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Accurate and timely assessment of burn wound severity is a critical component of wound management and has implications related to course of treatment. While most superficial burns and full thickness burns are easily diagnosed through visual inspection, burns that fall between these extremes are challenging to classify based on clinical appearance. Because of this, appropriate burn management may be delayed, increasing the risk of scarring and infection. Here we present an investigation that employs spatial frequency domain imaging (SFDI) and laser speckle imaging (LSI) as non-invasive technologies to characterize in-vivoburn severity. We used SFDI and LSI to investigate controlled burn wounds of graded severity in a Yorkshire pig model. Burn wounds were imaged starting at one hour after the initial injury and daily at approximately 24, 48 and 72 hours post burn. Biopsies were taken on each day in order to correlate the imaging data to the extent of burn damage as indicated via histological analysis. Changes in reduced scattering coefficient and blood flow could be used to categorize burn severity as soon as one hour after the burn injury. The results of this study suggest that SFDI and LSI information have the potential to provide useful metrics for quantifying the extent and severity of burn injuries.

One of the major constraints facing laser speckle imaging for blood-flow measurement is reliable measurement of the correlation time (τ(C)) of the back-scattered light and, hence, the blood's speed in blood vessels. In this Letter, we present a new model expression for integrated speckle contrast, which accounts not only for temporal integration but spatial integration, too, due to the finite size of the pixel of the CCD camera; as a result, we find that a correction factor should be introduced to the measured speckle contrast to properly determine τ(C); otherwise, the measured blood's speed is overestimated. Experimental results support our theoretical model.

Laser Speckle Contrast Imaging (LSCI) is an optical technique used to generate blood flow maps with high spatial and temporal resolution. It is well known that in LSCI, the speckle size must exceed the Nyquist criterion to maximize the speckle's pattern contrast. In this work, we study experimentally the effect of speckle-pixel size ratio not only in dynamic speckle contrast, but also on the calculation of the relative flow speed for temporal and spatial analysis. Our data suggest that the temporal LSCI algorithm is more accurate at assessing the relative changes in flow speed than the spatial algorithm.

Background and Objective: Spatial Frequency Domain Imaging (SFDI) is a non-contact wide-field optical imaging technology currently being developed to investigate the feasibility of quantitative non-invasive evaluation of burn wound severity in a rat model. Our objective is to determine the potential of SFDI for mapping quantitative changes in spatially resolved tissue oxygen saturation and water concentration may be indicative of burn wound severity, healing, and further complications. In this portion of the investigation, we focus on the development of a rat burn model and the acute response of tissue to burn wounds. Study Design/Materials and Methods: A controlled burn protocol involving a heated brass comb was applied to 6 rats. Imaging was acquired at 17 evenly spaced wavelengths in the near-infrared from 650 to 970 nm. Over the course of the 3 hour post-burn period, we were able to map quantitative changes in spatially resolved chromophores. Burn severities were verified post-experiment using standard H and E histology and optical microscopy. Results/Conclusion: In total, we were able to induce 12 superficial-partial thickness burns, 8 deep-partial thickness burns, and 4 full thickness burns in our rat models. While several tissue chromophores were tracked, we found that changes in oxygen saturation and water concentration to be sensitive indicators of burn severity. Future work will include additional longitudinal studies over a period of days in order to investigate which parameters are correlated to tissue healing. © 2013 SPIE.

Port-wine stain (PWS) birthmarks are one class of benign congenital vascular malformation. Laser therapy is the most successful treatment modality of PWS. Unfortunately, this approach has limited efficacy, with only 10% of patients experiencing complete blanching of the PWS. To address this problem, several research groups have developed technologies and methods designed to study treatment outcome and improve treatment efficacy. This article reviews seven optical imaging techniques currently in use or under development to assess treatment efficacy, focusing on: reflectance spectrophotometers/tristimulus colorimeters; laser Doppler flowmetry and laser Doppler imaging; cross-polarized diffuse reflectance colour imaging system; reflectance confocal microscopy; optical coherence tomography; spatial frequency domain imaging; and laser speckle imaging.

The functionality of vascular networks within implanted prevascularized tissues is difficult to assess using traditional analysis techniques, such as histology. This is largely due to the inability to visualize hemodynamics in vivo longitudinally. Therefore, we have developed dynamic imaging methods to measure blood flow and hemoglobin oxygen saturation in implanted prevascularized tissues noninvasively and longitudinally. Using laser speckle imaging, multispectral imaging, and intravital microscopy, we demonstrate that fibrin-based tissue implants anastomose with the host (severe combined immunodeficient mice) in as short as 20h. Anastomosis results in initial perfusion with highly oxygenated blood, and an increase in average hemoglobin oxygenation of 53%. However, shear rates in the preformed vessels were low (20.8±12.8s-1), and flow did not persist in the vast majority of preformed vessels due to thrombus formation. These findings suggest that designing an appropriate vascular network structure in prevascularized tissues to maintain shear rates above the threshold for thrombosis may be necessary to maintain flow following implantation. We conclude that wide-field and microscopic functional imaging can dynamically assess blood flow and oxygenation in vivo in prevascularized tissues, and can be used to rapidly evaluate and improve prevascularization strategies. © Copyright 2012, Mary Ann Liebert, Inc.

Early neurovascular coupling (NVC) changes in Alzheimer's disease can potentially provide imaging biomarkers to assist with diagnosis and treatment. Previous efforts to quantify NVC with intrinsic signal imaging have required assumptions of baseline optical pathlength to calculate changes in oxy- and deoxy-hemoglobin concentrations during evoked stimuli. In this work, we present an economical spatial frequency domain imaging (SFDI) platform utilizing a commercially available LED projector, camera, and off-the-shelf optical components suitable for imaging dynamic optical properties. The fast acquisition platform described in this work is validated on silicone phantoms and demonstrated in neuroimaging of a mouse model. © 2012 SPIE.

Spatial Frequency Domain Imaging (SFDI) is a non-contact imaging method that uses multiple frequency spatial illumination to generate two dimensional maps of tissue optical properties (absorption and reduced scattering) and chromophore concentrations. We present phantom validation and pilot clinical data of a deployed light-emitting diode (LED) based system. The system employs four LED wavelengths (658 nm, 730 nm, 850 nm, 970 nm) to quantitatively assess tissue health by measurement of common tissue constituents. Phantom validation results and maps of oxyhemoglobin, deoxy-hemoglobin, water content, reduced scattering, and surface topography will be presented for pilot studies assessing burn severity and efficacy of port wine stain treatment. © 2012 SPIE.

The traditional phase-resolved Doppler method demonstrates great success for in-vivo imaging of blood flow and blood vessels. However, the phase-resolved method always requires high phase stability of the system. In phase instable situations, the performance of the phase-resolved methods will be degraded. We propose a modified Doppler variance algorithm that is based on the intensity or amplitude value. Performances of the proposed algorithm are compared with traditional phase-resolved Doppler variance and color Doppler methods for both phase stable and phase instable systems. For the phase instable situation, the proposed algorithm demonstrates images without phase instability induced artifacts. In-vivo imaging of window-chamber hamster skin is demonstrated for phase instable situation with a spectrometer-based Fourier domain OCT system. A microelectromechanical systems (MEMS) based swept source OCT (SSOCT) system is also used to demonstrate the performance of the proposed method in a phase instable situation. The phase stability of the SSOCT system is analyzed. In-vivo imaging of the blood vessel of human skin is demonstrated with the proposed method and the SSOCT system. For the phase stable situation, the proposed algorithm also demonstrates comparable performance with traditional phase-resolved methods. In-vivo imaging of the human choroidal blood vessel network is demonstrated with the proposed method under the phase stable situation. Depth-resolved fine choroidal blood vessel networks are shown. © 2011 Optical Society of America.

Speckle-based methods have been developed to characterize tissue blood flow and perfusion. One such method, called modified laser speckle imaging (mLSI), enables computation of blood flow maps with relatively high spatial resolution. Although it is known that the sensitivity and noise in LSI measurements depend on image exposure time, a fundamental disadvantage of mLSI is that it does not take into account this parameter. In this work, we integrate the exposure time into the mLSI method and provide experimental support of our approach with measurements from an in vitro flow phantom.

Recent studies have demonstrated that topical application of glycerol on intact skin does not affect its optical scattering properties. Investigators from our research group recently revisited the use of dimethyl sulfoxide (DMSO) as an agent with optical clearing potential. We address the use of optical clearing to enhance quantitation of subsurface fluorescence emission. We employed both in vitro and in vivo model systems to study the effect of topical DMSO application on fluorescence emission. Our in vitro experiments performed on a tissue-simulating phantom suggest that DMSO-mediated optical clearing enables enhanced characterization of subsurface fluorophores. With topical DMSO application, a marked increase in fluorescence emission was observed. After 30 min, the fluorescence signal at the DMSO-treated site was 9× greater than the contralateral saline-treated site. This ratio increased to 13× at 105 min after agent application. In summary, DMSO is an effective optical clearing agent for improved fluorescence emission quantitation and warrants further study in preclinical in vivo studies. Based on outcomes from previous clinical studies on the toxicity profile of DMSO, we postulate that clinical application of DMSO as an optical clearing agent, can be performed safely, although further study is warranted. © 2010 World Scientific Publishing Company.

A common strategy to study breast cancer is the use of the preclinical model. These models provide a physiologically relevant and controlled environment in which to study both response to novel treatments and the biology of the cancer. Preclinical models, including the spontaneous tumor model and mammary window chamber model, are very amenable to optical imaging and to this end, we have developed a wide-field functional imaging (WiFI) instrument that is perfectly suited to studying tumor metabolism in preclinical models. WiFI combines two optical imaging modalities, spatial frequency domain imaging (SFDI) and laser speckle imaging (LSI). Our current WiFI imaging protocol consists of multispectral imaging in the near infrared (650-980 nm) spectrum, over a wide (7 cm x 5 cm) field of view. Using SFDI, the spatially-resolved reflectance of sinusoidal patterns projected onto the tissue is assessed, and optical properties of the tissue are determined, which are then used to extract tissue chromophore concentrations in the form of oxy-, deoxy-, and total hemoglobin concentrations, and percentage of lipid and water. In the current study, we employ Monte Carlo simulations of SFDI light propagation in order to characterize the penetration depth of light in both the spontaneous tumor model and mammary window chamber model. Preliminary results suggest that different spatial frequency and wavelength combinations have different penetration depths, suggesting the potential depth sectioning capability of the SFDI component of WiFI. © 2010 Copyright SPIE - The International Society for Optical Engineering.

We present a combined optical Doppler tomography/spectral Doppler imaging modality to quantitatively evaluate the dynamic blood circulation and the artery blockage before and after a localized ischemic stroke in a mouse model. Optical Doppler Tomography (ODT) combines the Doppler principle with optical coherence tomography for noninvasive localization and measurement of particle flow velocity in highly scattering media with micrometer scale spatial resolution. Spectral Doppler imaging (SDI) provides complementary temporal flow information to the spatially distributed flow information of Doppler imaging. Fast, repeated, ODT scans across an entire vessel were performed to record flow dynamic information with high temporal resolution of cardiac cycles. Spectral Doppler analysis of continuous Doppler images demonstrates how the velocity components and longitudinally projected flow-volume-rate change over time for scatters within the imaging volume using spectral Doppler waveforms. Furthermore, vascular conditions can be quantified with various Doppler-angle-independent flow indices. Non-invasive in-vivo mice experiments were performed to evaluate microvascular blood circulation of a localized ischemic stroke mouse model. © 2010 Copyright SPIE - The International Society for Optical Engineering.

Functional optical characterization of disease progression and response to therapy suffers from loss of spatial resolution and imaging depth due to scattering, impacting the ability of researchers to localize and quantify molecular processes.Here we report on the ability of dimethyl sulfoxide (DMSO) to reduce temporarily the optical scattering of skin. Data collected from in vitro phantom images and in vivo fluorescence images demonstrate the potential of this simple method to mitigate the blurring effects of scattering with topical application, which we expect will improve the accuracy and localization of in vivo molecular imaging studies. © 2009 SPIE.

During laser therapy of port wine stain (PWS) birthmarks, regions of persistent perfusion may exist. We hypothesize that such regions, which are not readily visible, exist even during laser surgery performed by highly experienced clinicians. The objective of this study was to use objective feedback to assess the acute vascular response to laser therapy. We have developed a clinic-friendly laser speckle imaging (LSI) instrument to provide the clinician with real-time images of blood flow during laser therapy. We acquired images from patients undergoing laser therapy of PWS birthmarks at Scripps Clinic and Beckman Laser Institute and Medical Clinic. We extracted blood flow maps from the acquired imaging data. Collectively, we have observed two regimes of patient response to therapy: 1) an immediate increase in perfusion within minutes after laser therapy; and 2) an overall decrease in blood perfusion approximately one hour after laser therapy, with distinct regions of persistent perfusion apparent in the majority of post-treatment blood-flow images. A comparison of blood flow in PWS and adjacent normal skin demonstrated that PWS blood flow can be greater than or sometimes equivalent to that of normal skin. Regions of persistent perfusion frequently exist immediately after laser therapy of PWS birthmarks. Existence of these regions may be correlated to the need for multiple treatment sessions to improve substantially PWS skin appearance. With the use of intraoperative LSI, immediate retreatment of these regions may improve the outcome of each session. © 2009 SPIE.

Background: Little is known about cross-language measurement equivalence of the job content questionnaire (JCQ) Purpose: The purposes of this study were to assess the extent of cross-language differential item functioning (DIF) of the 27 JCQ items in six languages (French, Dutch, Belgian-French, Belgian-Dutch (Flemish), Italian, and Swedish) from six European research centers and to test whether its effects on the scale-level mean comparisons among the centers were substantial or not. Method: A partial gamma coefficient method was used for statistical DIF analyses where the Flemish JCQ was the reference for other language versions. Additionally, equivalence between the Flemish and Dutch translations was subjected to a judgmental review. Results: On average, 36% to 39% of the total tested items appeared to be cross-language DIF items in the statistical analyses. The judgmental review indicated that half of the DIF items may be associated with translation difference. The impacts of the DIF items on the mean comparisons of the JCQ scales between the centers were non-trivial: underestimated skill discretion (Milan), underestimated decision authority (Leiden), underestimated psychological demands (Milan women), and incomparable coworker support (Gothenburg 95). Conclusion: Cross-language DIF of the JCQ among European countries should be considered in international comparative studies on psychosocial job hazards using JCQ scales. © 2009 International Society of Behavioral Medicine.

Doppler Optical Coherence Tomography (DOCT) imaging of in-vivo retinal blood flow was widely studied as efforts of research community to push this technology into clinic. Spectral Doppler imaging of DOCT has been demonstrated as a quantification method of in-vivo pulsatile retinal blood flow in human eye. This technology has the all the advantages inherited from OCT comparing to Doppler ultrasound. Comparing to normal spatial-distributed color Doppler imaging of DOCT, spectral Doppler imaging can reveal more haemodynamics details on the time dimension. Although resistance index (RI) of a micro-vascular can be measured in vivo from human retina, the clinical significance of RI easurements still needs to be investigated. In vitro experiment conduced with ultrasound has demonstrated the higher vascular resistance value is associated with the higher RI measured assuming the constant compliance of vascular tube. In this study, the rodent window-chamber model (RWCM) was used as a platform to investigate the RI change as the micro-vasculature response to laser irradiation. The higher RI was measured after the occlusion of two veins (should it be arterials) that was verified with laser speckle imaging in our preliminary experiment results. © 2009 SPIE.

An overview is presented of recent trends in coherent anti-Stokes Raman scattering (CARS) microscopy. We briefly discuss the influence of tissue scattering on the CARS signal, methods for controlling the CARS emission and prospects for surface-enhancement of the CARS radiation.

CARS microscopy is used to quantitatively investigate the process of skin optical clearing. Using glycerol and dimethyl-sulfoxide as the clearing agents, we find that tissue scattering is a highly nonlinear function of agent concentration. © 2007 Optical Society of America.

Port wine stain (PWS) birthmarks are progressive vascular malformations that occur in ∼12,000 live births per year in the United States. The majority (∼90%) of PWS birthmarks occur on the head and neck regions, and thus are difficult to conceal. The psychosocial development of individuals with PWS birthmarks is adversely affected. Facial PWS lesions have been associated with increased incidence of glaucoma and seizures. The progressive nature of PWS skin may be due to lack of neuronal regulation of blood vessel size. Progressive development of the PWS results in a darker appearance, soft tissue hypertrophy, nodularity, and overall further disfigurement. Current treatment options have significant limitations in terms of efficacy and risk. With laser therapy, a reduction in size and degree of redness of PWS skin occurs in ∼60% of treated patients. After ten treatment sessions, complete disappearance of the PWS occurs in only ∼10% of treated patients. To reduce the financial burden and potential risks of repeated treatments under general anesthesia, there is a need for innovative, personalized methods to maximize the reduction in PWS redness per treatment session. Without addressing this need, the overall efficacy of PWS laser therapy will remain variable, because treatment protocols will remain based primarily on the subjective impression and overall experience of the treating surgeon. To address this need, we propose use of laser speckle imaging (LSI) to provide real-time, quantitative feedback during laser surgery.