Lise Lyngsnes Randeberg

Characterization of vascular structures and skin bruises using hyperspectral imaging, image analysis and diffusion theory

Hyperspectral imaging, image analysis and diffusion theory were used to visualize skin vasculature and to monitor the development of fresh skin bruises. Bruises were inflicted in a porcine model, and the development of the hemorrhage was monitored using white light hyperspectral imaging (400-1000 nm). Hyperspectral images from human volunteers were also included in the study. Statistical image analysis was used to classify bruised regions and to visualize the skin vasculature. Biopsies were collected from the animals to reveal the true depth of the bruising. A three-layer diffusion model and an analytic hemoglobin transport model were used to model the reflectance spectra from the images. The results show that hyperspectral images contain depth information, and that the approximate depth and extent of bruises can be retrieved using a combination of statistical image analysis and diffusion theory. This technique also shows potential to visualize vascular structures in human skin.

Hyperspectral imaging of bruised skin

Bruises can be important evidence in legal medicine, for example in cases of child abuse. Optical techniques can be used to discriminate and quantify the chromophores present in bruised skin, and thereby aid dating of an injury. However, spectroscopic techniques provide only average chromophore concentrations for the sampled volume, and contain little information about the spatial chromophore distribution in the bruise. Hyperspectral imaging combines the power of imaging and spectroscopy, and can provide both spectroscopic and spatial information. In this study a hyperspectral imaging system developed by Norsk Elektro Optikk AS was used to measure the temporal development of bruised skin in a human volunteer. The bruises were inflicted by paintball bullets. The wavelength ranges used were 400 - 1000 nm (VNIR) and 900 - 1700 nm (SWIR), and the spectral sampling intervals were 3.7 and 5 nm, respectively. Preliminary results show good spatial discrimination of the bruised areas compared to normal skin. Development of a white spot can be seen in the central zone of the bruises. This central white zone was found to resemble the shape of the object hitting the skin, and is believed to develop in areas where the impact caused vessel damage. These results show that hyperspectral imaging is a promising technique to evaluate the temporal and spatial development of bruises on human skin.

Estimation of skin optical parameters for real-time hyperspectral imaging applications

Abstract. Hyperspectral imaging combines high spectral and spatial resolution in one modality. This imaging technique is a promising tool for objective medical diagnostics. However, to be attractive in a clinical setting, the technique needs to be fast and accurate. Hyperspectral imaging can be used to analyze tissue properties using spectroscopic methods, and is thus useful as a general purpose diagnostic tool. We combine an analytic diffusion model for photon transport with real-time analysis of the hyperspectral images. This is achieved by parallelizing the inverse photon transport model on a graphics processing unit to yield optical parameters from diffuse reflectance spectra. The validity of this approach was verified by Monte Carlo simulations. Hyperspectral images of human skin in the wavelength range 400–1000 nm, with a spectral resolution of 3.6 nm and 1600 pixels across the field of view (Hyspex VNIR-1600), were used to develop the presented approach. The implemented algorithm was found to output optical properties at a speed of 3.5 ms per line of image data. The presented method is thus capable of meeting the defined real-time requirement, which was 30 ms per line of data.The algorithm is a proof of principle, which will be further developed.

Monitoring of hexyl 5-aminolevulinate-induced photodynamic therapy in rat bladder cancer by optical spectroscopy

Monitoring of the tissue response to photodynamic therapy (PDT) can provide important information to help optimize treatment variables such as drug and light dose, and possibly predict treatment outcome. A urinary bladder cancer cell line (AY-27) was used to induce orthotopic transitional cell carcinomas (TCC) in female Fischer rats, and hexyl 5-aminolevulinate (HAL, 8 mM, 1 h)-induced PDT was performed on day 14 after instillation of the cancer cells (20 J/cm(2) fluence at 635 nm). In vivo optical reflectance and fluorescence spectra were recorded from bladders before and after laser treatment with a fiberoptic probe. Calculated fluorescence bleaching and oxygen saturation in the bladder wall were examined and correlated to histology results. Reflectance spectra were analyzed using a three-layer optical photon transport model. Animals with TCC treated with PDT showed a clear treatment response; decreased tissue oxygenation and protoporphyrin IX (PpIX) fluorescence photobleaching were observed. Histology demonstrated that 3 of 6 animals with treatment had no sign of the tumor 7 days after PDT treatment. The other 3 animals had significantly reduced the tumor size. The most treatment-responsive animals had the highest PpIX fluorescence prior to light irradiation. Thus, optical spectroscopy can provide useful information for PDT. The model has proved to be very suitable for bladder cancer studies.

Hyperspectral imaging of atherosclerotic plaques in vitro.

Vulnerable plaques constitute a risk for serious heart problems, and are difficult to identify using existing methods. Hyperspectral imaging combines spectral- and spatial information, providing new possibilities for precise optical characterization of atherosclerotic lesions. Hyperspectral data were collected from excised aorta samples (n = 11) using both white-light and ultraviolet illumination. Single lesions (n = 42) were chosen for further investigation, and classified according to histological findings. The corresponding hyperspectral images were characterized using statistical image analysis tools (minimum noise fraction, K-means clustering, principal component analysis) and evaluation of reflectance/fluorescence spectra. Image analysis combined with histology revealed the complexity and heterogeneity of aortic plaques. Plaque features such as lipids and calcifications could be identified from the hyperspectral images. Most of the advanced lesions had a central region surrounded by an outer rim or shoulder-region of the plaque, which is considered a weak spot in vulnerable lesions. These features could be identified in both the white-light and fluorescence data. Hyperspectral imaging was shown to be a promising tool for detection and characterization of advanced atherosclerotic plaques in vitro. Hyperspectral imaging provides more diagnostic information about the heterogeneity of the lesions than conventional single point spectroscopic measurements.

Hyperspectral imaging as a diagnostic tool for chronic skin ulcers

The healing process of chronic wounds is complex, and the complete pathogenesis is not known. Diagnosis is currently based on visual inspection, biopsies and collection of samples from the wound surface. This is often time consuming, expensive and to some extent subjective procedures. Hyperspectral imaging has been shown to be a promising modality for optical diagnostics. The main objective of this study was to identify a suitable technique for reproducible classification of hyperspectral data from a wound and the surrounding tissue. Two statistical classification methods have been tested and compared to the performance of a dermatologist. Hyperspectral images (400-1000 nm) were collected from patients with venous leg ulcers using a pushbroom-scanning camera (VNIR 1600, Norsk Elektro Optikk AS).Wounds were examined regularly over 4 - 6 weeks. The patients were evaluated by a dermatologist at every appointment. One patient has been selected for presentation in this paper (female, age 53 years). The oxygen saturation of the wound area was determined by wavelength ratio metrics. Spectral angle mapping (SAM) and k-means clustering were used for classification. Automatic extraction of endmember spectra was employed to minimize human interaction. A comparison of the methods shows that k-means clustering is the most stable method over time, and shows the best overlap with the dermatologist’s assessment of the wound border. The results are assumed to be affected by the data preprocessing and chosen endmember extraction algorithm. Results indicate that it is possible to develop an automated method for reliable classification of wounds based on hyperspectral data.

Applicability of diffusion approximation in analysis of diffuse reflectance spectra from healthy human skin

Measurement of diffuse reflectance spectra (DRS) is a common experimental approach for non-invasive determination of tissue optical properties, as well as objective monitoring of various tissue malformations. Propagation of light in scattering media is often treated in diffusion approximation (DA). The major advantage of this approach is that it leads to enclosed analytical solutions for tissues with layered structure, which includes human skin. Despite the fact that DA solutions were shown to be inaccurate near tissue boundaries, the practicality of this approach makes it quite popular, especially when attempting extraction of specific chromophore concentrations from measured DRS. In this study we analyze the discrepancies between DRS spectra as obtained by using the DA solutions for three-layer skin model and more accurate predictions from Monte Carlo (MC) modeling. Next, we analyze the artifacts which result from the above discrepancies when extracting the parameters of skin structure and composition by fitting the DA solutions to the MC spectra. The reliability and usefulness of such a fit is then tested also on measurements of seasonal changes in otherwise healthy human skin.

Hyperspectral imaging of bruises in the SWIR spectral region

Optical diagnostics of bruised skin might provide important information for characterization and age determination of such injuries. Hyperspectral imaging is one of the optical techniques that have been employed for bruise characterization. This technique combines high spatial and spectral resolution and makes it possible to study both chromophore signatures and -distributions in an injury. Imaging and spectroscopy in the visible spectral range have resulted in increased knowledge about skin bruises. So far the SWIR region has not been explored for this application. The main objective of the current study was to characterize bruises in the SWIR wavelength range. Hyperspectral images in the SWIR (950-2500nm ) and VNIR (400-850nm) spectral range were collected from 3 adult volunteers with bruises of known age. Data were collected over a period of 8 days. The data were analyzed using spectroscopic techniques and statistical image analysis. Preliminary results from the pilot study indicate that SWIR hyperspectral imaging might be an important supplement to imaging in the visible part of the spectrum. The technique emphasizes local edema and gives a possibility to visualize features that cannot easily be seen in the visible part of the spectrum.

Real-Time Noise Removal for Line-Scanning Hyperspectral Devices Using a Minimum Noise Fraction-Based Approach

Processing line-by-line and in real-time can be convenient for some applications of line-scanning hyperspectral imaging technology. Some types of processing, like inverse modeling and spectral analysis, can be sensitive to noise. The MNF (minimum noise fraction) transform provides suitable denoising performance, but requires full image availability for the estimation of image and noise statistics. In this work, a modified algorithm is proposed. Incrementally-updated statistics enables the algorithm to denoise the image line-by-line. The denoising performance has been compared to conventional MNF and found to be equal. With a satisfying denoising performance and real-time implementation, the developed algorithm can denoise line-scanned hyperspectral images in real-time. The elimination of waiting time before denoised data are available is an important step towards real-time visualization of processed hyperspectral data. The source code can be found at http://www.github.com/ntnu-bioopt/mnf. This includes an implementation of conventional MNF denoising.

In vivo hyperspectral imaging of traumatic skin injuries in a porcine model

Studies of immediate skin reactions are important to understand the underlying biological mechanisms involved in traumatic or chemical damage to the skin. In this study the spatial and spectral information provided by hyperspectral images was used to identify and characterize non-penetrating skin injuries in a porcine model. A hyperspectral imaging system (Hyspex, Norsk Elektro Optikk AS) was used to monitor the temporal development of minor skin injuries in an anesthetized Norwegian domestic pig. Hyperspectral data were collected in the wavelength range 400-1000 nm (VNIR), with a spectral sampling interval of 3.7 nm. The measurements were initiated immediately after inflicting the injury, and were repeated at least five times at each site with irregular frequency. The last measurement was performed 4 hours after injury. Punch biopsies (5 mm), were collected from adjacent normal skin, and at the center and the margin of each injury. The study was approved by the national animal research authority. The hyperspectral data were analyzed with respect to oxy- and deoxyhemoglobin, and erythema index. The skin biopsies were examined to determine the extent of skin damage in the bruised zones. Preliminary results show that hyperspectral imaging allows discrimination between traumatized skin and normal skin in an early phase. The extent and location of the hemorrhages can be determined from hyperspectral images. These findings might contribute to a better understanding of immediate skin reactions to minor trauma, and thereby the development of a better diagnostic modality for non-penetrating skin injuries in forensic medicine.