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September/October 2009

Volume 14, Issue 5, Articles (05xxxx)

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Editorial

Select this Article OPEN ACCESS		Looking Back, Looking Ahead Bruce J. Tromberg J. Biomed. Opt. 14, 050101 (Nov 03, 2009); http://dx.doi.org/10.1117/1.3258205 Online Publication Date: Nov 03, 2009 Full Text: Read Online (HTML) \| Download PDF
		Abstract Unavailable

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JBO Letters

Select this Article OPEN ACCESS		Extended in vivo anterior eye-segment imaging with full-range complex spectral domain optical coherence tomography Johannes Jungwirth, Bernhard Baumann, Michael Pircher, Erich Götzinger, and Christoph K. Hitzenberger J. Biomed. Opt. 14, 050501 (Sep 08, 2009); http://dx.doi.org/10.1117/1.3213569 \| Cited 5 times Online Publication Date: Sep 08, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We demonstrate the capability of full-range complex (FRC) spectral domain optical coherence tomography (SD-OCT) to image the anterior eye segment from the cornea to the posterior surface of the lens. With an adapted spectrometer design, we developed a SD-OCT system with an extended normal (single half-space) depth range of 7 mm (in air). This OCT-intrinsic depth range was doubled with a FRC technique. We demonstrate the performance of our OCT system by imaging the whole anterior segment of a healthy human eye in vivo.

Select this Article OPEN ACCESS		On the possibility of time-lapse ultrahigh-resolution optical coherence tomography for bladder cancer grading Zhijia Yuan, Bai Chen, Hugang Ren, and Yingtian Pan J. Biomed. Opt. 14, 050502 (Sep 16, 2009); http://dx.doi.org/10.1117/1.3223246 \| Cited 8 times Online Publication Date: Sep 16, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract It has been recently demonstrated that the cellular details of bladder epithelium embedded in speckle noise can be uncovered with time-lapse ultrahigh-resolution optical coherence tomography (TL-uOCT) by proper time-lapse frame averaging that takes advantage of cellular micromotion in fresh biological tissue ex vivo. Here, spectral-domain 3-D TL-uOCT is reported to further improve the image fidelity, and new experimental evidence is presented to differentiate normal and cancerous nuclei of rodent bladder epithelia. Results of animal cancer study reveal that despite a slight overestimation (e.g., <10%) of nuclear size (D_N) to histological evaluation, TL-uOCT is capable of distinguishing normal (D_N ≈ 7 μm) and cancerous (e.g., high-grade D_N″ ≈ 13 μm) urothelia, which may potentially be very useful for enhancing the diagnosis of nonpapillary bladder cancer. More animal study is being conducted to examine the utility to differentiate hyperplasia, dysplasia, and carcinoma in situ.

Select this Article OPEN ACCESS		Optical investigation of diffusion of levofloxacin mesylate in agarose hydrogel Shuaixia Tan, Hongjun Dai, Juejie Wu, Ning Zhao, Xiaoli Zhang, and Jian Xu J. Biomed. Opt. 14, 050503 (Oct 01, 2009); http://dx.doi.org/10.1117/1.3227034 \| Cited 1 time Online Publication Date: Oct 01, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Real-time electronic speckle pattern interferometry method has been applied to study the diffusion behavior of levofloxacin mesylate (MSALVFX) in agarose hydrogel. The results show that the diffusivity of solute decreases with the increase of concentration of agarose and adapts to Kohlrausch’s law. Furthermore, Amsden’s model, based on the retardance effect associated with polymer chain flexibility, was employed to simulate the diffusion behavior. The consistent results suggest that the retardance effect dominates the diffusion process of MSALFVX in hydrogel; moreover, polymer chain flexibility greatly affects drug transport within the polymer matrix.

Select this Article OPEN ACCESS		Doppler optical microangiography improves the quantification of local fluid flow and shear stress within 3-D porous constructs Yali Jia, Lin An, and Ruikang K. Wang J. Biomed. Opt. 14, 050504 (Oct 07, 2009); http://dx.doi.org/10.1117/1.3233590 \| Cited 1 time Online Publication Date: Oct 07, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Traditional phase-resolved Doppler optical coherence tomography (DOCT) has been reported to have potential for characterizing local fluid flow within a microporous scaffold. In this work, we apply Doppler optical microangiography (DOMAG), a new imaging technique developed by combining optical microangiography (OMAG) with a phase-resolved method, for improved assessment of local fluid flow and its derived parameters, shear stress, and interconnectivity, within highly scattering porous constructs. Compared with DOCT, we demonstrate a dramatic improvement of DOMAG in quantifying flow-related properties within scaffolds in situ for functional tissue engineering.

Select this Article OPEN ACCESS		Simultaneous, noninvasive observation of elastic scattering, fluorescence and inelastic scattering as a monitor of blood flow and hematocrit in human fingertip capillary beds Joseph Chaiken, Jerry Goodisman, Bin Deng, Rebecca J. Bussjager, and George Shaheen J. Biomed. Opt. 14, 050505 (Oct 07, 2009); http://dx.doi.org/10.1117/1.3233629 \| Cited 5 times Online Publication Date: Oct 07, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We report simultaneous observation of elastic scattering, fluorescence, and inelastic scattering from in vivo near-infrared probing of human skin. Careful control of the mechanical force needed to obtain reliable registration of in vivo tissue with an appropriate optical system allows reproducible observation of blood flow in capillary beds of human volar side fingertips. The time dependence of the elastically scattered light is highly correlated with that of the combined fluorescence and Raman scattered light. We interpret this in terms of turbidity (the impeding effect of red blood cells on optical propagation to and from the scattering centers) and the changes in the volume percentages of the tissues in the irradiated volume with normal homeostatic processes. By fitting to a model, these measurements may be used to determine volume fractions of plasma and RBCs.

Select this Article OPEN ACCESS		Live imaging of rat embryos with Doppler swept-source optical coherence tomography Irina V. Larina, Kenryo Furushima, Mary E. Dickinson, Richard R. Behringer, and Kirill V. Larin J. Biomed. Opt. 14, 050506 (Oct 01, 2009); http://dx.doi.org/10.1117/1.3241044 \| Cited 3 times Online Publication Date: Oct 01, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The rat has long been considered an excellent system to study mammalian embryonic cardiovascular physiology, but has lacked the extensive genetic tools available in the mouse to be able to create single gene mutations. However, the recent establishment of rat embryonic stem cell lines facilitates the generation of new models in the rat embryo to link changes in physiology with altered gene function to define the underlying mechanisms behind congenital cardiovascular birth defects. Along with the ability to create new rat genotypes there is a strong need for tools to analyze phenotypes with high spatial and temporal resolution. Doppler OCT has been previously used for 3-D structural analysis and blood flow imaging in other model species. We use Doppler swept-source OCT for live imaging of early postimplantation rat embryos. Structural imaging is used for 3-D reconstruction of embryo morphology and dynamic imaging of the beating heart and vessels, while Doppler-mode imaging is used to visualize blood flow. We demonstrate that Doppler swept-source OCT can provide essential information about the dynamics of early rat embryos and serve as a basis for a wide range of studies on functional evaluation of rat embryo physiology.

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Research Papers

Select this Article		Effects of a topically applied wound ointment on epidermal wound healing studied by in vivo fluorescence laser scanning microscopy analysis Bernhard Lange-Asschenfeldt, Alena Alborova, Daniela Krüger-Corcoran, Alexa Patzelt, Heike Richter, Wolfram Sterry, Axel Kramer, Eggert Stockfleth, and Jürgen Lademann J. Biomed. Opt. 14, 054001 (Sep 08, 2009); http://dx.doi.org/10.1117/1.3213603 \| Cited 7 times Online Publication Date: Sep 08, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Epidermal wound healing is a complex and dynamic regenerative process necessary to reestablish skin integrity. Fluorescence confocal laser scanning microscopy (FLSM) is a noninvasive imaging technique that has previously been used for evaluation of inflammatory and neoplastic skin disorders in vivo and at high resolution. We employed FLSM to investigate the evolution of epidermal wound healing noninvasively over time and in vivo. Two suction blisters were induced on the volar forearms of the study participants, followed by removal of the epidermis. To study the impact of wound ointment on the process of reepithelization, test sites were divided into two groups, of which one test site was left untreated as a negative control. FLSM was used for serial/consecutive evaluations up to 8 days. FLSM was able to visualize the development of thin keratinocyte layers developing near the wound edge and around hair follicles until the entire epidermis has been reestablished. Wounds treated with the wound ointment were found to heal significantly faster than untreated wounds. This technique allows monitoring of the kinetics of wound healing noninvasively and over time, while offering new insights into the potential effects of topically applied drugs on the process of tissue repair.

Select this Article OPEN ACCESS		Virtual histology of the human heart using optical coherence tomography Christina M. Ambrosi, Nader Moazami, Andrew M. Rollins, and Igor R. Efimov J. Biomed. Opt. 14, 054002 (Sep 08, 2009); http://dx.doi.org/10.1117/1.3213604 \| Cited 4 times Online Publication Date: Sep 08, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Optical coherence tomography (OCT) allows for the visualization of micron-scale structures within nontransparent biological tissues. For the first time, we demonstrate the use of OCT in identifying components of the cardiac conduction system and other structures in the explanted human heart. Reconstructions of cardiac structures up to 2 mm below the tissue surface were achieved and validated with Masson Trichrome histology in atrial, ventricular, sinoatrial nodal, and atrioventricular nodal preparations. The high spatial resolution of OCT provides visualization of cardiac fibers within the myocardium, as well as elements of the cardiac conduction system; however, a limiting factor remains its depth penetration, demonstrated to be ∼ 2 mm in cardiac tissues. Despite its currently limited imaging depth, the use of OCT to identify the structural determinants of both normal and abnormal function in the intact human heart is critical in its development as a potential aid to intracardiac arrhythmia diagnosis and therapy.

Select this Article OPEN ACCESS		Multimodal, multiphoton microscopy and image correlation analysis for characterizing corneal thermal damage Wen Lo, Yu-Lin Chang, Jia-Shiu Liu, Chiu-Mei Hseuh, Vladimir Hovhannisyan, Shean-Jen Chen, Hsin-Yuan Tan, and Chen-Yuan Dong J. Biomed. Opt. 14, 054003 (Oct 01, 2009); http://dx.doi.org/10.1117/1.3213602 Online Publication Date: Oct 01, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We used the combination of multiphoton autofluorescence (MAF), forward second-harmonic generation (FWSHG), and backward second-harmonic generation (BWSHG) imaging for the qualitative and quantitative characterization of thermal damage of ex vivo bovine cornea. We attempt to characterize the structural alterations by qualitative MAF, FWSHG, and BWSHG imaging in the temperature range of 37 to 90°C. In addition to measuring the absolute changes in the three types of signals at the stromal surface, we also performed image correlation analysis between FWSHG and BWSHG and demonstrate that with increasing thermal damage, image correlation between FWSHG and BWSHG significantly increases. Our results show that while MAF and BWSHG intensities may be used as preliminary indicators of the extent of corneal thermal damage, the most sensitive measures are provided by the decay in FWSHG intensity and the convergence of FWSHG and BWSHG images.

Select this Article		Spatially sculpted laser scissors for study of DNA damage and repair Jared Stephens, Samarendra K. Mohanty, Suzanne Genc, Xiangduo Kong, Kyoko Yokomori, and Michael W. Berns J. Biomed. Opt. 14, 054004 (Sep 22, 2009); http://dx.doi.org/10.1117/1.3213601 \| Cited 1 time Online Publication Date: Sep 22, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We present a simple and efficient method for controlled linear induction of DNA damage in live cells. By passing a pulsed laser beam through a cylindrical lens prior to expansion, an elongated elliptical beam profile is created with the ability to expose controlled linear patterns while keeping the beam and the sample stationary. The length and orientation of the beam at the sample plane were reliably controlled by an adjustable aperture and rotation of the cylindrical lens, respectively. Localized immunostaining by the DNA double strand break (DSB) markers phosphorylated H2AX (γH2AX) and Nbs1 in the nuclei of HeLa cells exposed to the “line scissors” was shown via confocal imaging. The line scissors method proved more efficient than the scanning mirror and scanning stage methods at induction of DNA DSB damage with the added benefit of having a greater potential for high throughput applications.

Select this Article OPEN ACCESS		Cyanine-loaded lipid nanoparticles for improved in vivo fluorescence imaging Isabelle Texier, Mathieu Goutayer, Anabela Da Silva, Laurent Guyon, Nadia Djaker, Véronique Josserand, Emmanuelle Neumann, Jérôme Bibette, and Françoise Vinet J. Biomed. Opt. 14, 054005 (Sep 08, 2009); http://dx.doi.org/10.1117/1.3213606 \| Cited 14 times Online Publication Date: Sep 08, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Fluorescence is a very promising radioactive-free technique for functional imaging in small animals and, in the future, in humans. However, most commercial near-infrared dyes display poor optical properties, such as low fluorescence quantum yields and short fluorescence lifetimes. In this paper, we explore whether the encapsulation of infrared cyanine dyes within the core of lipid nanoparticles (LNPs) could improve their optical properties. Lipophilic dialkylcarbocyanines DiD and DiR are loaded very efficiently in 30–35-nm-diam lipid droplets stabilized in water by surfactants. No significant fluorescence autoquenching is observed up to 53 dyes per particle. Encapsulated in LNP, which are stable for more than one year at room temperature in HBS buffer (HEPES 0.02 M, EDTA 0.01 M, pH 5.5), DiD and DiR display far improved fluorescence quantum yields Φ (respectively, 0.38 and 0.25) and longer fluorescence lifetimes τ (respectively, 1.8 and 1.1 ns) in comparison to their hydrophilic counterparts Cy5 (Φ = 0.28, τ = 1.0 ns) and Cy7 (Φ = 0.13, τ = 0.57 ns). Moreover, dye-loaded LNPs are able to accumulate passively in various subcutaneous tumors in mice, thanks to the enhanced permeability and retention effect. These new fluorescent nanoparticles therefore appear as very promising labels for in vivo fluorescence imaging.

Select this Article OPEN ACCESS		Estimation of indocyanine green concentration in blood from fluorescence emission: application to hemodynamic assessment during hemodialysis Jean-Michel I. Maarek and Daniel P. Holschneider J. Biomed. Opt. 14, 054006 (Oct 12, 2009); http://dx.doi.org/10.1117/1.3233652 \| Cited 1 time Online Publication Date: Oct 12, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract There is considerable interest in assessing cardiovascular function noninvasively in patients receiving hemodialysis. A possible approach is to measure the blood concentration of bolus-injected indocyanine green dye and to apply the dye-dilution method for estimating cardiac output and blood volume. Blood ICG concentration can be derived from a measurement of the ICG fluorescence through the dialysis tubing if a simple and unique calibration relationship can be established between transmural fluorescence intensity and blood ICG concentration. We investigated this relationship using Monte Carlo simulations of light transport in blood with varying hematocrit and ICG concentrations and performed empiric measurements of optical absorption and ICG fluorescence emission to confirm our findings. The ICG fluorescence intensity measured at the blood surface, as well as the light intensity remitted by the blood, varied as hematocrit changes modified the absorption and scattering characteristics of the blood. Calibration relationships were developed between fluorescence intensity and ICG concentration that accounted for hematocrit changes. Combining the backreflected fluorescence and the reflected light measured near the point of illumination provided optimal signal intensity, linearity, and robustness to hematocrit changes. These results provide a basis for developing a noninvasive approach to derive optically circulating blood ICG concentration in hemodialysis circuits.

Select this Article		Photoacoustic tomography of small animal brain with a curved array transducer Xinmai Yang, Anastasios Maurudis, John Gamelin, Andres Aguirre, Quing Zhu, and Lihong V. Wang J. Biomed. Opt. 14, 054007 (Sep 16, 2009); http://dx.doi.org/10.1117/1.3227035 \| Cited 5 times Online Publication Date: Sep 16, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We present the application of a curved array photoacoustic tomographic imaging system that can provide rapid, high-resolution photoacoustic imaging of small animal brains. The system is optimized to produce a B-mode, 90-deg field-of-view image at sub-200-μm resolution at a frame rate of ∼ 1 frame/second when a 10-Hz pulse repetition rate laser is employed. By rotating samples, a complete 360-deg scan can be achieved within 15 s. In previous work, two-dimensional (2-D) ex vivo mouse brain cortex imaging has been reported. We report three-dimensional (3-D) small animal brain imaging obtained with the curved array system. The results are presented as a series of 2-D cross-sectional images. Besides structural imaging, the blood oxygen saturation of the animal brain cortex is also measured in vivo. In addition, the system can measure the time-resolved relative changes in blood oxygen saturation level in the small animal brain cortex. Last, ultrasonic gel coupling, instead of the previously adopted water coupling, is conveniently used in near-real-time 2-D imaging.

Select this Article OPEN ACCESS		Diagnosis of oral submucous fibrosis with optical coherence tomography Cheng-Kuang Lee, Meng-Tsan Tsai, Hsiang-Chieh Lee, Hsin-Ming Chen, Chun-Pin Chiang, Yih-Ming Wang, and C. C. Yang J. Biomed. Opt. 14, 054008 (Sep 22, 2009); http://dx.doi.org/10.1117/1.3233653 \| Cited 7 times Online Publication Date: Sep 22, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The epithelium (EP) thickness and the standard deviation (SD) of A-mode scan intensity in the laminar propria (LP) layer are used as effective indicators for the diagnosis of oral submucous fibrosis (OSF) based on the noninvasive clinical scanning of a swept-source optical coherence tomography (OCT) system of ∼6 μm in axial resolution (in tissue) and 103 dB in sensitivity. Compared with the corresponding parameters in healthy oral mucosal mucosa, in OSF mucosa, the EP thickness becomes smaller and the SD of A-mode scan intensity in the LP layer (LP SD) also becomes smaller. The LP SD can also be used for effectively differentiating OSF (small LP SD) from lesion (large LP SD). This application is particularly useful in the case of a lesion without a clear surface feature. Meanwhile, the use of the SD of A-mode scan intensity in the EP layer (EP SD) can further help in differentiating OSF (medium EP SD) from healthy oral mucosal (small EP SD) and lesion (large EP SD) conditions. Compared with the conventional method of maximum mouth opening measurement, the use of the proposed OCT scanning results can be a more effective technique for OSF diagnosis.

Select this Article		Characterization of an orange acceptor fluorescent protein for sensitized spectral fluorescence resonance energy transfer microscopy using a white-light laser Yuansheng Sun, Cynthia F. Booker, Sangeeta Kumari, Richard N. Day, Mike Davidson, and Ammasi Periasamy J. Biomed. Opt. 14, 054009 (Sep 16, 2009); http://dx.doi.org/10.1117/1.3227036 \| Cited 3 times Online Publication Date: Sep 16, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Orange fluorescent proteins (FPs) are attractive candidates as Förster resonance energy transfer (FRET) partners, bridging the gap between green and red/far-red FPs, but they pose significant challenges using common fixed laser wavelengths. We investigated monomeric Kusabira orange 2 (mKO2) FP as a FRET acceptor for monomeric teal FP (mTFP) as donor on a FRET standard construct using a fixed-distance amino acid linker, expressed in live cells. We quantified the apparent FRET efficiency (E%) of this construct, using sensitized spectral FRET microscopy on the Leica TCS SP5 X imaging system equipped with a white-light laser that allows choosing any excitation wavelength from 470 to 670 nm in 1-nm increments. The E% obtained in sensitized spectral FRET microscopy was then confirmed with fluorescence lifetime measurements. Our results demonstrate that mKO2 and mTFP are good FRET partners given proper imaging setups. mTFP was optimally excited by the Argon 458 laser line, and the 540-nm wavelength excitation for mKO2 was chosen from the white-light laser. The white-light laser generally extends the usage of orange and red/far-red FPs in sensitized FRET microscopy assays by tailoring excitation and emission precisely to the needs of the FRET pair.

Select this Article OPEN ACCESS		Vector correlation technique for pixel-wise detection of collagen fiber realignment during injurious tensile loading Kyle P. Quinn and Beth A. Winkelstein J. Biomed. Opt. 14, 054010 (Sep 16, 2009); http://dx.doi.org/10.1117/1.3227037 \| Cited 6 times Online Publication Date: Sep 16, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Excessive soft tissue loading can produce adverse structural and physiological changes in the absence of any visible tissue rupture. However, image-based analysis techniques to assess microstructural changes during loading without any visible rupture remain undeveloped. Quantitative polarized light imaging (QPLI) can generate spatial maps of collagen fiber alignment during loading with high temporal resolution and can provide a useful technique to measure microstructural responses. While collagen fibers normally realign in the direction that tissue is loaded, rapid, atypical fiber realignment during loading may be associated with the response of a local collagenous network to fiber failure. A vector correlation technique was developed to detect this atypical fiber realignment using QPLI and mechanical data collected from human facet capsular ligaments (n = 16) loaded until visible rupture. Initial detection of anomalous realignment coincided with a measurable decrease in the tissue stiffness in every specimen and occurred at significantly lower strains than those at visible rupture (p<0.004), suggesting this technique may be sensitive to a loss of microstructural integrity. The spatial location of anomalous realignment was significantly associated with regions where visible rupture developed (p<0.001). This analysis technique provides a foundation to identify regional differences in soft tissue injury tolerances and relevant mechanical thresholds.

Select this Article		Reduction of background in optoacoustic image sequences obtained under tissue deformation Michael Jaeger, Lea Siegenthaler, Michael Kitz, and Martin Frenz J. Biomed. Opt. 14, 054011 (Sep 16, 2009); http://dx.doi.org/10.1117/1.3227038 \| Cited 2 times Online Publication Date: Sep 16, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract For real-time optoacoustic imaging of the human body, a linear array transducer and reflection mode optical irradiation is preferably used. Experimental outcomes however revealed that such a setup results in significant image background, which prevents imaging structures at the ultimate depth limited only by the optical attenuation of the irradiating light and the signal noise level. Various sources of image background, such as bulk tissue absorption, reconstruction artifacts, and backscattered ultrasound, could be identified. To overcome these limitations, we developed a novel method that results in significantly reduced background and increased imaging depth. For this purpose, we acquire, in parallel, a series of optoacoustic and echo-ultrasound images while the tissue sample is gradually deformed by an externally applied force. Optoacoustic signals and background signals are differently affected by the deformation and can thus be distinguished by image processing. This method takes advantage of a combined optoacoustic/echo-ultrasound device and has a strong potential for improving real-time optoacoustic imaging of deep tissue structures.

Select this Article		Broadband reflectance spectroscopy for establishing a quantitative metric of vascular leak using the Miles assay John McMurdy, Jonathan Reichner, Zara Mathews, Mary Markey, Sunny Intwala, and Gregory Crawford J. Biomed. Opt. 14, 054012 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3233654 \| Cited 1 time Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Monitoring the physiological effects of biological mediators on vascular permeability is important for identifying potential targets for antivascular leak therapy. This therapy is relevant to treatments for pulmonary edema and other disorders. Current methods of quantifying vascular leak are in vitro and do not allow repeated measurement of the same animal. Using an in vivo diffuse reflectance optical method allows pharmacokinetic analysis of candidate antileak molecules. Here, vascular leak is assessed in mice and rats by using the Miles assay and introducing irritation both topically using mustard oil and intradermally using vascular endothelial growth factor (VEGF). The severity of the leak is assessed using broadband diffuse reflectance spectroscopy with a fiber reflectance probe. Postprocessing techniques are applied to extract an artificial quantitative metric of leak from reflectance spectra at vascular leak sites on the skin of the animal. This leak metric is calculated with respect to elapsed time from irritation in both mustard oil and VEGF treatments on mice and VEGF treatments on rats, showing a repeatable increase in leak metric with leak severity. Furthermore, effects of pressure on the leak metric are observed to have minimal effect on the reflectance spectra, while spatial positioning showed spatially nonuniform leak sites.

Select this Article		Comparison of diffusion approximation and higher order diffusion equations for optical tomography of osteoarthritis Zhen Yuan, Qizhi Zhang, Eric Sobel, and Huabei Jiang J. Biomed. Opt. 14, 054013 (Oct 07, 2009); http://dx.doi.org/10.1117/1.3233655 \| Cited 3 times Online Publication Date: Oct 07, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract In this study, a simplified spherical harmonics approximated higher order diffusion model is employed for 3-D diffuse optical tomography of osteoarthritis in the finger joints. We find that the use of a higher-order diffusion model in a stand-alone framework provides significant improvement in reconstruction accuracy over the diffusion approximation model. However, we also find that this is not the case in the image-guided setting when spatial prior knowledge from x-rays is incorporated. The results show that the reconstruction error between these two models is about 15 and 4%, respectively, for stand-alone and image-guided frameworks.

Select this Article		Coregistered three-dimensional ultrasound and photoacoustic imaging system for ovarian tissue characterization Andres Aguirre, Puyun Guo, John Gamelin, Shikui Yan, Mary M. Sanders, Molly Brewer, and Quing Zhu J. Biomed. Opt. 14, 054014 (Sep 22, 2009); http://dx.doi.org/10.1117/1.3233916 \| Cited 10 times Online Publication Date: Sep 22, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Ovarian cancer has the highest mortality of all gynecologic cancers, with a five-year survival rate of only 30% or less. Current imaging techniques are limited in sensitivity and specificity in detecting early stage ovarian cancer prior to its widespread metastasis. New imaging techniques that can provide functional and molecular contrasts are needed to reduce the high mortality of this disease. One such promising technique is photoacoustic imaging. We develop a 1280-element coregistered 3-D ultrasound and photoacoustic imaging system based on a 1.75-D acoustic array. Volumetric images over a scan range of 80 deg in azimuth and 20 deg in elevation can be achieved in minutes. The system has been used to image normal porcine ovarian tissue. This is an important step toward better understanding of ovarian cancer optical properties obtained with photoacoustic techniques. To the best of our knowledge, such data are not available in the literature. We present characterization measurements of the system and compare coregistered ultrasound and photoacoustic images of ovarian tissue to histological images. The results show excellent coregistration of ultrasound and photoacoustic images. Strong optical absorption from vasculature, especially highly vascularized corpora lutea and low absorption from follicles, is demonstrated.

Select this Article		Control of optical contrast using gold nanoshells for optical coherence tomography imaging of mouse xenograft tumor model in vivo James Chen Yong Kah, Malini Olivo, Tzu Hao Chow, Kin San Song, Karen Zhen Yu Koh, Subodh Mhaisalkar, and Colin James Richard Sheppard J. Biomed. Opt. 14, 054015 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3233946 \| Cited 8 times Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The control of image contrast is essential toward optimizing a contrast enhancement procedure in optical coherence tomography (OCT). In this study, the in vivo control of optical contrast in a mouse tumor model with gold nanoshells as a contrast agent is examined. Gold nanoshells are administered into mice, with the injected dosage and particle surface parameters varied and its concentration in the tumor under each condition is determined using a noninvasive theoretical OCT modeling technique. The results show that too high a concentration of gold nanoshells in the tumor only enhances the OCT signal near the tissue surface, while significantly attenuating the signal deeper into the tissue. With an appropriate dosage, IV delivery of gold nanoshells allows a moderate concentration of 6.2×10⁹ particles/ml in tumor to achieve a good OCT signal enhancement with minimal signal attenuation with depth. An increase in the IV dosage of gold nanoshells reveals a corresponding nonlinear increase in their tumor concentration, as well as a nonlinear reduction in the fractional concentration of injected gold nanoshells. Furthermore, this fractional concentration is improved with the use of antiepodermal growth factor receptor (EGFR) surface functionalization, which also reduces the time required for tumor delivery from 6 to 2 h.

Select this Article		Investigation of the prefrontal cortex in response to duration-variable anagram tasks using functional near-infrared spectroscopy Fenghua Tian, Britton Chance, and Hanli Liu J. Biomed. Opt. 14, 054016 (Oct 07, 2009); http://dx.doi.org/10.1117/1.3241984 \| Cited 5 times Online Publication Date: Oct 07, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We hypothesize that nonlinearity between short-term anagram tasks and corresponding hemodynamic responses can be observed by functional near-infrared spectroscopy (fNIRS) in the prefrontal cortex (PFC). The PFC of six human subjects in response to anagram tasks is investigated using multichannel fNIRS. Concentration changes of oxyhemoglobin and deoxyhemoglobin in the PFC are measured with variable anagram durations and at two difficulty levels (four- and six-letter anagrams). The durations to perform the selected anagram tasks range from several seconds to more than one minute. The dorsolateral PFC areas exhibit consistent and strong hemodynamic deactivation during and shortly after task execution. The superposition principle of a linear system is employed to investigate nonlinear hemodynamic features among three task duration subgroups: D1 = 2.0 sec, D2 = 4.0 sec, and D3 = 8.0 sec. Such analysis shows clear nonlinearity in hemodynamic responses on the PFC with task durations shorter than 4 sec. Our observation of significant deactivation in early hemodynamic responses in the PFC is consistent with multiple fNIRS studies and several reports given in the field of functional magnetic resonance imaging. A better understanding of nonlinearity in fNIRS signals will have potential for us to investigate brain adaptation and to extrapolate neuronal activities from hemodynamic signals.

Select this Article OPEN ACCESS		Profile of new green fluorescent protein transgenic Jinhua pigs as an imaging source Tatsuo Kawarasaki, Kazuhiko Uchiyama, Atsushi Hirao, Sadahiro Azuma, Masayoshi Otake, Masatoshi Shibata, Seiko Tsuchiya, Shin Enosawa, Koichi Takeuchi, Kenjiro Konno, Yoji Hakamata, Hiroyuki Yoshino, Takuya Wakai, Shigeo Ookawara, Hozumi Tanaka, et al. J. Biomed. Opt. 14, 054017 (Oct 12, 2009); http://dx.doi.org/10.1117/1.3241985 \| Cited 6 times Online Publication Date: Oct 12, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Animal imaging sources have become an indispensable material for biological sciences. Specifically, gene-encoded biological probes serve as stable and high-performance tools to visualize cellular fate in living animals. We use a somatic cell cloning technique to create new green fluorescent protein (GFP)-expressing Jinhua pigs with a miniature body size, and characterized the expression profile in various tissues/organs and ex vivo culture conditions. The born GFP-transgenic pig demonstrate an organ/tissue-dependent expression pattern. Strong GFP expression is observed in the skeletal muscle, pancreas, heart, and kidney. Regarding cellular levels, bone-marrow-derived mesenchymal stromal cells, hepatocytes, and islet cells of the pancreas also show sufficient expression with the unique pattern. Moreover, the cloned pigs demonstrate normal growth and fertility, and the introduced GFP gene is stably transmitted to pigs in subsequent generations. The new GFP-expressing Jinhua pigs may be used as new cellular/tissue light resources for biological imaging in preclinical research fields such as tissue engineering, experimental regenerative medicine, and transplantation.

Select this Article OPEN ACCESS		Modeling optical behavior of birefringent biological tissues for evaluation of quantitative polarized light microscopy Mark C. van Turnhout, Sander Kranenbarg, and Johan L. van Leeuwen J. Biomed. Opt. 14, 054018 (Oct 12, 2009); http://dx.doi.org/10.1117/1.3241986 \| Cited 5 times Online Publication Date: Oct 12, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Quantitative polarized light microscopy (qPLM) is a popular tool for the investigation of birefringent architectures in biological tissues. Collagen, the most abundant protein in mammals, is such a birefringent material. Interpretation of results of qPLM in terms of collagen network architecture and anisotropy is challenging, because different collagen networks may yield equal qPLM results. We created a model and used the linear optical behavior of collagen to construct a Jones or Mueller matrix for a histological cartilage section in an optical qPLM train. Histological sections of tendon were used to validate the basic assumption of the model. Results show that information on collagen densities is needed for the interpretation of qPLM results in terms of collagen anisotropy. A parameter that is independent of the optical system and that measures collagen fiber anisotropy is introduced, and its physical interpretation is discussed. With our results, we can quantify which part of different qPLM results is due to differences in collagen densities and which part is due to changes in the collagen network. Because collagen fiber orientation and anisotropy are important for tissue function, these results can improve the biological and medical relevance of qPLM results.

Select this Article		Novel fluorescence molecular imaging of chemotherapy-induced intestinal apoptosis Galit Levin, Anat Shirvan, Hagit Grimberg, Ayelet Reshef, Merav Yogev-Falach, Avi Cohen, and Ilan Ziv J. Biomed. Opt. 14, 054019 (Oct 27, 2009); http://dx.doi.org/10.1117/1.3253303 Online Publication Date: Oct 27, 2009 Full Text: Read Online (HTML) \| Download PDF See Also: Erratum
	+ -	Show Abstract Chemotherapy-induced enteropathy (CIE) is one of the most serious complications of anticancer therapy, and tools for its early detection and monitoring are highly needed. We report on a novel fluorescence method for detection of CIE, based on molecular imaging of the related apoptotic process. The method comprises systemic intravenous administration of the ApoSense fluorescent biomarker (N,N′-didansyl-L-cystine DDC) in vivo and subsequent fluorescence imaging of the intestinal mucosa. In the reported proof-of-concept studies, mice were treated with either taxol+cyclophosphamide or doxil. DDC was administered in vivo at various time points after drug administration, and tracer uptake by ileum tissue was subsequently evaluated by ex vivo fluorescent microscopy. Chemotherapy caused marked and selective uptake of DDC in ileal epithelial cells, in correlation with other hallmarks of apoptosis (i.e., DNA fragmentation and Annexin-V binding). Induction of DDC uptake occurred early after chemotherapy, and its temporal profile was parallel to that of the apoptotic process, as assessed histologically. DDC may therefore serve as a useful tool for detection of CIE. Future potential integration of this method with fluorescent endoscopic techniques, or development of radio-labeled derivatives of DDC for emission tomography, may advance early diagnosis and monitoring of this severe adverse effect of chemotherapy.

Select this Article		Simultaneous three-dimensional optical coherence tomography and intravital microscopy for imaging subpleural pulmonary alveoli in isolated rabbit lungs Sven Meissner, Lilla Knels, Alexander Krueger, Thea Koch, and Edmund Koch J. Biomed. Opt. 14, 054020 (Oct 12, 2009); http://dx.doi.org/10.1117/1.3247149 \| Cited 11 times Online Publication Date: Oct 12, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract There is a growing interest in analyzing lung mechanics at the level of the alveoli in order to understand stress-related pathogenesis and possibly avoid ventilator associated lung injury. Emerging quantitative models to simulate fluid mechanics and the associated stresses and strains on delicate alveolar walls require realistic quantitative input on alveolar geometry and its dynamics during ventilation. Here, three-dimensional optical coherence tomography (OCT) and conventional intravital microscopy are joined in one setup to investigate the geometric changes of subpleural alveoli during stepwise pressure increase and release in an isolated and perfused rabbit lung model. We describe good qualitative agreement and quantitative correlation between the OCT data and video micrographs. Our main finding is the inflation and deflation of individual alveoli with noticeable hysteresis. Importantly, this three-dimensional geometry data can be extracted and converted into input data for numerical simulations.

Select this Article		Intra-operative brain tumor detection using elastic light single-scattering spectroscopy: a feasibility study Murat Canpolat, Mahmut Akyüz, Güzide Ayşe Gökhan, Elif İnanç Gürer, and Recai Tuncer J. Biomed. Opt. 14, 054021 (Oct 14, 2009); http://dx.doi.org/10.1117/1.3247151 \| Cited 4 times Online Publication Date: Oct 14, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We have investigated the potential application of elastic light single-scattering spectroscopy (ELSSS) as an adjunctive tool for intraoperative rapid detection of brain tumors and demarcation of the tumor from the surrounding normal tissue. Measurements were performed on 29 excised tumor specimens from 29 patients. There were 21 instances of low-grade tumors and eight instances of high-grade tumors. Normal gray matter and white matter brain tissue specimens of four epilepsy patients were used as a control group. One low-grade and one high-grade tumor were misclassified as normal brain tissue. Of the low- and high-grade tumors, 20 out of 21 and 7 out of 8 were correctly classified by the ELSSS system, respectively. One normal white matter tissue margin was detected in a high-grade tumor, and three normal tissue margins were detected in three low-grade tumors using spectroscopic data analysis and confirmed by histopathology. The spectral slopes were shown to be positive for normal white matter brain tissue and negative for normal gray matter and tumor tissues. Our results indicate that signs of spectral slopes may enable the discrimination of brain tumors from surrounding normal white matter brain tissue with a sensitivity of 93% and specificity of 100%.

Select this Article OPEN ACCESS		Hemodynamic and electrophysiological signals of conflict processing in the Chinese-character Stroop task: a simultaneous near-infrared spectroscopy and event-related potential study Jiahuan Zhai, Ting Li, Zhongxing Zhang, and Hui Gong J. Biomed. Opt. 14, 054022 (Oct 29, 2009); http://dx.doi.org/10.1117/1.3247152 \| Cited 1 time Online Publication Date: Oct 29, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract A dual-modality method combining continuous-wave near-infrared spectroscopy (NIRS) and event-related potentials (ERPs) was developed for the Chinese-character color-word Stroop task, which included congruent, incongruent, and neutral stimuli. Sixteen native Chinese speakers participated in this study. Hemodynamic and electrophysiological signals in the prefrontal cortex (PFC) were monitored simultaneously by NIRS and ERP. The hemodynamic signals were represented by relative changes in oxy-, deoxy-, and total hemoglobin concentration, whereas the electrophysiological signals were characterized by the parameters P450, N500, and P600. Both types of signals measured at four regions of the PFC were analyzed and compared spatially and temporally among the three different stimuli. We found that P600 signals correlated significantly with the hemodynamic parameters, suggesting that the PFC executes conflict-solving function. Additionally, we observed that the change in deoxy-Hb concentration showed higher sensitivity in response to the Stroop task than other hemodynamic signals. Correlation between NIRS and ERP signals revealed that the vascular response reflects the cumulative effect of neural activities. Taken together, our findings demonstrate that this new dual-modality method is a useful approach to obtaining more information during cognitive and physiological studies.

Select this Article OPEN ACCESS		Diagnosing breast cancer using Raman spectroscopy: prospective analysis Abigail S. Haka, Zoya Volynskaya, Joseph A. Gardecki, Jon Nazemi, Robert Shenk, Nancy Wang, Ramachandra R. Dasari, Maryann Fitzmaurice, and Michael S. Feld J. Biomed. Opt. 14, 054023 (Oct 14, 2009); http://dx.doi.org/10.1117/1.3247154 \| Cited 14 times Online Publication Date: Oct 14, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We present the first prospective test of Raman spectroscopy in diagnosing normal, benign, and malignant human breast tissues. Prospective testing of spectral diagnostic algorithms allows clinicians to accurately assess the diagnostic information contained in, and any bias of, the spectroscopic measurement. In previous work, we developed an accurate, internally validated algorithm for breast cancer diagnosis based on analysis of Raman spectra acquired from fresh-frozen in vitro tissue samples. We currently evaluate the performance of this algorithm prospectively on a large ex vivo clinical data set that closely mimics the in vivo environment. Spectroscopic data were collected from freshly excised surgical specimens, and 129 tissue sites from 21 patients were examined. Prospective application of the algorithm to the clinical data set resulted in a sensitivity of 83%, a specificity of 93%, a positive predictive value of 36%, and a negative predictive value of 99% for distinguishing cancerous from normal and benign tissues. The performance of the algorithm in different patient populations is discussed. Sources of bias in the in vitro calibration and ex vivo prospective data sets, including disease prevalence and disease spectrum, are examined and analytical methods for comparison provided.

Select this Article		Thermoacoustic and photoacoustic sensing of temperature Manojit Pramanik and Lihong V. Wang J. Biomed. Opt. 14, 054024 (Oct 12, 2009); http://dx.doi.org/10.1117/1.3247155 \| Cited 16 times Online Publication Date: Oct 12, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We present a novel temperature-sensing technique using thermoacoustic and photoacoustic measurements. This noninvasive method has been demonstrated using a tissue phantom to have high temporal resolution and temperature sensitivity. Because both photoacoustic and thermoacoustic signal amplitudes depend on the temperature of the source object, the signal amplitudes can be used to monitor the temperature. A temperature sensitivity of 0.15°C was obtained at a temporal resolution as short as 2 s, taking the average of 20 signals. The deep-tissue imaging capability of this technique can potentially lead us to in vivo temperature monitoring in thermal or cryogenic applications.

Select this Article		Risk assessment of the application of a plasma jet in dermatology Juergen Lademann, Heike Richter, Alena Alborova, Daniel Humme, Alexa Patzelt, Axel Kramer, Klaus-Dieter Weltmann, Bernd Hartmann, Christian Ottomann, Joachim W. Fluhr, Peter Hinz, Georg Hübner, and Olaf Lademann J. Biomed. Opt. 14, 054025 (Oct 14, 2009); http://dx.doi.org/10.1117/1.3247156 \| Cited 11 times Online Publication Date: Oct 14, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Regardless of the fact that several highly efficient antiseptics are commercially available, the antiseptic treatment of chronic wounds remains a problem. In the past, electrical plasma discharges have been frequently used in biometrical science for disinfection and sterilization of material surfaces. Plasma systems usually have a temperature of several hundred degrees. Recently, it was reported that “cold” plasma can be applied onto living tissue. In in vitro studies on cell culture, it could be demonstrated that this new plasma possesses excellent antiseptic properties. We perform a risk assessment concerning the in vivo application of a “cold” plasma jet on patients and volunteers. Two potential risk factors, UV radiation and temperature, are evaluated. We show that the UV radiation of the plasma in the used system is an order of magnitude lower than the minimal erythema dose, necessary to produce sunburn on the skin in vivo. Additionally, thermal damage of the tissue by the plasma can be excluded. The results of the risk assessment stimulate the in vivo application of the investigated plasma jet in the treatment of chronic wounds.

Select this Article		Single-wavelength reflected confocal and multiphoton microscopy for tissue imaging Wei-Liang Chen, Chen-Kuan Chou, Ming-Gu Lin, Yang-Fang Chen, Shiou-Hwa Jee, Hsin-Yuan Tan, Tsung-Hua Tsai, Ki-Hean Kim, Daekeun Kim, Peter T. C. So, Sung-Jan Lin, and Chen-Yuan Dong J. Biomed. Opt. 14, 054026 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3247157 \| Cited 1 time Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Both reflected confocal and multiphoton microscopy can have clinical diagnostic applications. The successful combination of both modalities in tissue imaging enables unique image contrast to be achieved, especially if a single laser excitation wavelength is used. We apply this approach for skin and corneal imaging using the 780-nm output of a femtosecond, titanium-sapphire laser. We find that the near-IR, reflected confocal (RC) signal is useful in characterizing refractive index varying boundaries in bovine cornea and porcine skin, while the multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) intensities can be used to image cytoplasm and connective tissues (collagen), respectively. In addition, quantitative analysis shows that we are able to detect MAF from greater imaging depths than with the near-IR RC signal. Furthermore, by performing RC imaging at 488, 543, and 633 nm, we find that a longer wavelength leads to better image contrast for deeper imaging of the bovine cornea and porcine skin tissue. Finally, by varying power of the 780-nm source, we find that comparable RC image quality was achieved in the 2.7 to 10.7-mW range.

Select this Article		Noninvasive assessment of testicular torsion in rabbits using frequency-domain near-infrared spectroscopy: prospects for pediatric urology Bertan Hallacoglu, Richard S. Matulewicz, Harriet J. Paltiel, Horacio Padua, Patricio Gargollo, Glenn Cannon, Ahmad Alomari, Angelo Sassaroli, and Sergio Fantini J. Biomed. Opt. 14, 054027 (Oct 28, 2009); http://dx.doi.org/10.1117/1.3253318 \| Cited 3 times Online Publication Date: Oct 28, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We present a quantitative near-IR spectroscopy study of the absolute values of oxygen saturation of hemoglobin before and after surgically induced testicular torsion in adult rabbits. Unilateral testicular torsions (0, 540, or 720 deg) on experimental testes and contralateral sham surgery on control testes are performed in four adult rabbits. A specially designed optical probe for measurements at multiple source-detector distances and a commercial frequency-domain tissue spectrometer are used to measure absolute values of testicular hemoglobin saturation. Our results show: (1) a consistent baseline absolute tissue hemoglobin saturation value of 78±5%, (2) a comparable tissue hemoglobin saturation of 77±6% after sham surgery, and (3) a significantly lower tissue hemoglobin saturation of 36±2% after 540- and 720-deg testicular torsion surgery. Our findings demonstrate the feasibility of performing frequency-domain, multidistance near-IR spectroscopy for absolute testicular oximetry in the assessment of testicular torsion. We conclude that near-IR spectroscopy has potential to serve as a clinical diagnostic and monitoring tool for the assessment of absolute testicular hemoglobin desaturation caused by torsion, with the possibility of serving as a complement to conventional color and spectral Doppler ultrasonography.

Select this Article		Study of cadmium-induced cytotoxicity using two-photon excitation endogenous fluorescence microscopy Dong Li, Mildred S. Yang, Tao Lin, Wei Zheng, and Jianan Y. Qu J. Biomed. Opt. 14, 054028 (Oct 23, 2009); http://dx.doi.org/10.1117/1.3250293 \| Cited 1 time Online Publication Date: Oct 23, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We demonstrate that using time-resolved two-photon excitation endogenous fluorescence microscopy, the cadmium (Cd)-induced cellular toxic level can be assessed by the free-to protein-bound reduced nicotinamide adenine dinucleotide (free/bound NADH) ratio in a living cell. NADH fluorescence excited at 730 nm is captured at different times following exposure to cadmium at a variety of concentrations. The temporal characteristics of NADH fluorescence from mitochondrial and nuclear compartments are analyzed, respectively. The results show that cadmium induces a significant increase of the free/bound NADH ratio in mitochondria and nucleus, caused by the inhibition effect on the electron transport chain (ETC) and the stimulating effect on the glycolysis pathway, respectively. It is found that induction of metallothionein (MT) in cells occurs after 4 h of exposure to a sublethal concentration of Cd and reaches a peak at 6 h. More importantly, the increase in MT level can effectively suppress the elevation of the free/bound NADH ratio caused by a subsequent exposure to a higher concentration of Cd, indicating that MT plays a key role in protecting cells from Cd-induced toxicity. Our findings show that the free/bound NADH ratio can potentially be used as a sensitive indicator of toxic and carcinogenic actions induced by Cd.

Select this Article		Optical properties of dental restorative materials in the wavelength range 400 to 700 nm for the simulation of color perception Moritz Friebel, Kirsten Povel, Hans-Joachim Cappius, Jürgen Helfmann, and Martina Meinke J. Biomed. Opt. 14, 054029 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3250292 \| Cited 1 time Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Aesthetic restorations require dental restorative materials to have optical properties very similar to those of the teeth. A method is developed to this end to determine the optical parameters absorption coefficient μ_a, scattering coefficient μ_s, anisotropy factor g, and effective scattering coefficient μs′ of dental restorative materials. The method includes sample preparation and measurements of transmittance and reflectance in an integrating sphere spectrometer followed by inverse Monte Carlo simulations. Using this method the intrinsic optical parameters are determined for shade B2 of the light-activated composites TPH^® Spectrum^®, Esthet-X^®, and the Ormocer^® Definite^® in the wavelength range 400 to 700 nm. By using the determined parameters μ_a, μ_s, and g together with an appropriate phase function, the reflectance of samples with 1-mm layer thickness and shade B2 could be predicted with a very high degree of accuracy using a forward Monte Carlo simulation. The color perception was calculated from the simulated reflectance according to the CIELAB system. We initiate the compilation of a data pool of optical parameters that in the future will enable calculation models to be used as a basis for optimization of the optical approximation of the natural tooth, and the composition of new materials and their production process.

Select this Article OPEN ACCESS		Diffuse optical spectroscopy of breast tissue extended to 1100 nm Paola Taroni, Andrea Bassi, Daniela Comelli, Andrea Farina, Rinaldo Cubeddu, and Antonio Pifferi J. Biomed. Opt. 14, 054030 (Oct 28, 2009); http://dx.doi.org/10.1117/1.3251051 \| Cited 10 times Online Publication Date: Oct 28, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The feasibility of in vivo measurements in the range of 1000 to 1100 nm and the potential benefits of operation in that wavelength range for diagnostic applications are investigated. To this purpose, an existing system for time-resolved diffuse spectroscopy is modified to enable in vivo studies to be carried out continuously from 600 to 1100 nm. The optical characterization of collagen powder is extended to 1100 nm and an accurate measurement of the absorption properties of lipid is carried out over the entire spectral range. Finally, the first in vivo absorption and scattering spectra of breast tissue are measured from 10 healthy volunteers between 600 and 1100 nm and tissue composition is evaluated in terms of blood parameters and water, lipid, and collagen content using a spectrally constrained global fitting procedure.

Select this Article OPEN ACCESS		Development of Raman microspectroscopy for automated detection and imaging of basal cell carcinoma Marta Larraona-Puy, Adrian Ghita, Alina Zoladek, William Perkins, Sandeep Varma, Iain H. Leach, Alexey A. Koloydenko, Hywel Williams, and Ioan Notingher J. Biomed. Opt. 14, 054031 (Oct 27, 2009); http://dx.doi.org/10.1117/1.3251053 \| Cited 8 times Online Publication Date: Oct 27, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We investigate the potential of Raman microspectroscopy (RMS) for automated evaluation of excised skin tissue during Mohs micrographic surgery (MMS). The main aim is to develop an automated method for imaging and diagnosis of basal cell carcinoma (BCC) regions. Selected Raman bands responsible for the largest spectral differences between BCC and normal skin regions and linear discriminant analysis (LDA) are used to build a multivariate supervised classification model. The model is based on 329 Raman spectra measured on skin tissue obtained from 20 patients. BCC is discriminated from healthy tissue with 90±9% sensitivity and 85±9% specificity in a 70% to 30% split cross-validation algorithm. This multivariate model is then applied on tissue sections from new patients to image tumor regions. The RMS images show excellent correlation with the gold standard of histopathology sections, BCC being detected in all positive sections. We demonstrate the potential of RMS as an automated objective method for tumor evaluation during MMS. The replacement of current histopathology during MMS by a “generalization” of the proposed technique may improve the feasibility and efficacy of MMS, leading to a wider use according to clinical need.

Select this Article OPEN ACCESS		Comparison of principal and independent component analysis in removing extracerebral interference from near-infrared spectroscopy signals Jaakko Virtanen, Tommi Noponen, and Pekka Meriläinen J. Biomed. Opt. 14, 054032 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253323 \| Cited 13 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Near-infrared spectroscopy (NIRS) is a method for noninvasive estimation of cerebral hemodynamic changes. Principal component analysis (PCA) and independent component analysis (ICA) can be used for decomposing a set of signals to underlying components. Our objective is to determine whether PCA or ICA is more efficient in identifying and removing scalp blood flow interference from multichannel NIRS signals. Concentration changes of oxygenated (HbO₂) and deoxygenated (HbR) hemoglobin are measured on the forehead with multichannel NIRS during hyper- and hypocapnia. PCA and ICA are used separately to identify and remove signal contribution from extracerebral tissue, and the resulting estimates of cerebral responses are compared to the expected cerebral responses. Both methods were able to reduce extracerebral contribution to the signals, but PCA typically performs equal to or better than ICA. The improvement in 3-cm signal quality achieved with both methods is comparable to increasing the source-detector separation from 3 to 5 cm. Especially PCA appears to be well suited for use in NIRS applications where the cerebral activation is diffuse, such as monitoring of global cerebral oxygenation and hemodynamics. Performance differences between PCA and ICA could be attributed primarily to different criteria for identifying the surface effect.

Select this Article OPEN ACCESS		Fiber evanescent wave spectroscopy using the mid-infrared provides useful fingerprints for metabolic profiling in humans Marie-Laure Anne, Caroline Le Lan, Valérie Monbet, Catherine Boussard-Plédel, Martine Ropert, Olivier Sire, Michel Pouchard, Christine Jard, Jacques Lucas, Jean Luc Adam, Pierre Brissot, Bruno Bureau, and Olivier Loréal J. Biomed. Opt. 14, 054033 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253319 \| Cited 4 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Fiber evanescent wave spectroscopy (FEWS) explores the mid-infrared domain, providing information on functional chemical groups represented in the sample. Our goal is to evaluate whether spectral fingerprints obtained by FEWS might orientate clinical diagnosis. Serum samples from normal volunteers and from four groups of patients with metabolic abnormalities are analyzed by FEWS. These groups consist of iron overloaded genetic hemochromatosis (GH), iron depleted GH, cirrhosis, and dysmetabolic hepatosiderosis (DYSH). A partial least squares (PLS) logistic method is used in a training group to create a classification algorithm, thereafter applied to a test group. Patients with cirrhosis or DYSH, two groups exhibiting important metabolic disturbances, are clearly discriminated from control groups with AUROC values of 0.94±0.05 and 0.90±0.06, and sensibility/specificity of 86/84% and 87/87%, respectively. When pooling all groups, the PLS method contributes to discriminate controls, cirrhotic, and dysmetabolic patients. Our data demonstrate that metabolic profiling using infrared FEWS is a possible way to investigate metabolic alterations in patients.

Select this Article		Influence of laser parameters on nanoparticle-induced membrane permeabilization Cuiping Yao, Xiaochao Qu, Zhenxi Zhang, Gereon Hüttmann, and Ramtin Rahmanzadeh J. Biomed. Opt. 14, 054034 (Oct 27, 2009); http://dx.doi.org/10.1117/1.3253320 \| Cited 10 times Online Publication Date: Oct 27, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Light-absorbing nanoparticles that are heated by short laser pulses can transiently increase membrane permeability. We evaluate the membrane permeability by flow cytometry assaying of propidium iodide and fluorescein isothiocyanate dextran (FITC-D) using different laser sources. The dependence of the transfection efficiency on laser parameters such as pulse duration, irradiant exposure, and irradiation mode is investigated. For nano- and also picosecond irradiation, we show a parameter range where a reliable membrane permeabilization is achieved for 10-kDa FITC-D. Fluorescent labeled antibodies are able to penetrate living cells that are permeabilized using these parameters. More than 50% of the cells are stained positive for a 150-kDa IgG antibody. These results suggest that the laser-induced permeabilization approach constitutes a promising tool for targeted delivery of larger exogenous molecules into living cells.

Select this Article OPEN ACCESS		Optical anisotropic diffusion: new model systems and theoretical modeling Patrick M. Johnson and Ad Lagendijk J. Biomed. Opt. 14, 054036 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253332 \| Cited 2 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We have developed several new experimental model systems that demonstrate anisotropic diffusion of light. These systems, consisting of aligned fibers, stretched plastic foam, and stretched plastic frit, have relatively simple microstructures and are easily sliced, making them ideal for testing theoretical models of diffusion. We demonstrate that the solution to the diffusion equation for arbitrary orientation of the diffusion tensor is consistent with experimental measurements. We also show that simple models of microstructure, based on cylindrical and planar scatterers, are consistent with the experimental results. These models provide simple analytical expressions for predicting the degree of alignment of the scatterers from diffuse transmission measurements. The combination of experimental results and theoretical support demonstrates both the power and the limitations of the diffusion model for providing information about microstructure via simple experiments and modeling.

Select this Article OPEN ACCESS		Method for optical coherence tomography image classification using local features and earth mover’s distance Yankui Sun and Ming Lei J. Biomed. Opt. 14, 054037 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3251059 Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Optical coherence tomography (OCT) is a recent imaging method that allows high-resolution, cross-sectional imaging through tissues and materials. Over the past 18 years, OCT has been successfully used in disease diagnosis, biomedical research, material evaluation, and many other domains. As OCT is a recent imaging method, until now surgeons have limited experience using it. In addition, the number of images obtained from the imaging device is too large, so we need an automated method to analyze them. We propose a novel method for automated classification of OCT images based on local features and earth mover’s distance (EMD). We evaluated our algorithm using an OCT image set which contains two kinds of skin images, normal skin and nevus flammeus. Experimental results demonstrate the effectiveness of our method, which achieved classification accuracy of 0.97 for an EMD+KNN scheme and 0.99 for an EMD+SVM (support vector machine) scheme, much higher than the previous method. Our approach is especially suitable for nonhomogeneous images and could be applied to a wide range of OCT images.

Select this Article OPEN ACCESS		New method of estimating wavelength-dependent optical path length ratios for oxy- and deoxyhemoglobin measurement using near-infrared spectroscopy Shinji Umeyama and Toru Yamada J. Biomed. Opt. 14, 054038 (Oct 27, 2009); http://dx.doi.org/10.1117/1.3253350 \| Cited 2 times Online Publication Date: Oct 27, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract In near-infrared spectroscopy (NIRS), concentration changes in oxy- and deoxyhemoglobin are calculated using an attenuation change of the measurement light and by solving a linear equation based on the modified Lambert-Beer law. While solving this equation, we need to know the wavelength-dependent mean optical path lengths of the measurement lights. However, it is very difficult to know these values by a continuous-wave-type (CW-type) system. We propose a new method of estimating wavelength-dependent optical path length ratios of the measurement lights based on the data obtained by a triple wavelength CW-type NIRS instrument. The proposed method does not give a path length itself, but it gives a path length ratio. Thus, it is possible to obtain the accurate hemoglobin concentration changes without cross talk, although the method cannot contribute to the quantification of the absolute magnitude of hemoglobin changes. The method is based on the principle that two possible estimations of hemoglobin concentration changes calculated using a triple-wavelength measurement system should be identical. The method was applied to the experimental data of human subjects’ foreheads. The estimated path length ratios were very similar to literature values obtained by using picosecond laser pulses and a streak camera detector [ M. Essenpreis et al., Appl. Opt. 32(4), 418–425 (1993) ].

Select this Article OPEN ACCESS		Comparison of spectroscopically measured tissue alcohol concentration to blood and breath alcohol measurements Trent D. Ridder, Benjamin J. Ver Steeg, and Bentley D. Laaksonen J. Biomed. Opt. 14, 054039 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253353 \| Cited 1 time Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Alcohol testing is an expanding area of interest due to the impacts of alcohol abuse that extend well beyond drunk driving. However, existing approaches such as blood and urine assays are hampered in some testing environments by biohazard risks. A noninvasive, in vivo spectroscopic technique offers a promising alternative, as no body fluids are required. The purpose of this work is to report the results of a 36-subject clinical study designed to characterize tissue alcohol measured using near-infrared spectroscopy relative to venous blood, capillary blood, and breath alcohol. Comparison of blood and breath alcohol concentrations demonstrated significant differences in alcohol concentration [root mean square of 9.0 to 13.5 mg/dL] that were attributable to both assay accuracy and precision as well as alcohol pharmacokinetics. A first-order kinetic model was used to estimate the contribution of alcohol pharmacokinetics to the differences in concentration observed between the blood, breath, and tissue assays. All pair-wise combinations of alcohol assays were investigated, and the fraction of the alcohol concentration variance explained by pharmacokinetics ranged from 41.0% to 83.5%. Accounting for pharmacokinetic concentration differences, the accuracy and precision of the spectroscopic tissue assay were found to be comparable to those of the blood and breath assays.

Select this Article OPEN ACCESS		Repetition rate dependency of reactive oxygen species formation during femtosecond laser–based cell surgery Judith Baumgart, Kai Kuetemeyer, Willem Bintig, Anaclet Ngezahayo, Wolfgang Ertmer, Holger Lubatschowski, and Alexander Heisterkamp J. Biomed. Opt. 14, 054040 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253382 \| Cited 6 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF See Also: Erratum
	+ -	Show Abstract Femtosecond (fs) laser–based cell surgery is typically done in two different regimes, at kHz or MHz repetition rate. Formation of reactive oxygen species (ROS) is an often predicted effect due to illumination with short laser pulses in biological tissue. We present our study on ROS formation in single cells in response to irradiation with fs laser pulses depending on the repetition rate while focusing into the cell nucleus. We observed a significant increase of ROS concentration directly after manipulation followed by a decrease in both regimes at kHz and MHz repetition rate. In addition, effects of consecutive exposures at MHz and kHz repetition rate and vice versa on ROS production were studied. Irradiation with a MHz pulse train followed by a kHz pulse train resulted in a significantly higher increase of ROS concentration than in the reversed case and often caused cell death. In the presence of the antioxidant ascorbic acid, accumulation of ROS and cell death were strongly reduced. Therefore, addition of antioxidants during fs laser–based cell surgery experiments could be advantageous in terms of suppressing photochemical damage to the cell.

Select this Article OPEN ACCESS		Beyond the 1/T_p limit: two-photon-excited fluorescence using pulses as short as sub-10-fs Shuo Pang, Alvin T. Yeh, Chao Wang, and Kenith E. Meissner J. Biomed. Opt. 14, 054041 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253388 \| Cited 3 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Two-photon-excited fluorescence and second-harmonic generation are characterized as a function of laser pulse duration as short as sub-10-fs. A comparative study is performed where pulse duration is varied by introducing dispersion, as reported previously, and by tailoring pulse spectral width and minimizing its time-bandwidth product (transform-limited pulses). Experimental data and calculations show that characterizing a two-photon signal with the two schemes to vary pulse duration measures different phenomena. Two-photon signal characterization using dispersion-broadened pulses measures only the effect of chirp on the pulse two-photon-excitation spectrum and is independent of molecular response. Transform-limited pulses are used to measure the dependence of two-photon signal generation on pulse duration. Calculations show that deviation from the 1/T_p relationship would be expected as the transform-limited pulse spectral width approaches the molecular two-photon absorption linewidth and exhibits asymptotic behavior for pulse spectral widths 10 times greater than the absorption linewidth. Experimental measurements are consistent with the predicted behavior. The impact of using ultrashort laser pulses on the performance characteristics of nonlinear optical microscopy is discussed.

Select this Article OPEN ACCESS		Specific immobilization of influenza A virus on GaAs (001) surface Valerie Duplan, Yannick Miron, Eric Frost, Michel Grandbois, and Jan J. Dubowski J. Biomed. Opt. 14, 054042 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3251057 \| Cited 3 times Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract In the quest for the development of an all-optical biosensor for rapid detection and typing of viral pathogens, we investigate biosensing architectures that take advantage of strong photoluminescence emission from III-V quantum semiconductors (QS). One of the key elements in the development of such a biosensor is the ability to attach various analytes to GaAs—a material of choice for capping III-V QS of our interest. We report on the study of biofunctionalization of GaAs (001) with polyethylene-glycol (PEG) thiols and the successful immobilization of influenza A virus. A diluted solution of biotinylated PEG thiols in OH-terminated PEG thiols is used to form a network of sites for the attachment of neutravidin. Biotinylated polyclonal influenza A antibodies are applied to investigate the process of the immobilization of inactivated influenza A virus. The successful immobilization is demonstrated using atomic force microscopy and fluorescence microscopy measurements.

Select this Article OPEN ACCESS		Investigation of a probe design for facilitating the uses of the standard photon diffusion equation at short source-detector separations: Monte Carlo simulations Sheng-Hao Tseng, Carole Hayakawa, Jerome Spanier, and Anthony J. Durkin J. Biomed. Opt. 14, 054043 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253386 \| Cited 3 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract We design a special diffusing probe to investigate the optical properties of human skin in vivo. The special geometry of the probe enables a modified two-layer (MTL) diffusion model to precisely describe the photon transport even when the source-detector separation is shorter than 3 mean free paths. We provide a frequency domain comparison between the Monte Carlo model and the diffusion model in both the MTL geometry and conventional semiinfinite geometry. We show that using the Monte Carlo model as a benchmark method, the MTL diffusion theory performs better than the diffusion theory in the semiinfinite geometry. In addition, we carry out Monte Carlo simulations with the goal of investigating the dependence of the interrogation depth of this probe on several parameters including source-detector separation, sample optical properties, and properties of the diffusing high-scattering layer. From the simulations, we find that the optical properties of samples modulate the interrogation volume greatly, and the source-detector separation and the thickness of the diffusing layer are the two dominant probe parameters that impact the interrogation volume. Our simulation results provide design guidelines for a MTL geometry probe.

Select this Article OPEN ACCESS		Photometric measurements of red blood cell aggregation: light transmission versus light reflectance Oguz K. Baskurt, Mehmet Uyuklu, Max R. Hardeman, and Herbert J. Meiselman J. Biomed. Opt. 14, 054044 (Oct 23, 2009); http://dx.doi.org/10.1117/1.3251050 \| Cited 2 times Online Publication Date: Oct 23, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Red blood cell (RBC) aggregation is the reversible and regular clumping in the presence of certain macromolecules. This is a clinically important phenomenon, being significantly enhanced in the presence of acute phase reactants (e.g., fibrinogen). Both light reflection (LR) and light transmission (LT) from or through thin layers of RBC suspensions during the process of aggregation are accepted to reflect the time course of aggregation. It has been recognized that the time courses of LR and LT might be different from each other. We aim to compare the RBC aggregation measurements based on simultaneous recordings of LR and LT. The results indicate that LR during RBC aggregation is characterized by a faster time course compared to simultaneously recorded LT. This difference in time course of LR and LT is reflected in the calculated parameters reflecting the overall extent and kinetics of RBC aggregation. Additionally, the power of parameters calculated using LR and LT time courses in detecting a given difference in aggregation are significantly different from each other. These differences should be taken into account in selecting the appropriate calculated parameters for analyzing LR or LT time courses for the assessment of RBC aggregation.

Select this Article OPEN ACCESS		Near-infrared image-guided laser ablation of dental decay You-Chen Tao and Daniel Fried J. Biomed. Opt. 14, 054045 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253390 \| Cited 2 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Image-guided laser ablation systems are now feasible for dentistry with the recent development of nondestructive high-contrast imaging modalities such as near-IR (NIR) imaging and optical coherence tomography (OCT) that are capable of discriminating between sound and demineralized dental enamel at the early stages of development. Our objective is to demonstrate that images of demineralized tooth surfaces have sufficient contrast to be used to guide a CO₂ laser for the selective removal of natural and artificial caries lesions. NIR imaging and polarization-sensitive optical coherence tomography (PS-OCT) operating at 1310-nm are used to acquire images of natural lesions on extracted human teeth and highly patterned artificial lesions produced on bovine enamel. NIR and PS-OCT images are analyzed and converted to binary maps designating the areas on the samples to be removed by a CO₂ laser to selectively remove the lesions. Postablation NIR and PS-OCT images confirmed preferential removal of demineralized areas with minimal damage to sound enamel areas. These promising results suggest that NIR and PS-OCT imaging systems can be integrated with a CO₂ laser ablation system for the selective removal of dental caries.

Select this Article		Noncontact, nondestructive elasticity evaluation of sound and demineralized human dental enamel using a laser ultrasonic surface wave dispersion technique Hsiao-Chuan Wang, Simon Fleming, Yung-Chun Lee, Susan Law, Michael Swain, and Jing Xue J. Biomed. Opt. 14, 054046 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3253396 \| Cited 2 times Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Laser ultrasonic nondestructive evaluation (NDE) methods have been proposed to replace conventional in vivo dental clinical diagnosis tools that are either destructive or incapable of quantifying the elasticity of human dental enamel. In this work, a laser NDE system that can perform remote measurements on samples of small dimensions is presented. A focused laser line source is used to generate broadband surface acoustic wave impulses that are detected with a simplified optical fiber interferometer. The measured surface wave velocity dispersion spectrum is in turn used to characterize the elasticity of the specimen. The NDE system and the analysis technique are validated with measurements of different metal structures and then applied to evaluate human dental enamel. Artificial lesions are prepared on the samples to simulate different states of enamel elasticity. Measurement results for both sound and lesioned regions, as well as lesions of different severity, are clearly distinguishable from each other and fit well with physical expectations and theoretical value. This is the first time, to the best of our knowledge, that a laser-based surface wave velocity dispersion technique is successfully applied on human dental enamel, demonstrating the potential for noncontact, nondestructive in vivo detection of the development of carious lesions.

Select this Article		Depth-resolved measurement of ocular fundus pulsations by low-coherence tissue interferometry Nikolaus Dragostinoff, René M. Werkmeister, Martin Gröschl, and Leopold Schmetterer J. Biomed. Opt. 14, 054047 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3251049 \| Cited 2 times Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract A device that allows for the measurement of ocular fundus pulsations at preselected axial positions of a subject’s eye is presented. Unlike previously presented systems, which only allow for observation of the strongest reflecting retinal layer, our system enables the measurement of fundus pulsations at a preselected ocular layer. For this purpose the sample is illuminated by light of low temporal coherence. The layer is then selected by positioning one mirror of a Michelson interferometer according to the depth of the layer. The device contains a length measurement system based on partial coherence interferometry and a line scan charge-coupled device camera for recording and online inspection of the fringe system. In-vivo measurements in healthy humans are performed as proof of principle. The algorithms used for enhancing the recorded images are briefly introduced. The contrast of the observed interference pattern is evaluated for different positions of the measurement mirror and at various distances from the front surface of the cornea. The applications of such a system may be wide, including assessment of eye elongation during myopia development and blood-flow-related changes in intraocular volume.

Select this Article		Thermal imaging method for estimating oxygen saturation Michal Tepper, Rotem Neeman, Yonat Milstein, Moshe Ben David, and Israel Gannot J. Biomed. Opt. 14, 054048 (Oct 23, 2009); http://dx.doi.org/10.1117/1.3251036 \| Cited 1 time Online Publication Date: Oct 23, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The objective of this study is to develop a minimal invasive thermal imaging method to determine the oxygenation level of an internal tissue. In this method, the tissue is illuminated using an optical fiber by several wavelengths in the visible and near-IR range. Each wavelength is absorbed by the tissue and thus causes increase in its temperature. The temperature increase is observed by a coherent waveguide bundle in the mid-IR range. The thermal imaging of the tissue is done using a thermal camera through the coherent bundle. Analyzing the temperature rise allows estimating the tissue composition in general, and specifically the oxygenation level. Such a system enables imaging of the temperature within body cavities through a commercial endoscope. As an intermediate stage, the method is applied and tested on exposed skin tissue. A curve-fitting algorithm is used to find the most suitable saturation value affecting the temperature function. The algorithm is tested on a theoretical tissue model with various parameters, implemented for this study, and on agar phantom models. The calculated saturation values are in agreement with the real saturation values.

Select this Article		Evans blue dye-enhanced capillary-resolution photoacoustic microscopy in vivo Junjie Yao, Konstantin Maslov, Song Hu, and Lihong V. Wang J. Biomed. Opt. 14, 054049 (Oct 26, 2009); http://dx.doi.org/10.1117/1.3251044 \| Cited 7 times Online Publication Date: Oct 26, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Complete and continuous imaging of microvascular networks is crucial for a wide variety of biomedical applications. Photoacoustic tomography can provide high resolution microvascular imaging using hemoglobin within red blood cells (RBCs) as an endogenic contrast agent. However, intermittent RBC flow in capillaries results in discontinuous and fragmentary capillary images. To overcome this problem, we use Evans blue (EB) dye as a contrast agent for in vivo photoacoustic imaging. EB has strong optical absorption and distributes uniformly in the blood stream by chemically binding to albumin. With the help of EB, complete and continuous microvascular networks—especially capillaries—are imaged. The diffusion dynamics of EB leaving the blood stream and the clearance dynamics of the EB-albumin complex are also quantitatively investigated.

Select this Article		Experimental investigation of evanescence-based infrared biodetection technique for micro-total-analysis systems Arvind Chandrasekaran and Muthukumaran Packirisamy J. Biomed. Opt. 14, 054050 (Oct 30, 2009); http://dx.doi.org/10.1117/1.3210766 Online Publication Date: Oct 30, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract The advent of microoptoelectromechanical systems (MOEMS) and its integration with other technologies such as microfluidics, microthermal, immunoproteomics, etc. has led to the concept of an integrated micro-total-analysis systems (μTAS) or Lab-on-a-Chip for chemical and biological applications. Recently, research and development of μTAS have attained a significant growth rate over several biodetection sciences, in situ medical diagnoses, and point-of-care testing applications. However, it is essential to develop suitable biophysical label-free detection methods for the success, reliability, and ease of use of the μTAS. We proposed an infrared (IR)-based evanescence wave detection system on the silicon-on-insulator platform for biodetection with μTAS. The system operates on the principle of bio-optical interaction that occurs due to the evanescence of light from the waveguide device. The feasibility of biodetection has been experimentally investigated by the detection of horse radish peroxidase upon its reaction with hydrogen peroxide.

Select this Article OPEN ACCESS		Quantitative optical spectroscopy can identify long-term local tumor control in irradiated murine head and neck xenografts Karthik Vishwanath, Daniel Klein, Kevin Chang, Thies Schroeder, Mark W. Dewhirst, and Nimmi Ramanujam J. Biomed. Opt. 14, 054051 (Oct 23, 2009); http://dx.doi.org/10.1117/1.3251013 \| Cited 5 times Online Publication Date: Oct 23, 2009 Full Text: Read Online (HTML) \| Download PDF
	+ -	Show Abstract Noninvasive and longitudinal monitoring of tumor oxygenation status using quantitative diffuse reflectance spectroscopy is used to test whether a final treatment outcome could be estimated from early optical signatures in a murine model of head and neck cancer when treated with radiation. Implanted tumors in the flank of 23 nude mice are exposed to 39 Gy of radiation, while 11 animals exposed to sham irradiation serve as controls. Diffuse optical reflectance is measured from the tumors at baseline (prior to irradiation) and then serially until 17 days posttreatment. The fastest and greatest increase in baseline-corrected blood oxygen saturation levels are observed from the animals that show complete tumor regression with no recurrence 90 days postirradiation, relative to both untreated and treated animals with local recurrences. These increases in saturation are observed starting 5 days posttreatment and last up to 17 days posttreatment. This preclinical study demonstrates that diffuse reflectance spectroscopy could provide a practical method far more effective than the growth delay assay to prognosticate treatment outcome in solid tumors and may hold significant translational promise.

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Errata

Select this Article OPEN ACCESS		Errata: Simultaneous photoacoustic imaging technique using an acoustic imaging lens Xian Chen, Zhilie Tang, Yongheng He, Haifeng Liu, Yadong Wei, and Yongbo Wu J. Biomed. Opt. 14, 059801 (Oct 07, 2009); http://dx.doi.org/10.1117/1.3223339 \| Cited 1 time Online Publication Date: Oct 07, 2009 Full Text: Read Online (HTML) \| Download PDF
		Abstract Unavailable

Select this Article OPEN ACCESS		Errata: Confocal reflectance and two-photon microscopy studies of a songbird skull for preparation of transcranial imaging Darine Abi-Haidar and Thomas Olivier J. Biomed. Opt. 14, 059802 (Sep 22, 2009); http://dx.doi.org/10.1117/1.3223351 Online Publication Date: Sep 22, 2009 Full Text: Read Online (HTML) \| Download PDF
		Abstract Unavailable

Select this Article OPEN ACCESS		Errata: Visualization of hepatobiliary excretory function by intravital multiphoton microscopy Yuan Liu, Hsiao-Ching Chen, Shu-Mei Yang, Tzu-Lin Sun, Wen Lo, Ling-Ling Chiou, Guan Tarn Huang, Chen-Yuan Dong, and Hsuan-Shu Lee J. Biomed. Opt. 14, 059803 (Oct 20, 2009); http://dx.doi.org/10.1117/1.3249658 Online Publication Date: Oct 20, 2009 Full Text: Read Online (HTML) \| Download PDF
		Abstract Unavailable

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