Evaluation of Skin Vascular Malformations’ Laser Treatment by RGB and Multi-spectral Imaging
RGB imaging system for mapping and monitoring of hemoglobin changes in skin has been tested for evaluation of vascular malformations’ laser treatment. The multi-spectral imaging system was used as the reference.
Application of principal component analysis to multispectral imaging data for evaluation of pigmented skin lesions
Non-invasive and fast primary diagnostics of pigmented skin lesions is required due to frequent incidence of skin cancer – melanoma. Diagnostic potential of principal component analysis (PCA) for distant skin melanoma recognition is discussed. Processing of the measured clinical multi-spectral images (31 melanomas and 94 nonmalignant pigmented lesions) in the wavelength range of 450-950 nm by means of PCA resulted in 87 % sensitivity and 78 % specificity for separation between malignant melanomas and pigmented nevi.
LASCA and PPG imaging for non-contact assessment of skin blood supply
Laser speckle contrast analysis (LASCA) offers a non-contact, full-field, and real-time mapping of capillary blood flow and can be considered as an alternative method to Laser Doppler perfusion imaging (LDPI). Photoplethysmography (PPG) is well known technique for assessment of skin blood pulsations that can be related to blood flow. In recent years several studies have been done on development of non-contact PPG imaging (PPGI). LASCA and PPGI techniques are simpler and cheaper compared with LDPI. LASCA technique has been implemented in several commercial instruments. However, these systems are still too expensive and bulky to be widely available. Several optical techniques have found new i…
Development of a non-invasive LED based device for adipose tissue thickness measurements in vivo
There are a number of techniques for body composition assessment in clinics and in field-surveys, but in all cases the applied methods have advantages and disadvantages. High precision imaging methods are available, though expensive and non-portable, however, the methods devised for the mass population, often suffer from the lack of precision. Therefore, the development of a safe, mobile, non-invasive, optical method that would be easy to perform, precise and low-cost, but also would offer an accurate assessment of subcutaneous adipose tissue (SAT) both in lean and in obese persons is required. Thereof, the diffuse optical spectroscopy is advantageous over the aforementioned techniques. A p…
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. Spe- cific 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 correla- tion graphs of chromophores. The obtained results indicate clinical potential of this technology for noncontact selec- tion of mel…
Benign — A typical nevi discrimination using diffuse reflectance and fluorescence multispectral imaging system
Early diagnostics of skin cancer is of interest for dermatologists. Atypical nevi are not considered to be malignant, but are suspects that should be detected and monitored over time. The multispectral imaging system Nuance operating in spectral range 450–950 nm was adapted for clinical in vivo measurements in diffuse reflectance and fluorescence mode. Mean and standard deviation values of optical density and fluorescence intensity were extracted from segmented pigmented lesions (21 benign and 26 atypical nevi) and used for further analysis. It was possible to achieve 62% sensitivity and 67% specificity for discrimination between atypical and benign lesions using averaged fluorescence mean …
Implementation of laser speckle contrast analysis as connection kit for mobile phone for assessment of skin blood flow
Laser speckle contrast analysis (LASCA) offers a non-contact, full-field, and real-time mapping of capillary blood flow and can be considered as an alternative method to Laser Doppler perfusion imaging. LASCA technique has been implemented in several commercial instruments. However, these systems are still too expensive and bulky to be widely available. Several optical techniques have found new implementations as connection kits for mobile phones thus offering low cost screening devices. In this work we demonstrate simple implementation of LASCA imaging technique as connection kit for mobile phone for primary low-cost assessment of skin blood flow. Stabilized 650 nm and 532 nm laser diode m…
Diffuse reflectance and fluorescence multispectral imaging system for assessment of skin
The diffuse reflectance multispectral imaging technique has been used for distant mapping of in vivo skin chromophores (hemoglobin and melanin). The fluorescence multispectral imaging is not so common for skin applications due to complicity of data acquisition and processing, but could provide additional information about skin fluorophores. Both techniques are compatible, and could be combined into a multimodal solution. The multispectral imaging system Nuance based on liquid crystal tunable filters was adapted for diffuse reflectance and fluorescence spectral imaging of in vivo skin. Uniform illumination was achieved by LED ring light. Combination of four LEDs (warm white, 770 nm, 830 nm a…
Comparison of single-spot technique and RGB imaging for erythema index estimation
A commercially available point measurement device, the Mexameter(®), and an experimental RGB imaging prototype device were used for erythema index estimation of 50 rosacea patients by analysing the level of skin redness on the forehead, both cheeks and both sides of a nose. Results are compared with Clinician's Erythema Assessment (CEA) values given by two dermatologists. The Mexameter uses 568 nm and 660 nm LEDs and a photodetector for estimation of erythema index, while the used prototype device acquired RGB images at 460 nm, 530 nm and 665 nm LED illumination. Several erythema index estimation algorithms were compared to determine which one gives the best contrast between increased eryth…
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 mel…
Multi-spectral imaging analysis of pigmented and vascular skin lesions: results of a clinical trial
A clinical trial comprising 266 pigmented lesions and 49 vascular lesions has been performed in three Riga clinics by means of multi-spectral imaging analysis. The imaging system Nuance 2.4 (CRI) and self-developed software for mapping of the main skin chromophores were used. The obtained results confirm clinical potential of this technology for non-contact quantitative assessment of skin pathologies.
RGB imaging of laser-excited skin autofluorescence bleaching rates
In-vivo skin photo-bleaching (intensity decrease during irradiation) has been investigated at 405 nm cw laser excitation. Digital RGB photo-camera was used for studies of the bleaching features by analysis of fluorescent images at separated R, G and B spectral bands. Diagnostic potential based on correlations between skin pigmentation and bleaching rates is discussed.
Influence of low power CW laser irradiation on skin hemoglobin changes
Influence of low power laser irradiance on healthy skin using diffuse reflectance spectroscopy and multispectral imaging was studied. Changes of diffuse reflectance spectra in spectral range from 500 to 600 nm were observed after 405 nm, 473 nm and 532 nm laser provocation, leading to conclusion that the content of skin hemoglobin has changed. Peaks in spectral absorbance (optical density) curves corresponded to well-known oxy-hemoglobin absorbance peaks at 542 and 577 nm.
Mobile phone based laser speckle contrast imager for assessment of skin blood flow
Assessment of skin blood flow is of interest for evaluation of skin viability as well as for reflection of the overall condition of the circulatory system. Laser Doppler perfusion imaging (LDPI) and laser speckle contrast imaging (LASCI) are optical techniques used for assessment of skin perfusion. However, these systems are still too expensive and bulky to be widely available. Implementation of such techniques as connection kits for mobile phones have a potential for primary diagnostics. In this work we demonstrate simple and low cost LASCI connection kit for mobile phone and its comparison to laser Doppler perfusion imager. Post-occlusive hyperemia and local thermal hyperemia tests are us…
Mobile platform for online processing of multimodal skin optical images: Using online Matlab server for processing remission, fluorescence and laser speckle images, obtained by using novel handheld device
Mobile platform for multimodal skin assessment has been developed. Different illumination sources allow switching between modalities. Diffuse reflectance spectral imaging is provided by five LEDs, fluorescence is excited by 405 nm LEDs, and laser speckle by 650 nm laser diode. Handheld, battery powered device includes all light sources and color camera with USB. The core of the system is Linux OS embedded microcontroller. USB, FTP and HTML with JavaScript combination is used to create standard image transfer and control interface. In combination with built in WiFi access point it allows online skin images storage and processing. It means that data processing algorithms are located and updat…
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.
Real-Time Photoplethysmography Imaging System
Real-time non-contact photoplethysmography imaging (PPGI) system for high-resolution blood perfusion mapping in human skin has been proposed. The PPGI system comprises of LED lamp, webcam and computer with video processing software. The purpose of this study is to evaluate the reliability of the PPGI system when measuring blood perfusion. The validation study of PPGI and laser-Doppler perfusion imager (LDPI) was performed during local warming of palm skin. Results showed that the amplitude of PPGI increases immediately after warming and well correlated with the mean LDPI amplitude (R=0.92+-0.03, p<0.0001). We found that PPGI technique has good potential for non-contact monitoring of blood p…
RGB imaging system for monitoring of skin vascular malformation's laser therapy
A prototype RGB imaging system for mapping of skin chromophores consists of a commercial RGB CMOS sensor, RGB LEDs ring-light illuminator and orthogonally orientated polarizers for reducing specular reflectance. The system was used for monitoring of vascular malformations (hemagiomas and telangiectasias) therapy.
Melanoma-Nevus Discrimination Based on Image Statistics in Few Spectral Channels
The purpose of this paper is to offer a method for discrimination of cutaneous melanoma from benign nevus, founded on analysis of skin lesion image. At the core of method is calculation of mean and standard deviation of pixel optical density values for a few narrow spectral bands. Calculated values are compared with discriminating thresholds derived from a set of images of benign nevi and melanomas with known diagnosis. Classification is done applying weighted majority rule to results of thresholding. Verification against the available multispectral images of 32 melanomas and 94 benign nevi has shown that the method using three spectral bands provided zero false negative and four false posi…
Skin haemoglobin mapping: comparison of multi-spectral imaging and selective R-G-B analysis
The multi-spectral imaging technique has been used for distant mapping of in-vivo skin haemoglobin. Besides, potential of selective R-G-B analysis of skin images has been studied under bi-chromatic (532 nm and 635 nm) laser illumination.
RGB mapping of hemoglobin distribution in skin
An experimental RGB imaging system based on commercial color camera was constructed, and its potential for mapping of hemoglobin distribution in skin was studied. Two types of LEDs (RGB and white “warm” LEDs) were compared as illuminators for acquiring images of vascular and pigmented skin malformations. A novel approach for studies of skin capillary refill by RGB analysis has been proposed and discussed.
Multi-spectral mapping of in vivo skin hemoglobin and melanin
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-/deoxyhemoglobin and melanin. Instead of using a broad visible-NIR spectral range, this study focuses on narrowed spectral band 500-700 nm, so speeding-up the signal processing procedure. Regression analysis confirmed that superposition of three Gaussians is optimal analytic approximation for the oxy-hemoglobin absorption tabular spectrum in this spectral band, while superposition of two Gaussians fits well for deoxy-hemoglobin absorption and exponential function - for mela…
RGB imaging system for mapping and monitoring of hemoglobin distribution in skin
A prototype R-G-B imaging system for mapping of skin hemoglobin distribution has been designed and tested. Device basically consists of a commercial RGB sensor (CMOS, max. frame rate 87 fps for VGA resolution), RGB LED ringlight illuminator and orthogonally orientated polarizers for reducing specular reflectance. The system was examined for monitoring of hemoglobin concentration changes during specific provocations - arterial/venous occlusions and heat test. Hemoglobin distribution maps of several skin malformations were obtained, as well.
Assessment of efficiencies of electroporation and sonoporation methods by using fluorescence RGB imaging method
Simple RGB method for fluorescence in vivo imaging is presented to assess efficiency of electroporation and sonoporation methods by measuring distribution and accumulation of green fluorescence protein (GFP) concentration. 20 laboratory measurements were performed on mice to test the method.
Determination of chromophore distribution in skin by spectral imaging
Possibilities to determine chromophore distribution in skin by spectral imaging were explored. Simple RGB sensor devices were used for image acquisition. Totally 200 images of 40 different bruises of 20 people were obtained in order to map chromophores bilirubin and haemoglobin. Possibilities to detect water in vitro and in vivo were estimated by using silicon photodetectors and narrow band LEDs. The results show that it is possible to obtain bilirubin and haemoglobin distribution maps and observe changes of chromophore parameter values over time by using a simple RGB imaging device. Water in vitro was detected by using differences in absorption at 450 nm and 950 nm, and 650 nm and 950 nm.
Application of fluorescence spectroscopy and multispectral imaging for non-invasive estimation of GFP transfection efficiency
Electroporation and ultrasound induced sonoporation has been showed to induce plasmid DNA transfection to the mice tibialis cranialis muscle. It offers new prospects for gene therapy and cancer treatment. However, numerous experimental data are still needed to deliver the plausible explanation of the mechanisms governing DNA electro- or sono-transfection, as well as to provide the updates on transfection protocols for transfection efficiency increase. In this study we aimed to apply non-invasive optical diagnostic methods for the real time evaluation of GFP transfection levels at the reduced costs for experimental apparatus and animal consumption. Our experimental set-up allowed monitoring …
Fluorescence spectroscopy for estimation of anticancer drug sonodestruction in vitro
The effect of ultrasound exposure on bleomycin fluorescence and pharmacological properties is studied. Bleomycin was treated by ultrasound for 7 min. Bleomycin fluorescence was measured during ultrasound exposure by means of fiber-optic spectrometry. Cell colony test was used to evaluate blemycin cytotoxity before and after ultrasound exposure.
Evaluation of Skin Melanoma in Spectral Range 450-950 nm Using Principal Component Analysis
Diagnostic potential of principal component analysis (PCA) of multi-spectral imaging data in the wavelength range 450-950 nm for distant skin melanoma recognition is discussed. Processing of the measured clinical data by means of PCA resulted in clear separation between malignant melanomas and pigmented nevi.
pEGFP transfection into murine skeletal muscle by electrosonoporation
In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can affect the plasmid DNA transfection to mice tibialis cranialis muscle. Multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) fluorescence, providing information on location and duration of EGFP expression. We found that electrosonoporation, commonly enhancing pDNA transfection in vitro , had no positive effect on EGFP transfection efficiency increase in vivo with respect to electroporation alone. We presume that this may be associated w…
Towards single snapshot multispectral skin assessment
Skin assessment technology based on comparative analysis of single-pixel RGB signal values at poly-chromatic illumination has been proposed. Multi-spectral imaging information from a single snapshot RGB image data set with the inter-channel crosstalk correction can be extracted this way. Proof-of-concept evaluations and measurement results are presented and discussed. Potential of bi-chromatic illumination for skin hemoglobin mapping during arterial occlusion test has been demonstrated.
Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies.
We highlight the options available for noninvasive optical diagnostics of reporter gene expression in mouse tibialis cranialis muscle. An in vivo multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) expression, providing information on location and duration of EGFP expression and allowing quantification of EGFP expression levels. For EGFP coding plasmid (pEGFP-Nuc Vector, 10 μg/50 ml 10 μg/50 ml ) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monit…
Multi-spectral skin imaging by a consumer photo-camera
The possibilities to perform multi-band spectral imaging by means of a consumer color camera without external filters have been studied. Images at up to 6 spectral bands may be extracted from a single color image after appropriate signal processing. The proposed technique was tested in pilot measurements of in-vivo skin hemoglobin maps and laser-excited autofluorescence images.
Multispectral imaging of pigmented and vascular cutaneous malformations: the influence of laser treatment
The paper investigates influence and efficacy of laser therapy on pigmented and vascular cutaneous malformations by multispectral imaging technique. Parameter mapping of skin pigmented and vascular lesions and monitoring of the laser therapy efficacy are performed by multispectral imaging in wavelength range 450-700nm by scanning step - 10nm. Parameter maps of the oxyhemoglobin deoxyhemoglobin and melanin derived from the images are presented. Possibility of laser therapy efficacy monitoring by comparison of the parameter maps before and after laser treatment has been demonstrated. As both cutaneous pigmented and vascular malformations are commonly found lesions, the parameter mapping would…
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