Renal cortex microperfusion evaluated by laser speckle contrast imaging in an ex vivo perfused kidney model-A proof-of-concept study.

BACKGROUND: Validated quantitative biomarkers for assessment of renal graft function during normothermic machine perfusion (NMP) conditions are lacking. The aim of this project was to quantify cortex microperfusion during ex vivo kidney perfusion using laser speckle contrast imaging (LSCI), and to evaluate the sensitivity of LSCI when measuring different levels of renal perfusion. Furthermore, we aimed to introduce LSCI measurements during NMP in differentially damaged kidneys. METHODS: Eleven porcine kidneys were nephrectomized and perfused ex vivo. Cortex microperfusion was simultaneously monitored using LSCI. First, a flow experiment examined the relationship between changes in delivered renal flow and corresponding changes in LSCI-derived cortex microperfusion. Second, renal cortical perfusion was reduced stepwise by introducing a microembolization model. Finally, LSCI was applied for measuring renal cortex microperfusion in kidneys exposed to minimal damage or 2 h warm ischemia (WI). RESULTS: Cortex microperfusion was calculated from the LSCI-obtained data. The flow experiment resulted in relatively minor changes in cortex microperfusion compared to the pump-induced changes in total renal flow. Based on stepwise injections of microspheres, we observed different levels of cortex microperfusion that correlated with administrated microsphere dosages (r2 = 0.95-0.99). We found no difference in LSCI measured cortex microperfusion between the kidneys exposed to minimal damage (renal cortex blood flow index, rcBFI = 2090-2600) and 2 h WI (rcBFI = 2189-2540). CONCLUSIONS: Based on this preliminary study, we demonstrated the feasibility of LSCI in quantifying cortex microperfusion during ex vivo perfusion. Furthermore, based on LSCI-measurements, cortical microperfusion was similar in kidneys exposed to minimal and 2 h WI.

Glutathione Protects the Developing Heart from Defects and Global DNA Hypomethylation Induced by Prenatal Alcohol Exposure.

BACKGROUND: Fetal alcohol spectrum disorder (FASD) is caused by prenatal alcohol exposure (PAE), the intake of ethanol (C2 H5 OH) during pregnancy. Features of FASD cover a range of structural and functional defects including congenital heart defects (CHDs). Folic acid and choline, contributors of methyl groups to one-carbon metabolism (OCM), prevent CHDs in humans. Using our avian model of FASD, we have previously reported that betaine, another methyl donor downstream of choline, prevents CHDs. The CHD preventions are substantial but incomplete. Ethanol causes oxidative stress as well as depleting methyl groups for OCM to support DNA methylation and other epigenetic alterations. To identify more compounds that can safely and effectively prevent CHDs and other effects of PAE, we tested glutathione (GSH), a compound that regulates OCM and is known as a "master antioxidant." METHODS/RESULTS: Quail embryos injected with a single dose of ethanol at gastrulation exhibited congenital defects including CHDs similar to those identified in FASD individuals. GSH injected simultaneously with ethanol not only prevented CHDs, but also improved survival and prevented other PAE-induced defects. Assays of hearts at 8 days (HH stage 34) of quail development, when the heart normally has developed 4-chambers, showed that this single dose of PAE reduced global DNA methylation. GSH supplementation concurrent with PAE normalized global DNA methylation levels. The same assays performed on quail hearts at 3 days (HH stage 19-20) of development, showed no difference in global DNA methylation between controls, ethanol-treated, GSH alone, and GSH plus ethanol-treated cohorts. CONCLUSIONS: GSH supplementation shows promise to inhibit effects of PAE by improving survival, reducing the incidence of morphological defects including CHDs, and preventing global hypomethylation of DNA in heart tissues.

Complex regression Doppler optical coherence tomography.

We introduce a new method to measure Doppler shifts more accurately and extend the dynamic range of Doppler optical coherence tomography (OCT). The two-point estimate of the conventional Doppler method is replaced with a regression that is applied to high-density B-scans in polar coordinates. We built a high-speed OCT system using a 1.68-MHz Fourier domain mode locked laser to acquire high-density B-scans (16,000 A-lines) at high enough frame rates (∼100 fps) to accurately capture the dynamics of the beating embryonic heart. Flow phantom experiments confirm that the complex regression lowers the minimum detectable velocity from 12.25 mm / s to 374 µm / s, whereas the maximum velocity of 400 mm / s is measured without phase wrapping. Complex regression Doppler OCT also demonstrates higher accuracy and precision compared with the conventional method, particularly when signal-to-noise ratio is low. The extended dynamic range allows monitoring of blood flow over several stages of development in embryos without adjusting the imaging parameters. In addition, applying complex averaging recovers hidden features in structural images.

Complex decorrelation averaging in optical coherence tomography: a way to reduce the effect of multiple scattering and improve image contrast in a dynamic scattering medium.

We demonstrate that complex decorrelation averaging can reduce the effect of multiple scattering and improve optical coherence tomography (OCT) imaging contrast. Complex decorrelation averaging calculates the product of an A-scan and the complex conjugate of a subsequent A-scan. The resultant signal is the product of the amplitudes and the phase difference. All these resulting complex signals at a particular location are then averaged. We take advantage of the fact that complex averaging, in contrast to conventional magnitude averaging, is sensitive to phase decorrelation. Sample motion that increases signal phase variance results in lower signal magnitude after complex averaging. Such motion preferentially results in a faster decorrelation of the multiple scattering signal when compared to the single scattering signal with each scattering event spreading the phase. This indicates that we may reduce multiple scattering by implementing complex decorrelation averaging to preferentially reduce the magnitude of the multiply scattered light signal in OCT images. By adjusting the time between phase-differenced A-scans, one can regulate the amount of measured decorrelation. We have performed experiments on liquid phantoms that give experimental evidence for this hypothesis. A substantial improvement in OCT image contrast using complex decorrelation averaging is demonstrated.

Rotationally acquired four-dimensional optical coherence tomography of embryonic chick hearts using retrospective gating on the common central A-scan.

We introduce a new method of rotational image acquisition for four-dimensional (4D) optical coherence tomography (OCT) of beating embryonic chick hearts. The rotational axis and the central A-scan of the OCT are identical. An out-of-phase image sequence covering multiple heartbeats is acquired at every angle of an incremental rotation of the deflection mirrors of the OCT system. Image acquisition is accomplished after a rotation of 180°. Comparison of a displayed live M-mode of the central A-scan with a reference M-mode allows instant detection of translational movements of the embryo. For calculation of 4D data sets, we apply an image-based retrospective gating algorithm using the phase information of the common central A-scan present in all acquired images. This leads to cylindrical three-dimensional data sets for every time step of the cardiac cycle that can be used for 4D visualization. We demonstrate this approach and provide a video of a beating Hamburger and Hamilton stage 16 embryonic chick heart generated from a 4D OCT data set using rotational image acquisition.

Integration of an optical coherence tomography (OCT) system into an examination incubator to facilitate in vivo imaging of cardiovascular development in higher vertebrate embryos under stable physiological conditions.

High-resolution in vivo imaging of higher vertebrate embryos over short or long time periods under constant physiological conditions is a technically challenging task for researchers working on cardiovascular development. In chick embryos, for example, various studies have shown that without appropriate maintenance of temperature, as one of the main environmental factors, the embryonic heart rate drops rapidly and often results in an increase in regurgitant flow. Hemodynamic parameters are critical stimuli for cardiovascular development that, for a correct evaluation of their developmental significance, should be documented under physiological conditions. However, previous studies were mostly carried out outside of an incubator or under suboptimal environmental conditions. Here we present, to the best of our knowledge, the first detailed description of an optical coherence tomography (OCT) system integrated into an examination incubator to facilitate real-time in vivo imaging of cardiovascular development under physiological environmental conditions. We demonstrate the suitability of this OCT examination incubator unit for use in cardiovascular development studies by examples of proof of principle experiments. We, furthermore, point out the need for use of examination incubators for physiological OCT examinations by documenting the effects of room climate (22°C) on the performance of the cardiovascular system of chick embryos (HH-stages 16/17). Upon exposure to room climate, chick embryos showed a fast drop in the heart rate and striking changes in the cardiac contraction behaviour and the blood flow through the vitelline circulation. We have documented these changes for the first time by M-mode OCT and Doppler M-mode OCT.

How histological features of basal cell carcinomas influence image quality in optical coherence tomography.

Optical coherence tomography (OCT) has the potential to diagnose and measure the depth of nonmelanoma skin cancer (NMSC) in skin, but some lesions appear blurred in OCT images. The aim of this study is to identify histological characteristics of basal cell carcinomas (BCC) that correlate with good quality OCT images of the same lesions. A total of 34 patients with BCC were OCT scanned. The influence of histology parameters (e.g. inflammation, sun damage of skin, carcinoma cell size) on OCT image quality was studied by comparing 15 BCC lesions easily identified compared to 19 BCC lesions that produced only blurred in OCT images. Inflammation was more pronounced in blurred OCT images, whereas solar elastosis dominated in easily identified lesions. Hyperkeratosis did not impair imaging significantly. OCT image quality of BCC may depend on specific histology parameters.

Improved quality of optical coherence tomography imaging of basal cell carcinomas using speckle reduction.

BACKGROUND: Optical coherence tomography (OCT) is a possible imaging method for delineation of non-melanoma skin cancer. Speckle noise is the dominant noise contribution in OCT images; it limits the ability to identify cellular structures especially skin cancer. QUESTIONS ADDRESSED: This report suggests a method for improving OCT image quality for skin cancer imaging. EXPERIMENTAL DESIGN: OCT is an optical imaging method analogous to ultrasound. Two basal cell carcinomas (BCC) were imaged using an OCT speckle reduction technique (SR-OCT) based on repeated scanning by altering the distance between the probe and the surface of the skin. RESULTS: SR-OCT resulted in improved visualisation and more accurate thickness measurements in BCC lesions. CONCLUSION: This OCT speckle reduction method led to improved visualisation and better defined delineations in two BCC lesions. Thus, OCT was improved to a clinically relevant level when imaging BCC lesions.

Optical coherence tomography imaging of psoriasis vulgaris: correlation with histology and disease severity.

Epidermal thickness (ET) has been suggested as a surrogate measure of psoriasis severity. Optical coherence tomography (OCT) is a recent imaging technology that provides real-time skin images to a depth of 1.8 mm with a micrometre resolution. OCT may provide an accurate in vivo measure of ET. It is, therefore, speculated that OCT may be used in the assessment of psoriasis vulgaris. A total of 23 patients with psoriasis vulgaris were systematically evaluated by OCT imaging and skin biopsy during treatment. Biopsies were graded for disease severity, and additional evaluation was done by the physician via psoriasis area and severity index (PASI) score, and by the patient through measures such as self-administered PASI, psoriasis life stress inventory index and dermatology life quality index. ET was calculated from OCT images. In comparison to normal skin, psoriasis appeared with a more irregular surface with a stronger entrance signal, a serrated dermo-epidermal junction was found and a less signal intensity in the dermis as shown in OCT images. ET measured in untreated plaques was thicker reflecting epidermal hyperproliferation and inflammation. The changes were significantly correlated with the biopsy grading (r (2) = 0.41, p = 0.001) and ET significantly decreased with treatment (p = 0.0001). ET correlated significantly with self-reported measures of disease severity, but not with physician-assessed global PASI. The data suggest that OCT may be used to measure ET in psoriasis and the measurements correlate with several other parameters of disease severity. This implies that OCT assessment of psoriatic plaques may provide a useful method for non-invasive in vivo method to follow the evolution of psoriasis lesions.

In vivo imaging of the cyclic changes in cross-sectional shape of the ventricular segment of pulsating embryonic chick hearts at stages 14 to 17: a contribution to the understanding of the ontogenesis of cardiac pumping function.

The cardiac cycle-related deformations of tubular embryonic hearts were traditionally described as concentric narrowing and widening of a tube of circular cross-section. Using optical coherence tomography (OCT), we have recently shown that, during the cardiac cycle, only the myocardial tube undergoes concentric narrowing and widening while the endocardial tube undergoes eccentric narrowing and widening, having an elliptic cross-section at end-diastole and a slit-shaped cross-section at end-systole. Due to technical limitations, these analyses were confined to early stages of ventricular development (chick embryos, stages 10-13). Using a modified OCT-system, we now document, for the first time, the cyclic changes in cross-sectional shape of beating embryonic ventricles at stages 14 to 17. We show that during these stages (1) a large area of diminished cardiac jelly appears at the outer curvature of the ventricular region associated with formation of endocardial pouches; (2) the ventricular endocardial lumen acquires a bell-shaped cross-section at end-diastole and becomes compressed like a fireplace bellows during systole; (3) the contracting portions of the embryonic ventricles display stretching along its baso-apical axis at end-systole. The functional significance of our data is discussed with respect to early cardiac pumping function.

OCT imaging of skin cancer and other dermatological diseases.

Optical coherence tomography (OCT) provides clinicians and researchers with micrometer-resolution, in vivo, cross-sectional images of human skin up to several millimeter depth. This review of OCT imaging applied within dermatology covers the application of OCT to normal skin, and reports on a large number of applications in the fields of non-melanoma skin cancer, malignant melanomas, psoriasis and dermatitis, infestations, bullous skin diseases, tattoos, nails, haemangiomas, and other skin diseases.

Assessment of optical coherence tomography imaging in the diagnosis of non-melanoma skin cancer and benign lesions versus normal skin: observer-blinded evaluation by dermatologists and pathologists.

BACKGROUND: Optical coherence tomography (OCT) is an optical imaging technique that may be useful in diagnosis of non-melanoma skin cancer (NMSC). OBJECTIVES: To describe OCT features in NMSC such as actinic keratosis (AK) and basal cell carcinoma (BCC) and in benign lesions and to assess the diagnostic accuracy of OCT in differentiating NMSC from benign lesions and normal skin. METHODS AND MATERIALS: OCT and polarization-sensitive (PS) OCT from 104 patients were studied. Observer-blinded evaluation of OCT images from 64 BCCs, 1 baso-squamous carcinoma, 39 AKs, two malignant melanomas, nine benign lesions, and 105 OCT images from perilesional skin was performed; 50 OCT images of NMSC and 50 PS-OCT images of normal skin were evaluated twice. RESULTS: Sensitivity was 79% to 94% and specificity 85% to 96% in differentiating normal skin from lesions. Important features were absence of well-defined layering in OCT and PS-OCT images and dark lobules in BCC. Discrimination of AK from BCC had an error rate of 50% to 52%. CONCLUSION: OCT features in NMSC are identified, but AK and BCC cannot be differentiated. OCT diagnosis is less accurate than clinical diagnosis, but high accuracy in distinguishing lesions from normal skin, crucial for delineating tumor borders, was obtained.

Broadband light generation at approximately 1300 nm through spectrally recoiled solitons and dispersive waves.

We experimentally study the generation of broadband light at approximately 1300 nm from an 810 nm Ti:sapphire femtosecond pump laser. We use two photonic crystal fibers with a second infrared zero-dispersion wavelength (lambda Z2) and compare the efficiency of two schemes: in one fiber lambda Z2=1400 nm and the light at 1300 nm is composed of spectrally recoiled solitons; in the other fiber lambda Z2=1200 nm and the light at 1300 nm is composed of dispersive waves.

Morphology and epidermal thickness of normal skin imaged by optical coherence tomography.

BACKGROUND: Optical coherence tomography (OCT) is an optical imaging technology with a potential in the non-invasive diagnosis of skin cancer. To identify skin pathologies using OCT, it is of prime importance to establish baseline morphological features of normal skin. AIMS: The aim of this study is to describe normal skin morphology using OCT and polarization-sensitive OCT (PS-OCT), which is a way of representing birefringent tissue such as collagen in OCT images. Anatomical locations in 20 healthy volunteers were imaged, and epidermal thickness (ET) was measured and compared to age, gender and skin colour. METHODS: OCT imaging is based on infrared light reflection/backscatter from tissue. PS-OCT detects birefringence of tissue. Imaging was performed in 12 skin regions. ET was calculated from the OCT images. RESULTS: Normal skin has a layered structure. Layering is less pronounced in adults. In glabrous skin the stratum corneum is visible. Children had larger ET (p < 0.0001). Age had a negative correlation with ET (p < 0.05). No gender- or skin-type-related differences in ET were found. CONCLUSION: This study contributes to understanding OCT and PS-OCT images of normal skin and indicates that OCT can be used for both the qualitative and quantitative assessment of skin.

High-resolution in vivo imaging of the cross-sectional deformations of contracting embryonic heart loops using optical coherence tomography.

The embryonic heart tube consists of an outer myocardial tube, a middle layer of cardiac jelly, and an inner endocardial tube. It is said that tubular hearts pump the blood by peristaltoid contractions. The traditional concept of cardiac peristalsis sees the cyclic deformations of pulsating heart tubes as concentric narrowing and widening of tubes of circular cross-section. We have visualized the cross-sectional deformations of contracting embryonic hearts in chick embryos (HH-stages 9-17) using real-time high-resolution optical coherence tomography. Cardiac contractions are detected from HH-stage 10 onward. During the cardiac cycle, the myocardial tube undergoes concentric narrowing and widening while the endocardial tube undergoes eccentric narrowing and widening, having an elliptic cross-section at end-diastole and a slit-shaped cross-section at end-systole. The eccentric deformation of the endocardial tube is the consequence of an uneven distribution of the cardiac jelly. Our data show that the cyclic deformations of pulsating embryonic heart tubes run other than originally thought. There is evidence that heart tubes of elliptic cross-section might pump blood with a higher mechanical efficiency than those of circular-cross section. The uneven distribution of cardiac jelly seems to prefigure the future AV and cono-truncal endocardial cushions.

Imaging of intradermal tattoos by optical coherence tomography.

BACKGROUND/PURPOSE: Tattoos have become increasingly popular followed by a growing demand for tattoo removal, and yet there is little knowledge and monitoring of tattoo pigment deposition in skin layers. The purpose of this pilot study is to describe optical coherence tomography image characteristics of intradermal tattoos. METHODS: We included five black tattoos in 3 female volunteers, 39, 35 and 30 years old. In vivo imaging of tattoo pigments in the skin is possible with optical coherence tomography (OCT), a novel non-invasive, in vivo optical imaging technology with a resolution and a penetration in skin high enough for visualization of tattoo pigment in the dermis. RESULTS: In optical coherence tomography images tattoo pigments clusters appear as dark, homogenous vertical columns and structures in the papillary dermis. OCT-scanned normal skin (without tattoos) appeared to be free of this dark structure. CONCLUSION: We have demonstrated that OCT can be used to visualize clusters of light absorbing pigments in a predictable manner.

Imaging of cutaneous larva migrans by optical coherence tomography.

BACKGROUND: Cutaneous larva migrans is a parasitic skin eruption caused by migration of larvae of various nematodes. Diagnosis of cutaneous larva migrans is currently based on the clinical signs of the creeping eruption. We are investigating a new diagnostic technology called optical coherence tomography (OCT) , which is potentially able to visualize structures in the skin with an 8 microm resolution. This technology could therefore potentially allow rapid, non-invasive, in vivo diagnosis of infestations. METHOD: Clinical cases of cutaneous larva migrans (n=3) were studied. All patients had a characteristic itching, serpinginous eruption typical of cutaneous larva migrans. The parasites were acquired on beach holidays in Thailand and Malaysia. All skin lesions were imaged by an OCT system developed at Risoe National Laboratory, Denmark. RESULT: Two out of three patients showed a round to oval structure (diameter 0.3-0.5mm) in the epidermis, Thus distinct OCT morphology in skin areas affected by cutaneous larva migrans was demonstrated. The larvae were not visualized in any of the patients. CONCLUSION: This study demonstrates that OCT a novel optical imaging technology, can image the larva tunnel in the skin with adequate spatial resolution, but not the larvae itself. OCT has a potential in imaging of skin infestations.

Swept wavelength source in the 1 microm range.

We demonstrate scanning over 1.1 nm with a frequency shifting ring source using a Ytterbium doped fiber amplifier (YDFA). It is, to the best of our knowledge, the first time an YDFA has been used in this configuration, and operation in the 1-1.1 microm wavelength range is made possible. We demonstrate a novel timing scheme that suppresses unwanted Q-switching behavior. Finally, using a concatenated numerical amplifier model, we are able to accurately predict the behavior of the source.

Extraction of optical scattering parameters and attenuation compensation in optical coherence tomography images of multilayered tissue structures.

A recently developed analytical optical coherence tomography (OCT) model [Thrane et al., J. Opt. Soc. Am. A 17, 484 (2000)] allows the extraction of optical scattering parameters from OCT images, thereby permitting attenuation compensation in those images. By expanding this theoretical model, we have developed a new method for extracting optical scattering parameters from multilayered tissue structures in vivo. To verify this, we used a Monte Carlo (MC) OCT model as a numerical phantom to simulate the OCT signal for heterogeneous multilayered tissue. Excellent agreement between the extracted values of the optical scattering properties of the different layers and the corresponding input reference values of the MC simulation was obtained, which demonstrates the feasibility of the method for in vivo applications. This is to our knowledge the first time such verification has been obtained, and the results hold promise for expanding the functional imaging capabilities of OCT.

Advanced modelling of optical coherence tomography systems.

Analytical and numerical models for describing and understanding the light propagation in samples imaged by optical coherence tomography (OCT) systems are presented. An analytical model for calculating the OCT signal based on the extended Huygens-Fresnel principle valid both for the single and multiple scattering regimes is reviewed. An advanced Monte Carlo model for calculating the OCT signal is also reviewed, and the validity of this model is shown through a mathematical proof based on the extended Huygens-Fresnel principle. Moreover, for the first time the model is verified experimentally. From the analytical model, an algorithm for enhancing OCT images is developed: the so-called true-reflection algorithm in which the OCT signal may be corrected for the attenuation caused by scattering. For the first time, the algorithm is demonstrated by using the Monte Carlo model as a numerical tissue phantom. Such algorithm holds promise for improving OCT imagery and to extend the possibility for functional imaging.