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1.  Design and evaluation of a laboratory prototype system for 3D photoacoustic full breast tomography 
Biomedical Optics Express  2013;4(11):2555-2569.
Photoacoustic imaging can visualize vascularization-driven optical absorption contrast with great potential for breast cancer detection and diagnosis. State-of-the-art photoacoustic breast imaging systems are promising but are limited either by only a 2D imaging capability or by an insufficient imaging field-of-view (FOV). We present a laboratory prototype system designed for 3D photoacoustic full breast tomography, and comprehensively characterize it and evaluate its performance in imaging phantoms. The heart of the system is an ultrasound detector array specifically developed for breast imaging and optimized for high sensitivity. Each detector element has an acoustic lens to enlarge the acceptance angle of the large surface area detector elements to ensure a wide system FOV. We characterized the ultrasound detector array performance in terms of frequency response, directional sensitivity, minimum detectable pressure and inter-element electrical and mechanical cross-talk. Further we evaluated the system performance of the laboratory prototype imager using well-defined breast mimicking phantoms. The system possesses a 2 mm XY plane resolution and a 6 mm vertical resolution. A vasculature mimicking object was successfully visualized down to a depth of 40 mm in the breast phantom. Further, tumor mimicking spherical objects with 5 and 10 mm diameter at 20 mm and 40 mm depths are recovered, indicating high system sensitivity. The system has a 170 × 170 × 170 mm3 FOV, which is well suited for full breast imaging. Various recommendations are provided for performance improvement and to guide this laboratory prototype to a clinical version in future.
doi:10.1364/BOE.4.002555
PMCID: PMC3829550  PMID: 24298416
(110.5120) Photoacoustic imaging; (040.0040) Detectors; (170.3830) Mammography; (110.6955) Tomographic imaging
2.  Quantitative comparison of analysis methods for spectroscopic optical coherence tomography 
Biomedical Optics Express  2013;4(11):2570-2584.
Spectroscopic optical coherence tomography (sOCT) enables the mapping of chromophore concentrations and image contrast enhancement in tissue. Acquisition of depth resolved spectra by sOCT requires analysis methods with optimal spectral/spatial resolution and spectral recovery. In this article, we quantitatively compare the available methods, i.e. the short time Fourier transform (STFT), wavelet transforms, the Wigner-Ville distribution and the dual window method through simulations in tissue-like media. We conclude that all methods suffer from the trade-off in spectral/spatial resolution, and that the STFT is the optimal method for the specific application of the localized quantification of hemoglobin concentration and oxygen saturation.
doi:10.1364/BOE.4.002570
PMCID: PMC3829551  PMID: 24298417
(030.1640) Coherence; (070.4790) Spectrum analysis; (160.4760) Optical properties; (170.6510) Spectroscopy, tissue diagnostics
3.  A literature review and novel theoretical approach on the optical properties of whole blood 
Lasers in Medical Science  2013;29:453-479.
Optical property measurements on blood are influenced by a large variety of factors of both physical and methodological origin. The aim of this review is to list these factors of influence and to provide the reader with optical property spectra (250–2,500 nm) for whole blood that can be used in the practice of biomedical optics (tabulated in the appendix). Hereto, we perform a critical examination and selection of the available optical property spectra of blood in literature, from which we compile average spectra for the absorption coefficient (μa), scattering coefficient (μs) and scattering anisotropy (g). From this, we calculate the reduced scattering coefficient (μs′) and the effective attenuation coefficient (μeff). In the compilation of μa and μs, we incorporate the influences of absorption flattening and dependent scattering (i.e. spatial correlations between positions of red blood cells), respectively. For the influence of dependent scattering on μs, we present a novel, theoretically derived formula that can be used for practical rescaling of μs to other haematocrits. Since the measurement of the scattering properties of blood has been proven to be challenging, we apply an alternative, theoretical approach to calculate spectra for μs and g. Hereto, we combine Kramers–Kronig analysis with analytical scattering theory, extended with Percus–Yevick structure factors that take into account the effect of dependent scattering in whole blood. We argue that our calculated spectra may provide a better estimation for μs and g (and hence μs′ and μeff) than the compiled spectra from literature for wavelengths between 300 and 600 nm.
doi:10.1007/s10103-013-1446-7
PMCID: PMC3953607  PMID: 24122065
Blood; Optical properties; Spectroscopy; Absorption coefficient; Scattering coefficient; Scattering anisotropy
4.  Quantitative laser speckle flowmetry of the in vivo microcirculation using sidestream dark field microscopy 
Biomedical Optics Express  2013;4(11):2347-2361.
We present integrated Laser Speckle Contrast Imaging (LSCI) and Sidestream Dark Field (SDF) flowmetry to provide real-time, non-invasive and quantitative measurements of speckle decorrelation times related to microcirculatory flow. Using a multi exposure acquisition scheme, precise speckle decorrelation times were obtained. Applying SDF-LSCI in vitro and in vivo allows direct comparison between speckle contrast decorrelation and flow velocities, while imaging the phantom and microcirculation architecture. This resulted in a novel analysis approach that distinguishes decorrelation due to flow from other additive decorrelation sources.
doi:10.1364/BOE.4.002347
PMCID: PMC3829532  PMID: 24298399
(170.2655) Functional monitoring and imaging; (170.3880) Medical and biological imaging; (170.6480) Spectroscopy, speckle; (170.0180) Microscopy
5.  Spectral domain detection in low-coherence spectroscopy 
Biomedical Optics Express  2012;3(9):2263-2272.
Low-coherence spectroscopy (LCS) offers the valuable possibility to measure quantitative and wavelength resolved optical property spectra within a tissue volume of choice that is controllable both in size and in depth. Until now, only time domain detection was investigated for LCS (tdLCS), but spectral domain detection offers a theoretical speed/sensitivity advantage over tdLCS. In this article, we introduce a method for spectral domain detection in LCS (sdLCS), with optimal sensitivity as a function of measurement depth. We validate our method computationally in a simulation and experimentally on a phantom with known optical properties. The attenuation, absorption and scattering coefficient spectra from the phantom that were measured by sdLCS agree well with the expected optical properties and the measured optical properties by tdLCS.
doi:10.1364/BOE.3.002263
PMCID: PMC3447566  PMID: 23024918
(030.1640) Coherence; (300.6190) Spectrometers; (160.4760) Optical properties; (170.6510) Spectroscopy, tissue diagnostics
6.  How the blood pool properties at onset affect the temporal behavior of simulated bruises 
The influence of initial blood pool properties on the temporal behavior of bruises is currently unknown. We addressed this important issue by utilizing three typical classes of bruises in our three-layered finite compartment model. We simulated the effects of their initial shapes, regularity of boundaries and initial blood concentration distributions (gaussian vs. homogeneous) on the hemoglobin and bilirubin areas in the dermal top layer. Age determination of bruises with gaussian hemoglobin concentration was also addressed. We found that the initial blood pool properties strongly affect bruise behavior. We determined the age of a 200-h simulated bruise with gaussian hemoglobin concentration with 3 h uncertainty. In conclusion, bruise behavior depends non-intuitively on the initial blood pool properties; hence, a model that includes shape, area and concentration distribution at onset is indispensable. Future age determination, including inhomogeneous hemoglobin distributions, will likely be based on the presented method for gaussian distributions.
doi:10.1007/s11517-012-0860-5
PMCID: PMC3272227  PMID: 22261914
Bruise; Numerical modeling; Age determination
7.  Biphasic Oxidation of Oxy-Hemoglobin in Bloodstains 
PLoS ONE  2011;6(7):e21845.
Background
In forensic science, age determination of bloodstains can be crucial in reconstructing crimes. Upon exiting the body, bloodstains transit from bright red to dark brown, which is attributed to oxidation of oxy-hemoglobin (HbO2) to met-hemoglobin (met-Hb) and hemichrome (HC). The fractions of HbO2, met-Hb and HC in a bloodstain can be used for age determination of bloodstains. In this study, we further analyze the conversion of HbO2 to met-Hb and HC, and determine the effect of temperature and humidity on the conversion rates.
Methodology
The fractions of HbO2, met-Hb and HC in a bloodstain, as determined by quantitative analysis of optical reflectance spectra (450–800 nm), were measured as function of age, temperature and humidity. Additionally, Optical Coherence Tomography around 1300 nm was used to confirm quantitative spectral analysis approach.
Conclusions
The oxidation rate of HbO2 in bloodstains is biphasic. At first, the oxidation of HbO2 is rapid, but slows down after a few hours. These oxidation rates are strongly temperature dependent. However, the oxidation of HbO2 seems to be independent of humidity, whereas the transition of met-Hb into HC strongly depends on humidity. Knowledge of these decay rates is indispensable for translating laboratory results into forensic practice, and to enable bloodstain age determination on the crime scene.
doi:10.1371/journal.pone.0021845
PMCID: PMC3137592  PMID: 21789186
8.  Optical mammography combined with fluorescence imaging: lesion detection using scatterplots 
Biomedical Optics Express  2011;2(4):1007-1020.
Using scatterplots of 2 or 3 parameters, diffuse optical tomography and fluorescence imaging are combined to improve detectability of breast lesions. Small or low contrast phantom-lesions that were missed in the optical and fluorescence images were detected in the scatterplots. In patient measurements, all tumors were visible and easily differentiated from artifacts and areolas in the scatterplots. The different rate of intake and wash out of the fluorescent contrast agent in the healthy versus malignant tissues was also observed in the scatterplot: this information can be used to discriminate malignant lesion from normal structures.
doi:10.1364/BOE.2.001007
PMCID: PMC3072114  PMID: 21483622
(170.3880) Medical and biological imaging; (170.6280) Spectroscopy, fluorescence, luminescence; (170.6960) Tomography; (170.3830) Mammography
9.  Non-contact spectroscopic determination of large blood volume fractions in turbid media 
Biomedical Optics Express  2011;2(2):396-407.
We report on a non-contact method to quantitatively determine blood volume fractions in turbid media by reflectance spectroscopy in the VIS/NIR spectral wavelength range. This method will be used for spectral analysis of tissue with large absorption coefficients and assist in age determination of bruises and bloodstains. First, a phantom set was constructed to determine the effective photon path length as a function of μa and μs′ on phantoms with an albedo range: 0.02-0.99. Based on these measurements, an empirical model of the path length was established for phantoms with an albedo > 0.1. Next, this model was validated on whole blood mimicking phantoms, to determine the blood volume fractions ρ = 0.12-0.84 within the phantoms (r = 0.993; error < 10%). Finally, the model was proved applicable on cotton fabric phantoms.
doi:10.1364/BOE.2.000396
PMCID: PMC3038454  PMID: 21339884
(170.1470) Blood or tissue constituent monitoring; (300.1030) Spectroscopy; (000.1430) Biology and medicine
10.  3D finite compartment modeling of formation and healing of bruises may identify methods for age determination of bruises 
Simulating the spatial and temporal behavior of bruises may identify methods that allow accurate age determination of bruises to assess child abuse. We developed a numerical 3D model to simulate the spatial kinetics of hemoglobin and bilirubin during the formation and healing of bruises. Using this model, we studied how skin thickness, bruise diameter and diffusivities affect the formation and healing of circular symmetric bruises and compared a simulated bruise with a natural inhomogeneous bruise. Healing is faster for smaller bruises in thinner and less dense skin. The simulated and natural bruises showed similar spatial and temporal dynamics. The different spatio-temporal dynamics of hemoglobin and bilirubin allows age determination of model bruises. Combining our model predictions with individual natural bruises may allow optimizing our model parameters. It may particularly identify methods for more accurate age determination than currently possible to aid the assessment of child abuse.
doi:10.1007/s11517-010-0647-5
PMCID: PMC2926474  PMID: 20556661
Bruise; Numerical modeling; Age determination; Child abuse; Diffusion
11.  Apoptosis- and necrosis-induced changes in light attenuation measured by optical coherence tomography 
Lasers in Medical Science  2009;25(2):259-267.
Optical coherence tomography (OCT) was used to determine optical properties of pelleted human fibroblasts in which necrosis or apoptosis had been induced. We analysed the OCT data, including both the scattering properties of the medium and the axial point spread function of the OCT system. The optical attenuation coefficient in necrotic cells decreased from 2.2 ± 0.3 mm−1 to 1.3 ± 0.6 mm−1, whereas, in the apoptotic cells, an increase to 6.4 ± 1.7 mm−1 was observed. The results from cultured cells, as presented in this study, indicate the ability of OCT to detect and differentiate between viable, apoptotic, and necrotic cells, based on their attenuation coefficient. This functional supplement to high-resolution OCT imaging can be of great clinical benefit, enabling on-line monitoring of tissues, e.g. for feedback in cancer treatment.
doi:10.1007/s10103-009-0723-y
PMCID: PMC2807596  PMID: 19756838
Optical coherence tomography; Optical properties; Cells; Apoptosis; Necrosis
12.  Combined Raman spectroscopy and optical coherence tomography device for tissue characterization 
Optics letters  2008;33(10):1135-1137.
We report a dual-modal device capable of sequential acquisition of Raman spectroscopy (RS) and optical coherence tomography (OCT) along a common optical axis. The device enhances application of both RS and OCT by precisely guiding RS acquisition with OCT images while also compensating for the lack of molecular specificity in OCT with the biochemical specificity of RS. We characterize the system performance and demonstrate the capability to identify structurally ambiguous features within an OCT image with RS in a scattering phantom, guide acquisition of RS from a localized malignancy in ex vivo breast tissue, and perform in vivo tissue analysis of a scab.
PMCID: PMC2713918  PMID: 18483537
13.  Synthesis and Bioconjugation of Gold Nanoparticles as Potential Molecular Probes for Light-Based Imaging Techniques 
We have synthesized and characterized gold nanoparticles (spheres and rods) with optical extinction bands within the “optical imaging window.” The intense plasmon resonant driven absorption and scattering peaks of these nanoparticles make them suitable as contrast agents for optical imaging techniques. Further, we have conjugated these gold nanoparticles to a mouse monoclonal antibody specific to HER2 overexpressing SKBR3 breast carcinoma cells. The bioconjugation protocol uses noncovalent modes of binding based on a combination of electrostatic and hydrophobic interactions of the antibody and the gold surface. We discuss various aspects of the synthesis and bioconjugation protocols and the characterization results of the functionalized nanoparticles. Some proposed applications of these potential molecular probes in the field of biomedical imaging are also discussed.
doi:10.1155/2007/29817
PMCID: PMC2266791  PMID: 18354723
14.  A Clinical Instrument for Combined Raman Spectroscopy-Optical Coherence Tomography of Skin Cancers 
Lasers in surgery and medicine  2011;43(2):143-151.
Background and Objective
The current standard for diagnosis of skin cancers is visual inspection followed by biopsy and histopathology. This process can be invasive, subjective, time consuming, and costly. Optical techniques, including Optical Coherence Tomography (OCT) and Raman Spectroscopy (RS), have been developed to perform non-invasive characterization of skin lesions based on either morphological or biochemical features of disease. The objective of this work is to report a clinical instrument capable of both morphological and biochemical characterization of skin cancers with RS-OCT.
Materials and Methods
The portable instrument utilizes independent 785 nm RS and 1310 nm OCT system backbones. The two modalities are integrated in a 4” (H) × 5”(W) × 8”(L) clinical probe. The probe enables sequential acquisition of co-registered OCT and RS data sets. The axial response of the RS collection in the skin was estimated using scattering phantoms. In addition, RS-OCT data from patients with cancerous and non-cancerous lesions are reported.
Results
The RS-OCT instrument is capable of screening areas as large as 15 mm (transverse) by 2.4 mm (in depth) at up to 8 frames/sec with OCT, and identifying locations to perform RS. RS signal is collected from a 44 µm transverse spot through a depth of approximately 530 µm. RS-OCT data sets from a superficial scar and a nodular BCC are reported to demonstrate the clinical potential of the instrument.
Conclusion
The RS-OCT instrument reported here is capable of morphological and biochemical characterization of cancerous skin lesions in a clinical setting. OCT can visualize microstructural irregularities and perform an initial morphological analysis of the lesion. The images can be used to guide acquisition of biochemically specific Raman spectra. The two data sets can then be evaluated with respect to one another to take advantage of the mutually complimentary nature of RS and OCT.
doi:10.1002/lsm.21041
PMCID: PMC4014065  PMID: 21384396

Results 1-14 (14)