGB) multispectral imaging was applied for skin chromophore mapping. Three monochromatic spectral images have been extracted from a single RGB image dataset at simultaneous illumination of skin by 473-, 532-, and 659-nm laser lines. The spectral images were further transformed into distribution maps of skin melanin, oxyhemoglobin, and deoxyhemoglobin, related to pigmented and vascular skin malformations. The performance and clinical potential of the proposed technique are discussed

Photobleaching effects on in vivo skin autofluorescence lifetime.

The autofluorescence lifetime of healthy human skin was measured using excitation provided by a picosecond diode laser operating at a wavelength of 405 nm and with fluorescence emission collected at 475 and 560 nm. In addition, spectral and temporal responses of healthy human skin and intradermal nevus in the spectral range 460 to 610 nm were studied before and after photobleaching. A decrease in the autofluorescences lifetimes changes was observed after photobleaching of human skin. A three-exponential model was used to fit the signals, and under this model, the most significant photoinduced changes were observed for the slowest lifetime component in healthy skin at the spectral range 520 to 610 nm and intradermal nevus at the spectral range 460 to 610 nm.

Noncontact monitoring of vascular lesion phototherapy efficiency by RGB multispectral imaging.

A prototype low-cost RGB imaging system consisting of a commercial RGB CMOS sensor, RGB light-emitting diode ring light illuminator, and a set of polarizers was designed and tested for mapping the skin erythema index, in order to monitor skin recovery after phototherapy of vascular lesions, such as hemangiomas and telangiectasias. The contrast of erythema index (CEI) was proposed as a parameter for quantitative characterization of vascular lesions. Skin recovery was characterized as a decrease of the CEI value relative to the value before the treatment. This approach was clinically validated by examining 31 vascular lesions before and after phototherapy.

Clinical evaluation of melanomas and common nevi by spectral imaging.

A clinical trial on multi-spectral imaging of malignant and non-malignant skin pathologies comprising 17 melanomas and 65 pigmented common nevi was performed. Optical density data of skin pathologies were obtained in the spectral range 450-950 nm using the multispectral camera Nuance EX. An image parameter and maps capable of distinguishing melanoma from pigmented nevi were proposed. The diagnostic criterion is based on skin optical density differences at three fixed wavelengths: 540nm, 650nm and 950nm. The sensitivity and specificity of this method were estimated to be 94% and 89%, respectively. The proposed methodology and potential clinical applications are discussed.

Towards noncontact skin melanoma selection by multispectral imaging analysis.

A clinical trial comprising 334 pigmented and vascular lesions has been performed in three Riga clinics by means of multispectral imaging analysis. The imaging system Nuance 2.4 (CRi) and self-developed software for mapping of the main skin chromophores were used. Specific features were observed and analyzed for malignant skin melanomas: notably higher absorbance (especially as the difference of optical density relative to the healthy skin), uneven chromophore distribution over the lesion area, and the possibility to select the "melanoma areas" in the correlation graphs of chromophores. The obtained results indicate clinical potential of this technology for noncontact selection of melanoma from other pigmented and vascular skin lesions.

Contact and contactless diffuse reflectance spectroscopy: potential for recovery monitoring of vascular lesions after intense pulsed light treatment.

Optical fiber contact probe diffuse reflectance spectroscopy and remote multispectral imaging methods in the spectral range of 400 to 1100 nm were used for skin vascular malformation assessment and recovery tracing after treatment by intense pulsed light. The results confirmed that oxy-hemoglobin relative changes and the optical density difference between lesion and healthy skin in the spectral region 500 to 600 nm may be successfully used for objective appraisal of the therapy effect. Color redness parameter a* = 2 is suggested as a diagnostic border to distinguish healthy skin and vascular lesions, and as the indicator of phototreatment efficiency. Valuable diagnostic information on large area (>5 mm) lesions and lesions with uncertain borders can be proved by the multispectral imaging method.

2-D mapping of skin chromophores in the spectral range 500 - 700 nm.

The multi-spectral imaging technique has been used for distant mapping of in-vivo skin chromophores by analyzing spectral data at each reflected image pixel and constructing 2-D maps of the relative concentrations of oxy-/deoxy-haemoglobin and melanin. Instead of using a broad visible-NIR spectral range, this study focuses on narrowed spectral band 500-700 nm, speeding-up the signal processing procedure. Regression analysis confirmed that superposition of three Gaussians is optimal analytic approximation for the oxy-haemoglobin absorption tabular spectrum in this spectral band, while superposition of two Gaussians fits well for deoxy-haemoglobin absorption and exponential function - for melanin absorption. The proposed approach was clinically tested for three types of in-vivo skin provocations: ultraviolet irradiance, chemical reaction with vinegar essence and finger arterial occlusion. Spectral range 500-700 nm provided better sensitivity to oxy-haemoglobin changes and higher response stability to melanin than two reduced ranges 500-600 nm and 530-620 nm.

Imaging of laser-excited tissue autofluorescence bleaching rates.

Experimental methodology for imaging of laser-excited tissue autofluorescence bleaching rates has been developed and clinically tested. The fluorescence images were periodically captured from the same tissue area over a certain time, with subsequent detection of the fluorescence intensity decrease rate at each image pixel and further imaging the planar distribution of those values. Spectral features at each image pixel were analyzed with a hyperspectral imaging camera. Details of the equipment and image processing are described as well as some measurement results that confirm the feasibility of the proposed technology.