Search

A critical need exists for early, accurate diagnosis of burn wound severity to help identify the course of treatment and outcome of the wound. Laser speckle imaging (LSI) is a promising blood perfusion imaging approach, but it does not account for changes in tissue optical properties that can occur with burn wounds, which are highly dynamic environments. Here, we studied optical property dynamics following burn injury and debridement and the associated impact on interpretation of LSI measurements of skin perfusion. We used spatial frequency domain imaging (SFDI) measurements of tissue optical properties to study the impact of burn-induced changes in these properties on LSI measurements. An established preclinical porcine model of burn injury was used (n = 8). SFDI and LSI data were collected from burn wounds of varying severity. SFDI measurements demonstrate that optical properties change in response to burn injury in a porcine model. We then apply theoretical modeling to demonstrate that the measured range of optical property changes can affect the interpretation of LSI measurements of blood flow, but this effect is minimal for most of the measured data. Collectively, our results indicate that, even with a dynamic burn wound environment, blood-flow measurements with LSI can serve as an appropriate strategy for accurate assessment of burn severity.

PDF file - 181KB, Compound nevus. (Upper row) MPM images of normal skin (adjacent to nevus) showing (a) keratinocytes in the stratum spinosum at a depth of 30 microm, (b) basal cells (green) surrounding dermal papilla (blue) - 50 microm deep, (c) collagen fibers in the dermis - 75 microm deep. (Lower row) MPM images of the nevus showing pigmented keratinocytes in the stratum spinosum at a depth of 30 microm (d), basal cells (green) surrounding dermal papilla (blue) at depths of 50 (e) and 75 microm (f). Scale bar is 40 microm. Inset shows the clinical image of the nevus (scale bar is 1 mm).

PDF file - 178KB, Melanoma. (a) Clinical image. (b) Histology image of the lesion showing pagetoid spread in the upper epidermal layers (arrows). (c-e) MPM images of epidermal layers showing advanced melanocytic dendrites (red arrows) and pleomorphic cells/pagetoid spread (yellow arrows) in upper epidermal layers at depths of 40 microm (c), 45 microm, (d) and 50 microm (e). Scale bar is 20 microm.

PDF file - 171KB, Dysplastic nevus. (a) Clinical image (b) Histology image of the lesion showing melanocytes advanced in the upper epidermal layers (arrows). (c-d) MPM images of keratinocytes in epidermal layers at 25 microm (c) and 35 microm (d). Images show presence of melanocytic dendrites in the spinosum layer of epidermis (arrows). (e) MPM image of the basal layer showing cells with enlarged nuclei (arrows). Scale bar is 20 microm.

Clinical studies have demonstrated that epidermal pigmentation level can affect cerebral oximetry measurements. To evaluate the robustness of these devices, we have developed a phantom-based test method that includes an epidermis-simulating layer with several melanin concentrations and a 3D-printed cerebrovascular module. Measurements were performed with neonatal, pediatric and adult sensors from two commercial oximeters, where neonatal probes had shorter source-detector separation distances. Referenced blood oxygenation levels ranged from 30 to 90%. Cerebral oximeter outputs exhibited a consistent decrease in saturation level with simulated melanin content; this effect was greatest at low saturation levels, producing a change of up to 15%. Dependence on pigmentation was strongest in a neonatal sensor, possibly due to its high reflectivity. Overall, our findings indicate that a modular channel-array phantom approach can provide a practical tool for assessing the impact of skin pigmentation on cerebral oximeter performance and that modifications to algorithms and/or instrumentation may be needed to mitigate pigmentation bias. Funding: FDA Office of Minority Health and Health Equity and FDA Perinatal Health Center of Excellence

Spatial frequency domain imaging (SFDI) is a wide-field diffuse optical imaging technique for separately quantifying tissue reduced scattering (μs ' ) and absorption (μa) coefficients at multiple wavelengths, providing wide potential utility for clinical applications such as burn wound characterization and cancer detection. However, measured μs ' and μa can be confounded by absorption from melanin in patients with highly pigmented skin. This issue arises because epidermal melanin is highly absorbing for visible wavelengths and standard homogeneous light-tissue interaction models do not properly account for this complexity. Tristimulus colorimetry (which quantifies pigmentation using the L * "lightness" parameter) can provide a point of comparison between μa, μs ' , and skin pigmentation.We systematically compare SFDI and colorimetry parameters to quantify confounding effects of pigmentation on measured skin μs ' and μa. We assess the correlation between SFDI and colorimetry parameters as a function of wavelength.μs ' and μa from the palm and ventral forearm were measured for 15 healthy subjects with a wide range of skin pigmentation levels (Fitzpatrick types I to VI) using a Reflect RS® (Modulim, Inc., Irvine, California) SFDI instrument (eight wavelengths, 471 to 851 nm). L * was measured using a Chroma Meter CR-400 (Konica Minolta Sensing, Inc., Tokyo). Linear correlation coefficients were calculated between L * and μs ' and between L * and μa at all wavelengths.For the ventral forearm, strong linear correlations between measured L * and μs ' values were observed at shorter wavelengths (R 0.92 at ≤659 nm), where absorption from melanin confounded the measured μs ' . These correlations were weaker for the palm (R 0.59 at ≤659 nm), which has less melanin than the forearm. Similar relationships were observed between L * and μa.We quantified the effects of epidermal melanin on skin μs ' and μa measured with SFDI. This information may help characterize and correct pigmentation-related inaccuracies in SFDI skin measurements.

Background Burns are devastating injuries, carry significant morbidity, and require long-term treatment or multiple reconstructive procedures. Wound healing and secondary insults caused by burn wound conversion are amendable to therapeutic intervention, where ischemia has been cited as one of the major factors (Dermatol Surg. 2008;34:1159-1169). Halting injury progression in the zone of stasis is crucial as conversion creates increased burn surface area and depth, leading to local and systemic consequences (J Burns Wounds. 2006;5:e2). Oxygen-carrying micro/nanobubbles, MNB(O2), offer a novel technology that can be used to effectively deliver oxygen to burn wounds and potentially counteract burn wound ischemia. Methods Topical irrigation with MNB(O2) of full-thickness burn wounds on a rodent model (n = 3) was compared against saline-treated controls (n = 3). Tissue structure (reduced scattering coefficient, μs'), oxyhemoglobin concentration (cHbO2), and tissue perfusion were quantified over the course of 28 days through spatial frequency domain imaging and laser speckle imaging. Histological samples taken at the end of the experiment were examined for evidence of wound healing. Results Findings in this preliminary study showed hastened healing with significant differences in spatial frequency domain imaging-measured μs' during wound healing (days 11-28) in MNB(O2) group. The healing "tipping point" seemed to occur at days 9 to 11 with increased collagen organization and increased cHbO2 occurring around that period confirming the gross healing improvements observed. In addition, histological evidence indicated that only the MNB(O2) burns had reached the remodeling phase by the end of 28-day study period. Conclusions These preliminary findings propose the potential of MNB(O2) as a topical method for improving burn wound healing.

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.

While clinical examination is needed for burn severity diagnosis, several emerging technologies aim to quantify this process for added objectivity. Accurate assessments become easier after burn progression, but earlier assessments of partial thickness burn depth could lead to earlier excision and grafting and subsequent improved healing times, reduced rates of scarring/infection, and shorter hospital stays. Spatial Frequency Domain Imaging (SFDI), Laser Speckle Imaging (LSI) and thermal imaging are three non-invasive imaging modalities that have some diagnostic ability for noninvasive assessment of burn severity, but have not been compared in a controlled experiment. Here we tested the ability of these imaging techniques to assess the severity of histologically confirmed graded burns in a swine model. Controlled, graded burn wounds, 3 cm in diameter were created on the dorsum of Yorkshire pigs (n=3, 45–55kg) using a custom-made burn tool that ensures consistent pressure has been employed by various burn research groups. For each pig, a total of 16 burn wounds were created on the dorsal side. Biopsies were taken for histological analysis to verify the severity of the burn. Clinical analysis, SFDI, LSI and thermal imaging were performed at 24 and 72 hours after burn to assess the accuracy of each imaging technique. In terms of diagnostic accuracy, using histology as a reference, SFDI (85%) and clinical analysis (83%) performed significantly better that LSI (75%) and thermography (73%) 24 hours after the burn. There was no statistically significant improvement from 24 to 72 hours across the different imaging modalities. These data indicate that these imaging modalities, and specifically SFDI, can be added to the burn clinicians’ toolbox to aid in early assessment of burn severity.

We propose a new image sensor based on multi-aperture optics and multi-tap charge modulators for simultaneous multi-band spatial frequency domain imaging (SFDI) and blood flow mapping by multi-exposure laser speckle contrast imaging (ME-LSCI). Wavelength-multiplexed imaging implemented by attaching different bandpass filters to each lens enables to perform multi-band SFDI and ME-LSCI simultaneously. A prototype camera for 4-band SFDI (568, 668, 734, 846nm) and single-band LSCI (780nm) was built based on a commercial CMOS camera with enhanced NIR sensitivity to demonstrate the concept and tested using a rat model of graded burns.

While visual clinical impression is the standard of care for burn severity assessment, accuracy improves after the injury progresses, leading to patients waiting days for a diagnosis. Spatial Frequency Domain Imaging (SFDI) has shown the potential to assess burn grade as early as 24 hours post-burn. Here, we compared SFDI and Laser Speckle Imaging (LSI) at 24 and 72 hours after burn injury to clinical burn severity diagnosis. Three patients with burns ranging in severity were imaged. Both techniques showed an ability to categorize burn severity 24 hours after the initial injury, well before the outcome was determined by a clinician at 72 hours or later.

Spatial frequency domain imaging (SFDI) is a wide-field imaging technique that provides quantitative tissue optical property maps. We describe a compact, multi-aperture SFDI imager for burn severity evaluation. This was constructed using a CMOS sensor subdivided into multiple regions, each having a bandpass filter and objective lens. This compound-eye design enables simultaneous image acquisition at multiple wavelengths. The imager was evaluated using a rat burn model and compared to conventional SFDI devices that acquire wavelength data sequentially. The burns showed a decrease in the reduced scattering coefficient that is consistent with previously reported changes in full thickness burns.

Spatial Frequency Domain Imaging (SFDI) is a non-invasive diffuse optical imaging technology that quantifies tissue optical properties by projecting spatially modulated light onto a region of interest. By detecting and fitting the diffuse reflectance to a light transport model, tissue absorption (μa) and scattering (μs') coefficients are calculated. However, when measuring skin, bulk μa and μs' from a homogeneous light transport model may not correspond with the actual properties of individual skin layers, especially for highly pigmented skin. To obtain physiologically accurate skin optical property values, we propose an iterative method based on a two- layered Monte Carlo model. We initially assume that μs' in darker skin is the same as that in lighter skin, and set the epidermal a as the free parameter when fitting the diffuse reflectance. To test this algorithm, we analyzed data from the forearms of 6 subjects having various levels of pigmentation, at 8 visible-to-near-infrared wavelengths. We compared the measured reflectance to both the homogeneous model and our layered model to quantify fit accuracy. At 471 nm and 526 nm for patients of Fitzpatrick skin types IV-VI (relatively dark skin), the two-layered model provided a 10-20% improvement in fit to the AC reflectance as a function of spatial frequency, compared to the homogeneous model. These improved fits yielded epidermal absorption coefficients that were notably higher than the bulk μa from the homogeneous model. Fitting the extracted epidermal μa to a melanin extinction spectrum1 enabled estimation of the melanin concentration in the epidermis.

Adequate resuscitation is critical for burn wound patients where the affected total body surface area (TBSA) exceeds 20%, as fluid loss leads to organ failure, shock, and patient mortality. Protocols have been developed to combat fluid loss, however excess fluids can also lead to compartment syndromes. Here, we used spatial frequency domain imaging (SFDI), a non-contact wide field imaging technique to measure in-vivo water changes in a 40 % TBSA porcine burn model. In this pilot study one pig received intravenous fluids according to the Parkland formula on top of enteral fluid resuscitation, while a second pig received no fluids during the experiment. Unburned regions of skin were imaged with SFDI from 3 to 22 hours post-burn. This imaging technique uses structured illumination, projected at multiple spatial frequencies and wavelengths, to measure tissue reflectance, which is used to obtain the reduced scattering and absorption coefficients. Water fraction is based upon absorption in the near-infrared spectrum at 971 nm, where water has a high extinction coefficient. SFDI measurements of superficial tissue hydration showed increased water fraction for the pig that received fluid resuscitation (+17%), and a decrease for the pig that received no fluids (-5%). Analysis showed decreased scattering measured in the pig that received fluids (+13%), suggesting increased interstitial water pressure. These preliminary results are consistent with systemic hydration, as estimated based on CT-measured subcutaneous fat thickness. SFDI may, therefore offer the possibility of non-invasively monitoring fluid resuscitation of large TBSA patients.

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.

While visual assessment by a clinician is the standard of care for burn severity evaluations, new technologies at various stages of development are attempting to add objectivity to this practice by quantifying burn severity. Assessment accuracy generally improves after the burn injury has progressed, but early assessments that correctly identify superficial partial and deep partial burns have the potential to lead to more prompt treatments and shorter recovery times. To date, Spatial Frequency Domain Imaging (SFDI) has only been used in animal models of burns, but has shown the potential to categorize burns accurately at earlier time points. Here we examine the potential for SFDI to assess burn severity in clinical patients. We also utilize Laser Speckle Imaging (LSI), an FDA cleared non-invasive imaging technology that typically measures blood perfusion in order to evaluate burns in clinical patients. We present a case series of two patients, both with partial thickness burns of varying severity. Partial thickness burns are often difficult for clinicians to categorize based on visual appearance alone. SFDI and LSI were both performed on each patient at approximately 24 and 72 h after their respective burn incidents. Each technique was able to render spatially resolved information that enabled improved assessment accuracy for each burn. This represents the first publication of SFDI applied to clinical burn patients after being successfully utilized in animal models, and highlights the potential for SFDI as a feasible tool for the timely categorization of burn severity. Keywords: Burns, Clinical, Humans, Spatial Frequency Domain Imaging, Laser Speckle Imaging

We have optimized our prior phantom-based test method for cerebral oximetry performance using a new 3D-printed cerebrovascular module (CVM). In addition, we have and added a surrogate for a thin perfused scalp layer to evaluate perfusion- related confounding factors. The new CVM’s optical properties better represented biological tissue and also incorporated a water-mimicking dye. The modular phantom also included biologically relevant scalp/skull and cerebrospinal fluid (CSF) layers. Performance testing of two commercially available clinical oximeters with the modified CVM over a range of oxygen saturation levels illustrates the utility of our solid phantom-based approach for standardized cerebral oximeter performance assessment.

Delayed graft function occurs in 40% of kidney transplantation cases, and is not apparent until after surgery has concluded. To understand changes in donor kidney tissue before transplantation, spatial frequency domain imaging (SFDI) was used to measure absorption and reduced scattering coefficients in excised porcine kidneys as they underwent a standard period of cold ischemic time (CIT). SFDI indicated an increase in reduced scattering and absorption. The scattering b parameter increased until 24 hours, then returned towards baseline. These results suggest that continued investigation may lead to the development of a noninvasive assessment for graft viability.

Spatial frequency domain imaging (SFDI), a noncontact imaging technique using structured illumination with multiple wavelengths, has proved feasible for characterizing burns of different severities induced in porcine dorsum. However, translation to human clinical subjects may be subject to variation in tissue optical properties due to different skin types or body parts. To interrogate this potential difference in optical properties, SFDI was used to measure the reduced scattering coefficient (μs’ ) in patients of varying Fitzpatrick skin type groups at 10 anatomical locations on the body. These scattering values were compared against previously measured average μs’ values for burns in a porcine model. At 851nm, superficial partialthickness burns had a 12.6% higher μs’ than unburned porcine dorsum. Deep partial-thickness and had 12.9% higher average μs’ than unburned porcine dorsum, while full-thickness burns had 2.5% higher average μs’ than unburned porcine dorsum. The variation seen in scattering values at 851nm at one anatomical location (dorsal forearm) for the different skin type groups showed a 6.3% lower average μs’ in group 3 (skin types VVI) compared to group 2 (skin types III-IV). Average scattering values were 3.8% lower in group 1 (skin types I-II) compared to group 2 (skin types III-IV). The variation of μs’ at 851nm across multiple anatomical locations for the same skin type group (Group 1) was a 23% difference from the anatomical location with the highest average μs’ at 851nm (forehead) to the lowest (shin). These findings indicate that further consideration of skin type and location will be integral to the translation of classification techniques based μs’ measurements of porcine dorsum to human patients.

Herein, to investigate a new diagnostic method for Meibomian gland dysfunction (MGD) induced by eyelid inflammation, optical properties and deoxy-hemoglobin (Hb) concentrations in rodent eyelid tissues, including Meibomian glands(MGs), were measured using spatial frequency domain imaging (SFDI). Complete Freund’s adjuvant solutions were injected into the eyelid margins of Sprague–Dawley rats to induce MGD. After three weeks, the optical properties and Hb of the MG and non-MG regions of the eyelids were measured ex–vivo using an SFDI system. The comparison of Hb showed that the MGD group exhibited significantly higher values than those of the control group in both regions. The optical properties at 730 and 850 nm for the MG regions in the MGD group were significantly different from those in the control group. In addition, the 630 nm absorption coefficients of both regions were significantly higher in the MGD group than in the control group. Thus, the SFDI technique can detect the increased Hb concentration and changes in the optical properties of the eyelids due to inflammatory MGD in a noncontact manner and has the potential to be used as a novel quantitative diagnostic method for the occurrence of MGD.

Introduction While visual assessment is the standard for burn severity evaluation, new technologies are attempting to increase objectivity. Burn depth assessment accuracy improves over time as the appearance changes, but earlier diagnosis potentially leads to prompt treatment and shorter recovery. Laser Speckle Imaging (LSI), FDA approved for grading burns, is typically done 72hrs after injury. Spatial Frequency Domain Imaging (SFDI), a novel technology only tested in animal burn models, uses multiple wavelengths of patterned light to quantify tissue absorption (hemodynamics) and scattering (structure). SFDI has shown the potential to differentiate burns as early as 24hrs postburn by measuring scattering (µs’) changes due to collagen damage. Here, we examine SFDI burn severity assessment in clinical patients, and compare against LSI and clinical assessment. Methods 5 burn cases were imaged with LSI and SFDI approximately 24 and 72hrs postburn. The clinician was blinded to imaging results and made diagnoses on their standard of care. Results One patient received no surgical intervention, one received excision and xenograft, and two received excision and autograft. An autograft was recommended for one patient, but not performed due to preexisting conditions. SFDI measurements at 24hrs showed low µs’ values in regions that eventually required autografts, and high µs’ in burns that did not. While clinical decisions were made no earlier than 72hrs postburn, µs’ maps at 24hrs postburn were able to illustrate severity in these regions. In 2 of 3 autograft cases, LSI was unable to determine the severity until 72hrs postburn. Conclusions Here, we compared SFDI to LSI in a clinical setting to quantify burn wound severity. Similar to animal models, SFDI was demonstrated to accurately characterize severity earlier than the perfusion-based LSI. This is likely due to sensitivity of µs’ measured by SFDI to the structural changes in damaged collagen (i.e., zone of coagulation) that occur immediately postburn.

Author(s): Ponticorvo, Adrien; Burmeister, David M; Rowland, Rebecca; Baldado, Melissa; Kennedy, Gordon T; Saager, Rolf; Bernal, Nicole; Choi, Bernard; Durkin, Anthony J | Abstract: The current standard for diagnosis of burn severity and subsequent wound healing is through clinical examination, which is highly subjective. Several new technologies are shifting focus to burn care in an attempt to help quantify not only burn depth but also the progress of healing. While accurate early assessment of partial thickness burns is critical for dictating the course of treatment, the ability to quantitatively monitor wound status over time is critical for understanding treatment efficacy. SFDI and LSI are both non-invasive imaging modalities that have been shown to have great diagnostic value for burn severity, but have yet to be tested over the course of wound healing.In this study, a hairless rat model (n = 6, 300-450 g) was used with a four pronged comb to create four identical partial thickness burns (superficial n = 3 and deep n = 3) that were used to monitor wound healing over a 28 days period. Weekly biopsies were taken for histological analysis to verify wound progression. Both SFDI and LSI were performed weekly to track the evolution of hemodynamic (blood flow and oxygen saturation) and structural (reduced scattering coefficient) properties for the burns.LSI showed significant changes in blood flow from baseline to 220% in superficial and 165% in deep burns by day 7. In superficial burns, blood flow returned to baseline levels by day 28, but not for deep burns where blood flow remained elevated. Smaller increases in blood flow were also observed in the surrounding tissue over the same time period. Oxygen saturation values measured with SFDI showed a progressive increase from baseline values of 66-74% in superficial burns and 72% in deep burns by day 28. Additionally, SFDI showed significant decreases in the reduced scattering coefficient shortly after the burns were created. The scattering coefficient progressively decreased in the wound area, but returned towards baseline conditions at the end of the 28 days period. Scattering changes in the surrounding tissue remained constant despite the presence of hemodynamic changes.Here, we show that LSI and SFDI are capable of monitoring changes in hemodynamic and scattering properties in burn wounds over a 28 days period. These results highlight the potential insights that can be gained by using non-invasive imaging technologies to study wound healing. Further development of these technologies could be revolutionary for wound monitoring and studying the efficacy of different treatments. Lasers Surg. Med. 49:293-304, 2017. © 2017 Wiley Periodicals, Inc.

There is a need for noninvasive, quantitative methods to characterize wound healing in the context of longitudinal investigations related to regenerative medicine. Such tools have the potential to inform the assessment of wound status and healing progression and aid the development of new treatments. We employed spatial frequency domain imaging (SFDI) to characterize the changes in optical properties of tissue during wound healing progression in a porcine model of split-thickness skin grafts and also in a model of burn wound healing with no graft intervention. Changes in the reduced scattering coefficient measured using SFDI correlated with structural changes reported by histology of biopsies taken concurrently. SFDI was able to measure spatial inhomogeneity in the wounds and predicted heterogeneous healing. In addition, we were able to visualize differences in healing rate, depending on whether a wound was debrided and grafted, versus not debrided and left to heal without intervention apart from topical burn wound care. Changes in the concentration of oxy- and deoxyhemoglobin were also quantified, giving insight into hemodynamic changes during healing.

Accurate assessment of burn severity is critical for wound care and the course of treatment. Delays in classification translate to delays in burn management, increasing the risk of scarring and infection. To this end, numerous imaging techniques have been used to examine tissue properties to infer burn severity. Spatial frequency-domain imaging (SFDI) has also been used to characterize burns based on the relationships between histologic observations and changes in tissue properties. Recently, machine learning has been used to classify burns by combining optical features from multispectral or hyperspectral imaging. Rather than employ models of light propagation to deduce tissue optical properties, we investigated the feasibility of using SFDI reflectance data at multiple spatial frequencies, with a support vector machine (SVM) classifier, to predict severity in a porcine model of graded burns. Calibrated reflectance images were collected using SFDI at eight wavelengths (471 to 851 nm) and five spatial frequencies (0 to 0.2 mm−1). Three models were built from subsets of this initial dataset. The first subset included data taken at all wavelengths with the planar (0 mm−1) spatial frequency, the second comprised data at all wavelengths and spatial frequencies, and the third used all collected data at values relative to unburned tissue. These data subsets were used to train and test cubic SVM models, and compared against burn status 28 days after injury. Model accuracy was established through leave-one-out cross-validation testing. The model based on images obtained at all wavelengths and spatial frequencies predicted burn severity at 24 h with 92.5% accuracy. The model composed of all values relative to unburned skin was 94.4% accurate. By comparison, the model that employed only planar illumination was 88.8% accurate. This investigation suggests that the combination of SFDI with machine learning has potential for accurately predicting burn severity.

Cerebral oximetry based on near-infrared spectroscopy (NIRS) has seen increasing clinical use for monitoring of premature infants as well as during neonatal, pediatric and adult cardiac surgery. One key confounding factor and a likely contributor to observed inconsistency amongst commercial NIRS oximeters is skin pigmentation. Clinical studies have shown negative bias in oxygen saturation (StO2) with increasing melanin content. In prior work, we developed a cerebral oximetry phantom comprised of a 3D-printed channel array module representing brain tissue and molded silicone layers simulating extracranial regions. The purpose of the current study was to develop and test epidermis-simulating layers that exhibit realistic human pigmentation properties. Initially, we performed spectroscopic characterization of potential melanin simulating agents – including coffee, India ink, synthetic melanin, and water-soluble nigrosin – in a polydimethylsiloxane (PDMS) substrate. We determined that the NIR absorption spectrum of water-soluble nigrosin most accurately matched human melanin. Layers of 0.1 mm thickness were fabricated with different nigrosin concentrations to simulate epidermis with light, moderate, and dark pigmentation. The brain module channels were filled with bovine blood in the 30-100% oxygenation range and measurements performed with neonatal/pediatric probes from commercially available cerebral oximeters. We found that StO2 reported by the oximeters decreased monotonically with increasing pigmentation level. The magnitude of this impact increased with decreasing StO2, producing a maximum change in saturation of approximately 8%. The consistency of our results with prior clinical findings provides preliminary evidence of the utility of our approach for assessing the impact of epidermal melanin in phantom-based performance testing.

While most superficial (first degree) burns and full thickness (third degree) burns are easily diagnosed through standard clinical inspection, burns that fall between these extremes are challenging to accurately classify based on clinical appearance alone. Accurate early assessments have the potential to enable earlier debridement and grafting, thereby reducing complication rates, healing times, and chances for infection. To this end, several emerging technologies aim to objectively and non-invasively quantify burn severity, including Spatial Frequency Domain Imaging (SFDI), Laser Speckle Imaging (LSI), and infrared thermal imaging (IR). While these imaging modalities have some diagnostic ability for in-vivo assessment of burn severity, they have not been directly compared in a controlled experiment. Here we compare the ability of these techniques to assess the severity of graded burns in a porcine model (n=96 burns, 6 pigs). Biopsies were taken for histological analysis to verify burn severity. Clinical assessment, SFDI, LSI and thermal imaging were performed for all graded burns at 24 and 72 hours post-burn. In terms of accuracy of burn severity assessment, using histology as a reference, SFDI (85%) and clinical analysis (83%) performed significantly better that LSI (75%) and thermography (73%) at the 24 hour post-burn time point. There was no statistically significant improvement in assessment accuracy at the 72 hour post-burn time point, across the imaging modalities. These data indicate that SFDI may provide valuable information to clinicians in future clinical trials. This information can be obtained early in the process, which may hasten surgical intervention and reduce complications.

Assessment of burn severity is critical for wound treatment. Spatial frequency domain imaging (SFDI) has been previously used to characterize burns based on the relationships between histology and tissue optical properties. Recently, multispectral and hyperspectral imaging optical features have been combined with machine learning to classify burn severity. Here, we investigated the use of SFDI reflectance data at multiple wavelengths and spatial frequencies, with a support vector machine (SVM), to predict severity in a porcine model of graded burns. Burn severity predictions using SVM were compared to burn grade determined using histology techniques. Results suggest that the combination of spatial frequency data with machine learning models has the potential for accurately predicting burn severity at the 24 hr postburn time point.

Spatial frequency domain imaging (SFDI) is a wide-field imaging technique that enables quantification of in vivo tissue optical properties. SFDI was employed to evaluate burn wounds in pigs where incomplete debridement resulted in skin graft failure. We observed increase in the magnitude of the reduced-scattering coefficient in the wound beds after debridement, prior to grafting in wounds that failed compared to the reduced scattering measured in the wound beds that did not fail. We anticipate that SFDI may be a useful means for determining appropriate debridement prior to grafting.

Burn wounds and wound healing invoke several biological processes that may complicate the interpretation of spectral imaging data. Through analysis of spatial frequency domain spectroscopy data (450 to 1000nm) obtained from longitudinal investigations using a graded porcine burn wound healing model, we have identified features in the absorption spectrum that appear to suggest the presence of hemoglobin breakdown products, e.g., methemoglobin. Our results show that the calculated concentrations of methemoglobin directly correlate with burn severity, 24h after the injury. In addition, tissue parameters such as oxygenation (StO2) and water fraction may be underestimated by 20% and 78%, respectively, if methemoglobin is not included in the spectral analysis.

We introduce a method for quantitative hyperspectral optical imaging in the spatial frequency domain (hs-SFDI) to image tissue absorption (μa) and reduced scattering (μs') parameters over a broad spectral range. The hs-SFDI utilizes principles of spatial scanning of the spectrally dispersed output of a supercontinuum laser that is sinusoidally projected onto the tissue using a digital micromirror device. A scientific complementary metal-oxide-semiconductor camera is used for capturing images that are demodulated and analyzed using SFDI computational models. The hs-SFDI performance is validated using tissue-simulating phantoms over a range of μa and μs' values. Quantitative hs-SFDI images are obtained from an ex-vivo beef sample to spatially resolve concentrations of oxy-, deoxy-, and met-hemoglobin, as well as water and fat fractions. Our results demonstrate that the hs-SFDI can quantitatively image tissue optical properties with 1000 spectral bins in the 580- to 950-nm range over a wide, scalable field of view. With an average accuracy of 6.7% and 12.3% in μa and μs', respectively, compared to conventional methods, hs-SFDI offers a promising approach for quantitative hyperspectral tissue optical imaging. Funding agencies: National Institute of Health (NIH) NIBIB Biomedical Technology Research Center LAMMP [P41EB015890]; NIH [R21EB020953]; Military Medical Photonics Program: AFOSR [FA9550-17-1-0193]; Arnold and Mabel Beckman Foundation

We present a method to recover absorption and reduced scattering spectra for each layer of a two-layer turbid media from spatial frequency-domain spectroscopy data. We focus on systems in which the thickness of the top layer is less than the transport mean free path ( 0.1 − 0.8l * ) . We utilize an analytic forward solver, based upon the N’th-order spherical harmonic expansion with Fourier decomposition ( SHEFN ) method in conjunction with a multistage inverse solver. We test our method with data obtained using spatial frequency-domain spectroscopy with 32 evenly spaced wavelengths within λ = 450 to 1000 nm on six-layered tissue phantoms with distinct optical properties. We demonstrate that this approach can recover absorption and reduced scattering coefficient spectra for both layers with accuracy comparable with current Monte Carlo methods but with lower computational cost and potential flexibility to easily handle variations in parameters such as the scattering phase function or material refractive index. To our knowledge, this approach utilizes the most accurate deterministic forward solver used in such problems and can successfully recover properties from a two-layer media with superficial layer thicknesses.

The ability to predict success or failure of wound healing strategies in burns has the potential to enable customized treatments tailored for the individual patient that would ultimately shorten recovery times, reduce the need for repeated grafting, and aid in achieving full rehabilitation. To this end, we are investigating the potential for spatial frequency domain imaging (SFDI) to non-invasively assess wound severity and healing status. SFDI is a wide-field diffuse optical imaging technique that enables non-invasive quantitative determination of in vivo tissue optical properties associated with physiologic structure and function. We have employed SFDI in a longitudinal study of wound healing using a controlled porcine burn model. Regions of wound repair using split thickness skin grafts were imaged using SFDI at multiple time points over a period of 60 days. The reduced scattering and absorption coefficients were determined at nine wavelengths spanning 470–970 nm, enabling determination of oxy- and deoxy-hemoglobin concentration, total hemoglobin concentration, oxygen saturation, and water fraction. These values obtained were compared to unburned control regions, undebrided burns, and debrided regions without treatment by grafting. Changes in the reduced scattering associated with structural changes in tissue correlate with histology as the wound heals. Compared to unburned tissue, the reduced scattering coefficient associated with repaired wounds is depressed and is spatially heterogeneous immediately following grafting. Over the course of healing, the scattering values increase and converge toward values of normal tissue and become more spatially homogeneous. Variations in chromophore concentrations are also characterized.

Spatial frequency domain imaging (SFDI) utilizes a digital light processing (DLP) projector for illuminating turbid media with sinusoidal patterns. The tissue absorption (μ a ) and reduced scattering coefficient (μ , s ) are calculated by analyzing the modulation transfer function for at least two spatial frequencies. We evaluated different illumination strategies with a red, green and blue light emitting diodes (LED) in the DLP, while imaging with a filter mosaic camera, XiSpec, with 16 different multi-wavelength sensitive pixels in the 470-630 nm wavelength range. Data were compared to SFDI by a multispectral camera setup (MSI) consisting of four cameras with bandpass filters centered at 475, 560, 580 and 650 nm. A pointwise system for comprehensive microcirculation analysis was used (EPOS) for comparison. A 5-min arterial occlusion and release protocol on the forearm of a Caucasian male with fair skin was analyzed by fitting the absorption spectra of the chromophores HbO 2 , Hb and melanin to the estimatedμ a . The tissue fractions of red blood cells ( f RBC), melanin (/mel) and the Hb oxygenation ( S 0 2 ) were calculated at baseline, end of occlusion, early after release and late after release. EPOS results showed a decrease in S 0 2 during the occlusion and hyperemia during release ( S 0 2 = 40%, 5%, 80% and 51%). The f RBC showed an increase during occlusion and release phases. The best MSI resemblance to the EPOS was for green LED illumination ( S 0 2 = 53%, 9%, 82%, 65%). Several illumination and analysis strategies using the XiSpec gave un-physiological results (e.g. negative S 0 2 ). XiSpec with green LED illumination gave the expected change in /RBC , while the dynamics in S 0 2 were less than those for EPOS. These results may be explained by the calculation of modulation using an illumination and detector setup with a broad spectral transmission bandwidth, with considerable variation in μ a of included chromophores. Approaches for either reducing the effective bandwidth of the XiSpec filters or by including their characteristic in a light transport model for SFDI modulation, are proposed.

Hyperspectral Imaging (HSI) is a growing field in tissue optics due to its ability to collect continuous spectral features of a sample without a contact probe. Spatial Frequency Domain Imaging (SFDI) is a non-contact wide-field spectral imaging technique that is used to quantitatively characterize tissue structure and chromophore concentration. In this study, we designed a Hyperspectral SFDI (H-SFDI) instrument which integrated a supercontinuum laser source to a wavelength tuning optical configuration and a sCMOS camera to extract spatial (Field of View: 2cm×2cm) and broadband spectral features (580nm-950nm). A preliminary experiment was also performed to integrate the hyperspectral projection unit to a compressed single pixel camera and Light Labeling (LiLa) technique.

Author(s): Ponticorvo, Adrien; Taydas, Eren; Mazhar, Amaan; Ellstrom, Christopher L; Rimler, Jonathan; Scholz, Thomas; Tong, June; Evans, Gregory R D; Cuccia, David J; Durkin, Anthony J | Abstract: Detecting failing tissue flaps before they are clinically apparent has the potential to improve postoperative flap management and salvage rates. This study demonstrates a model to quantitatively compare clinical appearance, as recorded via digital camera, with spatial frequency domain imaging (SFDI), a noninvasive imaging technique using patterned illumination to generate images of total hemoglobin and tissue oxygen saturation (stO2).Using a swine pedicle model in which blood flow was carefully controlled with occlusion cuffs and monitored with ultrasound probes, throughput was reduced by 25%, 50%, 75%, and 100% of baseline values in either the artery or the vein of each of the flaps. The color changes recorded by a digital camera were quantified to predict which occlusion levels were visible to the human eye. SFDI was also used to quantify the changes in physiological parameters including total hemoglobin and oxygen saturation associated with each occlusion.There were no statistically significant changes in color above the noticeable perception levels associated with human vision during any of the occlusion levels. However, there were statistically significant changes in total hemoglobin and stO2 levels detected at the 50%, 75%, and 100% occlusion levels for arterial and venous occlusions.As demonstrated by the color imaging data, visual flap changes are difficult to detect until significant occlusion has occurred. SFDI is capable of detecting changes in total hemoglobin and stO2 as a result of partial occlusions before they are perceivable, thereby potentially improving response times and salvage rates.

With recent proliferation in compact and/or low-cost clinical multispectral imaging approaches and commercially available components, questions remain whether they adequately capture the requisite spectral content of their applications. We present a method to emulate the spectral range and resolution of a variety of multispectral imagers, based on in-vivo data acquired from spatial frequency domain spectroscopy (SFDS). This approach simulates spectral responses over 400 to 1100 nm. Comparing emulated data with full SFDS spectra of in-vivo tissue affords the opportunity to evaluate whether the sparse spectral content of these imagers can (1) account for all sources of optical contrast present (completeness) and (2) robustly separate and quantify sources of optical contrast (crosstalk). We validate the approach over a range of tissue-simulating phantoms, comparing the SFDS-based emulated spectra against measurements from an independently characterized multispectral imager. Emulated results match the imager across all phantoms ([Formula: see text] absorption, [Formula: see text] reduced scattering). In-vivo test cases (burn wounds and photoaging) illustrate how SFDS can be used to evaluate different multispectral imagers. This approach provides an in-vivo measurement method to evaluate the performance of multispectral imagers specific to their targeted clinical applications and can assist in the design and optimization of new spectral imaging devices.

© 2017 Author(s). Spatial Frequency Domain Spectroscopy (SFDS) is a technique for quantifying in-vivo tissue optical properties. SFDS employs structured light patterns that are projected onto tissues using a spatial light modulator, such as a digital micromirror device. In combination with appropriate models of light propagation, this technique can be used to quantify tissue optical properties (absorption, μa, and scattering, μs′, coefficients) and chromophore concentrations. Here we present a handheld implementation of an SFDS device that employs line (one dimensional) imaging. This instrument can measure 1088 spatial locations that span a 3 cm line as opposed to our original benchtop SFDS system that only collects a single 1 mm diameter spot. This imager, however, retains the spectral resolution (∼1 nm) and range (450-1000 nm) of our original benchtop SFDS device. In the context of homogeneous turbid media, we demonstrate that this new system matches the spectral response of our original system to within 1% across a typical range of spatial frequencies (0-0.35 mm-1). With the new form factor, the device has tremendously improved mobility and portability, allowing for greater ease of use in a clinical setting. A smaller size also enables access to different tissue locations, which increases the flexibility of the device. The design of this portable system not only enables SFDS to be used in clinical settings but also enables visualization of properties of layered tissues such as skin.

Tissue simulating phantoms provide a valuable platform for quantitative evaluation of the performance of diffuse optical devices. In this paper we report the development of a poly(dimethylsiloxane) (PDMS) tissue phantom that mimics the spectral characteristics of tissue water. We have developed these phantoms to mimic different water fractions in tissue for testing new devices within the context of clinical applications such as burn wound triage. Compared to liquid phantoms, PDMS phantoms are easier to transport and use, and have a longer usable life than gelatin based phantoms. The pthalocyanine dye 9606 was used to provide an absorption feature of in the vicinity of 970 nm. Scattering properties were independently determined by adding titanium dioxide powder to obtain reduced scattering coefficients similar to that of tissue in the near infrared. Phantom properties were characterized using the techniques of inverse adding doubling and spatial frequency domain imaging. Results presented here demonstrate that we can fabricate solid phantoms that can be used to simulate different water fractions.