For the first time, the use of fiber-optic colorDoppler optical coherence tomography (CDOCT) to mapin vivo the three-dimensional (3-D) vascular network of airwaysegments in human lungs is demonstrated. Visualizingthe 3-D vascular network in the lungs may provide newopportunities for detecting and monitoring lung diseasessuch as asthma, chronic obstructive pulmonary disease, andlung cancer. Our CDOCT instrument employs a rotary fiberopticprobe that provides simultaneous two-dimensional (2-D) real-time structural optical coherence tomography (OCT)and CDOCT imaging at frame rates up to 12.5 frames persecond. Controlled pullback of the probe allows 3-D vascularmapping in airway segments up to 50 mm in length in asingle acquisition. We demonstrate the ability of CDOCT tomap both small and large vessels. In one example, CDOCTimaging allows assignment of a feature in the structural OCTimage as a large (∼1 mm diameter) blood vessel. In a secondexample, a smaller vessel (∼80 μm diameter) that is indistinguishablein the structural OCT image is fully visualized in3-D using CDOCT.

Miniature endoscopic probes offer a solution for deep brain imaging by overcoming the limited depth of intravital microscopy. We describe a small-diameter (350 μm) graded-index optical probe with a side-view design for in vivo cellular imaging of the mammalian brain. The side-view probe provides a unique view of the vertical network of neurons and penetrating blood vessels. At a given insertion site, the translational and rotational scanning of the probe provides access to a large tissue area (<1  mm ² ) across the cortex, hippocampus, thalamus, and hypothalamus.

The efficacy of radiation therapy depends strongly on tumor oxygenation during irradiation. However, current techniques to measure this parameter in vivo do not facilitate routine monitoring in patients. Herein, we demonstrate a noninvasive method for tomographic imaging of oxygen partial pressure (pO₂ ) in deep tissue using the phosphorescence decay of an oxygen-sensitive probe excited by Čerenkov radiation induced by external beam radiotherapy. Tissue-simulating scattering phantoms (60 mm diameter with a 20 mm anomaly) containing platinum(II)-G4 (PtG4), a dendritic porphyrin-based phosphor, whose phosphorescence is quenched in the presence of oxygen, were irradiated with a clinical linear accelerator. The emitted phosphorescence was measured at various positions on the phantom boundary using a spectrograph coupled to an intensified charge-coupled device (ICCD). At each position, PtG4 phosphorescence decay curves were measured by synchronizing the ICCD to the linear accelerator pulses. Tomographic images of phosphorescence yield and lifetime were recovered for phantoms with homogenous PtG4 concentrations and heterogeneous pO₂ . Since PtG4 lifetime is strongly and predictably dependent on pO ₂ through the Stern-Volmer relationship, tomographic images of pO ₂ were also reported, and showed excellent agreement with independent oxygenation measurements. Translating this approach to the clinic could facilitate direct sensing of pO₂ during radiotherapy.

Multispectral imaging has shown promise in subcutaneous vein detection and localization in human subjects. While many limitations of single-wavelength methods are addressed in multispectral vein detection methods, their performance is still limited by artifacts arising from background skin reflectance and optimality of postprocessing algorithms. We propose a background removal technique that enhances the contrast and performance of multispectral vein detection. We use images acquired at visible wavelengths as reference for removing skin reflectance background from subcutaneous structures in near-infrared images. Results are validated by experiments on human subjects.

The spindle plays a crucial role in normal chromosome alignment and segregation during meiosis and mitosis. Studying spindles in living cells noninvasively is of great value in assisted reproduction technology (ART). Here, we present a novel spindle imaging methodology, full-field optical coherence tomography (FF-OCT). Without any dye labeling and fixation, we demonstrate the first successful application of FF-OCT to noninvasive three-dimensional (3-D) live imaging of the meiotic spindles within the mouse living oocytes at metaphase II as well as the mitotic spindles in the living zygotes at metaphase and telophase. By post-processing of the 3-D dataset obtained with FF-OCT, the important morphological and spatial parameters of the spindles, such as short and long axes, spatial localization, and the angle of meiotic spindle deviation from the first polar body in the oocyte were precisely measured with the spatial resolution of 0.7 μm. Our results reveal the potential of FF-OCT as an imaging tool capable of noninvasive 3-D live morphological analysis for spindles, which might be useful to ART related procedures and many other spindle related studies.

A recently reported micro-optical sectioning tomography system has great potential to draw the neuronal circuits of large brain volume with submicron resolution by combining fine mechanic sectioning with simultaneous optical imaging. However, sectioning the fluorescence sample sometimes induces tears between adjacent tiles and causes difficulties in continuous fiber tracing from fluorescence imaging. A confocal detection to recover the interruptions of the nerve fiber is introduced. With a 50-μm-width confocal slit, the signal-to-background ratio is increased 16- to 49-fold more than that without the slit, which effectively improves the detectability of the signal in the interruptions and enables continuous tracing of the neuronal circuits.

High-resolution mapping of microvasculature has been applied to diverse body systems, including the retinal and choroidal vasculature, cardiac vasculature, the central nervous system, and various tumor models. Many imaging techniques have been developed to address specific research questions, and each has its own merits and drawbacks. Understanding, optimization, and proper implementation of these imaging techniques can significantly improve the data obtained along the spectrum of unique research projects to obtain diagnostic clinical information. We describe the recently developed algorithms and applications of two general classes of microvascular imaging techniques: speckle-variance and phase-variance optical coherence tomography (OCT). We compare and contrast their performance with Doppler OCT and optical microangiography. In addition, we highlight ongoing work in the development of variance-based techniques to further refine the characterization of microvascular networks.

A general survey is provided on the capability of Monte Carlo (MC) modeling in tissue optics while paying special attention to the recent progress in the development of methods for speeding up MC simulations. The principles of MC modeling for the simulation of light transport in tissues, which includes the general procedure of tracking an individual photon packet, common light–tissue interactions that can be simulated, frequently used tissue models, common contact/noncontact illumination and detection setups, and the treatment of time-resolved and frequency-domain optical measurements, are briefly described to help interested readers achieve a quick start. Following that, a variety of methods for speeding up MC simulations, which includes scaling methods, perturbation methods, hybrid methods, variance reduction techniques, parallel computation, and special methods for fluorescence simulations, as well as their respective advantages and disadvantages are discussed. Then the applications of MC methods in tissue optics, laser Doppler flowmetry, photodynamic therapy, optical coherence tomography, and diffuse optical tomography are briefly surveyed. Finally, the potential directions for the future development of the MC method in tissue optics are discussed.

As optical fibers revolutionize the way data is carried in telecommunications, the same is happening in the world of sensing. Fiber-optic sensors (FOS) rely on the principle of changing the properties of light that propagate in the fiber due to the effect of a specific physical or chemical parameter. We demonstrate the potentialities of this sensing concept to assess pressure in biomedical and biomechanical applications. FOSs are introduced after an overview of conventional sensors that are being used in the field. Pointing out their limitations, particularly as minimally invasive sensors, is also the starting point to argue FOSs are an alternative or a substitution technology. Even so, this technology will be more or less effective depending on the efforts to present more affordable turnkey solutions and peer-reviewed papers reporting in vivo experiments and clinical trials.

The invariant imbedding T-matrix method (II-TM) is employed to simulate the optical properties of normal biconcave and deformed red blood cells (RBCs). The phase matrix elements of a RBC model computed with the II-TM are compared with their counterparts computed with the discrete-dipole approximation (DDA) method. As expected, the DDA results approach the II-TM results with an increase in the number of dipoles per incident wavelength. Computationally, the II-TM is faster than the DDA when multiple RBC orientations are considered. For a single orientation, the DDA is comparable with or even faster than the II-TM because the DDA efficiently converges for optically soft particles; however, the DDA method demands significantly more computer memory than the II-TM. After the applicability of the II-TM is numerically confirmed, a comparison is conducted of the optical properties of oxygenated and deoxygenated RBCs and of normal and deformed RBCs. The spectral variations of RBCs’ optical properties are investigated in the wavelength range from 0.25 to 1.0 μm. Furthermore, the statistically averaged phase matrix of spheres and biconcave RBCs are compared. Conducted numerical simulations suggest the applicability of the II-TM for the inverse light scattering analysis and radiative transfer simulations in blood.

Laser microdissection by pulsing ultraviolet laser allows the isolation and recultivation of live cells based on morphological features or/and fluorescent labelling from adherent cell cultures. Previous investigations described only the use of the laser microdissection and pressure catapulting (LMPC) for live cell isolation. But LMPC requires complex manipulations and some skill. Furthermore, single-cell cloning using laser microdissection has not yet been demonstrated. The first evidence of successful application of laser microdissection with gravity transfer (LMDGT) for capturing and recultivation of live cells is presented. A new strategy for LMDGT is presented because of the failure to reproduce the manufacturer’s protocol. Using the new strategy, successful capturing and recultivation of circle-shaped samples from confluent monolayer of HeLa cells was demonstrated. It was found that LMDGT is easier than LMPC because it doesn’t require personal participation of investigator in transferring of isolated samples to final culture dishes. Moreover, for the first time, the generation of clonal colonies from single live cells isolated by laser microdissection was demonstrated. Data obtained in this study confirm that LMDGT is a reliable and high-yield method allowing isolation and expansion of both cell clusters and single cells from adherent cell cultures.

Raman active nano-complex agents based on single-walled carbon nanotubes (SWCNTs) are prepared and used for the swift and specific detection of breast cancer cells. SWCNTs are functionalized to bond covalently with the anti-epithelial cell adhesion molecule (anti-EpCAM) antibody, which is specific to the highly expressed EpCAM antigen on the surface of breast cancer cells (MCF-7), but not on normal cells. The Raman nano-complexes demonstrate excellent ability to detect in vitro single breast cancer cells (MCF-7) and discriminate between them and normal fibroblast cells during the first 30 min of the targeting process. Raman linearity scanning is collected from a monolayer cell mixture, including both cancer cells and normal cells incubated with anti-EpCAM-SWCNTs, using a 633-nm laser excitation. The results shows that the Raman signal collected from targeted MCF-7 cells is extremely high, whereas there is little signal from the normal cells.

Photodynamic therapy has been considered a potential antimicrobial modality against oral infections, including dental caries. A model to estimate the penetration of both photosensitizers and light through human dentin, a factor of interest in photodynamic therapy, is proposed. The photoacoustic spectroscopy technique was used to evaluate in vitro dentin permeability of three different photosensitizers. Using the dentin optical absorption and scattering coefficients, it was possible to propose a semi-quantitative model predicting both photosensitizer and light doses within dentin. The graphic illustrations obtained provided guidelines that may be useful in photodynamic therapy protocols used as antimicrobial tools in caries lesions.

There is potential for Raman spectroscopy (RS) to complement tools for bone diagnosis due to its ability to assess compositional and organizational characteristics of both collagen and mineral. To aid this potential, the present study assessed specificity of RS peaks to the composition of bone, a birefringent material, for different degrees of instrument polarization. Specifically, relative changes in peaks were quantified as the incident light rotated relative to the orientation of osteonal and interstitial tissue, acquired from cadaveric femurs. In a highly polarized instrument (10 ⁶ ∶1 extinction ratio), the most prominent mineral peak (ν1 Phosphate at 961  cm ⁻¹ ) displayed phase similarity with the Proline peak at 856  cm ^−1[/sup] . This sensitivity to relative orientation between bone and light observed in the highly polarized regime persisted for certain sensitive peaks (e.g., Amide I at 1666  cm ⁻¹ ) in unaltered instrumentation (200∶1 extinction ratio). Though Proline intensity changed with bone rotation, the phase of Proline matched that of ν1 Phosphate. Moreover, when mapping ν1 Phosphate/Proline across osteonal-interstitial borders, the mineralization difference between the tissue types was evident whether using a 20x or 50x objectives. Thus, the polarization bias inherent in commercial RS systems does not preclude the assessment of bone composition when using phase-matched peaks.

In recent years, advances in technology are propelling the field of oral and maxillofacial surgery into new realms. With a relatively thin alveolar mucosa overlying the underlying bone, significant diagnostic and therapeutic advantages are present; however, there remains an enormous gap between advancements in physics, in particular optics, and oral and maxillofacial surgery. Improvements in diagnosis, classification, and treatment of the various bone pathologies are still being sought after as advancements in technology continue to progress. Combining the clinical, histological, and pathological characteristics with these advancements, patients with debilitating pathologies may have more promising treatment options and prognosis. Defects in the facial bones, particularly in the jaws, may be due to a number of reasons: pathology, trauma, infections, congenital deformities, or simply due to atrophy. Bone grafting is commonly employed to correct such defects, and allows new bone formation through tissue regeneration. Growing use of dental implants has focused attention on osseointegration and its process. Osseointegration refers to the actual process of the direct contact between bone and implant, without an intervening soft tissue layer. The theories proposed regarding this process are many, yet a clear, unified stance on the actual process and its mechanisms has not emerged. Further investigation using optical probes could provide that unifying answer. The primary goal of this manuscript is to introduce pioneers in the field of optics to the field of oral and maxillofacial surgery. With a brief introduction into the procedures and techniques, we are hopeful to bridge the ever-widening gap between the clinical science and the basic sciences.

In continuous-wave near-infrared spectroscopy (CW-NIRS), changes in the concentration of oxyhemoglobin and deoxyhemoglobin can be calculated by solving a set of linear equations from the modified Beer-Lambert Law. Cross-talk error in the calculated hemodynamics can arise from inaccurate knowledge of the wavelength-dependent differential path length factor (DPF). We apply the extended Kalman filter (EKF) with a dynamical systems model to calculate relative concentration changes in oxy- and deoxyhemoglobin while simultaneously estimating relative changes in DPF. Results from simulated and experimental CW-NIRS data are compared with results from a weighted least squares (WLSQ) method. The EKF method was found to effectively correct for artificially introduced errors in DPF and to reduce the cross-talk error in simulation. With experimental CW-NIRS data, the hemodynamic estimates from EKF differ significantly from the WLSQ (p<0.001 ). The cross-correlations among residuals at different wavelengths were found to be significantly reduced by the EKF method compared to WLSQ in three physiologically relevant spectral bands 0.04 to 0.15 Hz, 0.15 to 0.4 Hz and 0.4 to 2.0 Hz (p<0.001 ). This observed reduction in residual cross-correlation is consistent with reduced cross-talk error in the hemodynamic estimates from the proposed EKF method.

Pancreatic stellate cells (PSCs) and other pancreatic components that play a critical role in exocrine pancreatic diseases are generally identified separately by conventional studies, which provide indirect links between these components. Here, nonlinear optical microscopy was evaluated for simultaneous characterization of these components within a three-dimensional (3-D) tissue environment, primarily based on multichannel detection of intrinsic optical emissions and cell morphology. Fresh rat pancreatic tissues harvested at 1 day, 7 days, and 28 days after induction of chronic pancreatitis were imaged, respectively. PSCs, inflammatory cells, blood vessels, and collagen fibers were identified simultaneously. The PSCs at day 1 of chronic pancreatitis showed significant enlargement compared with those in normal pancreas (p<0.001 , analysis of variance linear contrast; n=8 for each group). Pathological events relating to these components were observed, including presence of inflammatory cells, deposited collagen, and phenotype conversion of PSCs. We demonstrate that label-free nonlinear optical microscopy is an efficient tool for dissecting PSCs and other pancreatic components coincidently within 3-D pancreatic tissues. It is a prospect for intravital observation of dynamic events under natural physiological conditions, and might help uncover the key mechanisms of exocrine pancreatic diseases, leading to more effective treatments.

Optical coherence tomography (OCT) images can provide quantitative measurements of the eye’s entire anterior segment. A new technique founded on a newly proposed level set-based shape prior segmentation model has been developed for automatic segmentation of the cornea’s anterior and posterior boundaries. This technique comprises three major steps: removal of regions containing irrelevant structures and artifacts, estimation of the cornea’s location using a thresholding technique, and application of the new level set-based shape prior segmentation model to improve segmentation. The performance of our technique is compared to previously developed methods for analysis of the cornea in 33 OCT images of normal eyes, whereby manual annotations are used as a reference standard. The new technique achieves much improved concordance than previous methods, with a mean Dice’s similarity coefficient of <0.92 . This demonstrates the technique’s potential to provide accurate and reliable measurements of the anterior segment geometry, which is important for many applications, including the construction of representative numerical simulations of the eye’s mechanical behavior.

Activatable photoacoustic probes efficiently combine the high spatial resolution and penetration depth of ultrasound with the high optical contrast and versatility of molecular imaging agents. Our approach is based on photoacoustic probing of the excited-state lifetime of methylene blue (MB), a fluorophore widely used in clinical therapeutic and diagnostic applications. Upon aggregation, static quenching between the bound molecules dramatically shortens their lifetime by three orders of magnitude. We present preliminary results demonstrating the ability of photoacoustic imaging to probe the lifetime contrast between monomers and dimers with high sensitivity in cylindrical phantoms. Gradual dimerization enhancement, driven by the addition of increasing concentrations of sodium sulfate to a MB solution, showed that lifetime-based photoacoustic probing decreases linearly with monomer concentration. Similarly, the addition of 4 mM sodium dodecyl sulfate, a concentration that amplifies MB aggregation and reduces the monomer concentration by more than 20-fold, led to a signal decrease of more than 20 dB compared to a solution free of surfactant. These results suggest that photoacoustic imaging can be used to selectively detect the presence of monomers. We conclude by discussing the implementation of the monomer–dimer contrast mechanism for the development of an enzyme-specific activatable probe.

A swept-source dual-wavelength photothermal (DWP) optical coherence tomography (OCT) system is demonstrated for quantitative imaging of microvasculature oxygen saturation. DWP-OCT is capable of recording three-dimensional images of tissue and depth-resolved phase variation in response to photothermal excitation. A 1,064-nm OCT probe and 770-nm and 800-nm photothermal excitation beams are combined in a single-mode optical fiber to measure microvasculature hemoglobin oxygen saturation (SO₂ ) levels in phantom blood vessels with a range of blood flow speeds (0 to 17  mm/s ). A 50-μm-diameter blood vessel phantom is imaged, and SO₂ levels are measured using DWP-OCT and compared with values provided by a commercial oximeter at various blood oxygen concentrations. The influences of blood flow speed and mechanisms of SNR phase degradation on the accuracy of SO ₂ measurement are identified and investigated.

The ability to visualize early stage lung cancer is important in the study of biomarkers and targeting agents that could lead to earlier diagnosis. The recent development of hybrid free-space 360-deg fluorescence molecular tomography (FMT) and x-ray computed tomography (XCT) imaging yields a superior optical imaging modality for three-dimensional small animal fluorescence imaging over stand-alone optical systems. Imaging accuracy was improved by using XCT information in the fluorescence reconstruction method. Despite this progress, the detection sensitivity of targeted fluorescence agents remains limited by nonspecific background accumulation of the fluorochrome employed, which complicates early detection of murine cancers. Therefore we examine whether x-ray CT information and bulk fluorescence detection can be combined to increase detection sensitivity. Correspondingly, we research the performance of a data-driven fluorescence background estimator employed for subtraction of background fluorescence from acquisition data. Using mice containing known fluorochromes ex vivo, we demonstrate the reduction of background signals from reconstructed images and sensitivity improvements. Finally, by applying the method to in vivo data from K-ras transgenic mice developing lung cancer, we find small tumors at an early stage compared with reconstructions performed using raw data. We conclude with the benefits of employing fluorescence subtraction in hybrid FMT-XCT for early detection studies.

Small animal models of retinal diseases are important to vision research, and noninvasive high resolution in vivo rodent retinal imaging is becoming an increasingly important tool used in this field. We present a custom Fourier domain optical coherence tomography (FD-OCT) instrument for high resolution imaging of mouse retina. In order to overcome aberrations in the mouse eye, we incorporated a commercial adaptive optics system into the sample arm of the refractive FD-OCT system. Additionally, a commercially available refraction canceling lens was used to reduce lower order aberrations and specular back-reflection from the cornea. Performance of the adaptive optics (AO) system for correcting residual wavefront aberration in the mice eyes is presented. Results of AO FD-OCT images of mouse retina acquired in vivo with and without AO correction are shown as well.

The ability to image metallic implants is important for medical applications ranging from diagnosis to therapy. Photoacoustic (PA) imaging has been recently pursued as a means to localize metallic implants in soft tissue. The work presented herein investigates different mechanisms to modulate the PA signal generated by macroscopic metallic surfaces. Wires of five different metals are tested to simulate medical implants/tools, while surface roughness is altered or physical vapor deposition (PVD) coatings are added to change the wires’ overall optical absorption. PA imaging data of the wires are acquired at 970 nm. Results indicate that PA signal generation predominately occurs in a wire’s metallic surface and not its aqueous surroundings. PA signal generation is similar for all metals tested, while addition of PVD coatings offers significant modulations (i.e., 4-dB enhancement and 26-dB reduction achieved) in PA signal generation. Results also suggest that PA signal increases with increasing surface roughness. Different coating and roughness schemes are then successfully utilized to generate spatial PA signal patterns. This work demonstrates the potential of surface modifications to enhance or reduce PA signal generation to permit improved PA imaging of implants/tools (i.e., providing location/orientation information) or to allow PA imaging of surrounding tissue.

Intravascular near-infrared fluorescence (NIRF) imaging offers a new approach for characterizing atherosclerotic plaque, but random catheter positioning within the vessel lumen results in variable light attenuation and can yield inaccurate measurements. We hypothesized that NIRF measurements could be corrected for variable light attenuation through blood by tracking the location of the NIRF catheter with intravascular ultrasound (IVUS). In this study, a combined NIRF-IVUS catheter was designed to acquire coregistered NIRF and IVUS data, an automated image processing algorithm was developed to measure catheter-to-vessel wall distances, and depth-dependent attenuation of the fluorescent signal was corrected by an analytical light propagation model. Performance of the catheter sensing distance correction method was evaluated in coronary artery phantoms and ex vivo arteries. The correction method produced NIRF estimates of fluorophore concentrations, in coronary artery phantoms, with an average root mean square error of 17.5%. In addition, the correction method resulted in a statistically significant improvement in correlation between spatially resolved NIRF measurements and known fluorophore spatial distributions in ex vivo arteries (from r=0.24 to 0.69, p<0.01 , n=6 ). This work demonstrates that catheter-to-vessel wall distances, measured from IVUS images, can be employed to compensate for inaccuracies caused by variable intravascular NIRF sensing distances.

Confocal Raman microscopy is a noninvasive, label-free imaging technique used to study apoptosis of live MCF-7 cells. The images are based on Raman spectra of cells components, and their apoptosis is monitored through diffusion of cytochrome c in cytoplasm. K-mean clustering is used to identify mitochondria in cells, and correlation analysis provides the cytochrome c distribution inside the cells. Our results demonstrate that incubation of cells for 3 h with 10 μM of paclitaxel does not induce apoptosis in MCF-7 cells. On the contrary, incubation for 30 min at a higher concentration (100 μM) of paclitaxel induces gradual release of the cytochrome c into the cytoplasm, indicating cell apoptosis via a caspase independent pathway.

Optical techniques such as speckle pattern interferometry are well known in the nondestructive testing measurement community. They can be used, for instance, as a predictor of the mechanical behavior of a sample under study. However, in almost all circumstances, a mathematical model has to be applied in order to make sense of these measurements. This is a critical issue when an organic sample is studied, mainly due to its complex deformation response. A good example of this is observed in the birds’ feathers. They have extraordinary mechanical and aerodynamic properties thanks to their stiffness and lightness. A couple of live birds are safely situated in front of an out-of-plane sensitive digital holographic interferometer (DHI), an optical system capable of recovering the optical phase in this type of nonrepeatable or unpredictable experiment. In order to recover the backscattering signal and its interferometric response, several images are recorded from different sections of the plumage. Displacement maps are obtained from what is, as far as is known, the first time that full field microdisplacement maps are presented over a hummingbird and a parakeet plumage.

Optical mammography is a functional imaging technique that uses near-infrared light to produce three-dimensional breast images of tissue oxygen saturation and hemoglobin concentration. It has been used to monitor the response to neoadjuvant chemotherapy in breast cancer patients. We present the first results on monitoring tumor response to hormone therapy using optical mammography. We present three case studies from postmenopausal women treated with neoadjuvant hormone therapy for locally advanced breast cancer. The women were scanned before starting treatment, once during treatment, and then before surgery. Changes in physiological and optical properties within the tumor and in the rest of the breast were evaluated. At the time of surgery, two patients partially responded to treatment and one did not respond. The patients that partially responded on ultrasound revealed a corresponding recovery to normal in the hemoglobin concentration images, whereas the nonresponder indicated an increase in hemoglobin concentration in the tumor compared to her pretreatment images. These case studies suggest that optical imaging of the breast during neoadjuvant hormone treatment can provide potentially valuable information, and that physiological changes within the tumor can be seen in response to treatment.

Fluorescence molecular tomography (FMT) is an important imaging technique of optical imaging. The major challenge of the reconstruction method for FMT is the ill-posed and underdetermined nature of the inverse problem. In past years, various regularization methods have been employed for fluorescence target reconstruction. A comparative study between the reconstruction algorithms based on l ₁ -norm and l ₂ -norm for two imaging models of FMT is presented. The first imaging model is adopted by most researchers, where the fluorescent target is of small size to mimic small tissue with fluorescent substance, as demonstrated by the early detection of a tumor. The second model is the reconstruction of distribution of the fluorescent substance in organs, which is essential to drug pharmacokinetics. Apart from numerical experiments, in vivo experiments were conducted on a dual-modality FMT/micro-computed tomography imaging system. The experimental results indicated that l ₁norm regularization is more suitable for reconstructing the small fluorescent target, while l ₂-norm regularization performs better for the reconstruction of the distribution of fluorescent substance.

We report on a structured light-scanning system, the OGX|4DSCANNER, capable of capturing the surface of a human body with 2 mm spatial resolution at a 60 Hz frame-rate. The performance of modeling the human lower body dynamics is evaluated by comparing the system with the current gold standard, i.e., the VICON system. The VICON system relies on the application of reflective markers on a person’s body and tracking their positions in three-dimensional space using multiple cameras [optical motion capture (OMC)]. For the purpose of validation of the 4DSCANNER, a set of “virtual” markers was extracted from the measured surface. A set of musculoskeletal models was built for three subjects based on the trajectories of real and virtual markers. Next, the corresponding models were compared in terms of joint angles, joint moments, and activity of a number of major lower body muscles. Analyses showed a good overall agreement of the modeling outcome. We conclude that the 4DSCANNER within its limitations has the potential to be used in clinical gait analysis instead of optical marker-based systems. The advantage of the 4DSCANNER over OMC solutions is that it does not burden patients with time-consuming marker application. This study demonstrates the versatility of this measurement technique.

Diffuse correlation spectroscopy (DCS) measurements in vivo recorded from rabbits’ ocular fundus are presented. Despite the complexity of these ocular tissues, we provide a clear and simple demonstration of the DCS abilities to analyze variations in physiological quantities of clinical interest. Indeed, the reported experimental activities demonstrate that DCS can reveal both choroidal-flow and temperature variations and detect nano- and micro-aggregates in ocular fundus circulation. Such abilities can be of great interest both in fundamental research and practical clinical applications. The proposed measuring system can be useful in: (a) monitoring choroidal blood flow variations, (b) determining the end-point for photo-dynamic therapy and transpupillary thermo therapy and, (c) managing the dye injection and determining an end-point for dye-enhanced photothrombosis. Moreover, it could allow both diagnoses when the presence of nano- and micro-aggregates is related to specific diseases and verifying the effects of nanoparticle injection in nanomedicine. Even though the reported results demonstrate the applicability of DCS to investigate ocular fundus, a detailed and accurate investigation of the limits of detection is beyond the scope of this article.

We report an overlooked source of artifacts for clinical specimens, where unexpected and normally negligible contaminants can skew the interpretation of results. During an ongoing study of bone fragments from diabetic osteomyelitis, strong Raman signatures were found, which did not correspond with normal bone mineral or matrix. In a bone biopsy from the calcaneus of a patient affected by diabetic osteomyelitis, Raman microspectroscopic analysis revealed regions with both abnormal mineral and degraded collagen in addition to normal bone. Additional bands indicated a pathological material. Stenotrophomonas maltophilia was identified in the wound culture by independent microbiologic examination. We initially assigned the unusual bands to xanthomonadin, a bacterial pigment from S. maltophilia. However, the same bands were also found more than a year later on a second specimen that had been noticeably contaminated with pathology marking dye. Drop deposition/Raman spectroscopy of commonly used pathology dyes revealed that a blue tissue-marking dye was responsible for the unusual bands in both specimens, even in the first specimen where there was no visible evidence of contamination.

An appropriate method for spectrum extraction for better signal-to-noise ratio (SNR) and lower computational cost is essential in noninvasive detection. We have first studied two existing extraction methods for dynamic spectrum (DS) comparatively: frequency domain analysis and single trial estimation; after analyzing the advantages and disadvantages theoretically, a new method based on a fast digital lock-in amplifier (FDLIA) was developed to overcome the limitations of these two existing methods. The feasibility of the new method was verified by experiments and the results demonstrated that the FDLIA method based on DS had greatly simplified the computation of frequency domain analysis without the method error; moreover, the continuous signal was cut into several short segments in FDLIA and the gross errors from an episode of pulse wave were eliminated, and thus SNR improved. Therefore, the FDLIA method utilizing the advantages of both existing methods can be effectively realized in a general embedded system in real time for its simple algorithm.

We describe a novel sensing approach based on a functionalized microstructured optical fiber-Bragg grating for specific DNA target sequences detection. The inner surface of a microstructured fiber, where a Bragg grating was previously inscribed, has been functionalized by covalent linking of a peptide nucleic acid probe targeting a DNA sequence bearing a single point mutation implicated in cystic fibrosis (CF) disease. A solution of an oligonucleotide (ON) corresponding to a tract of the CF gene containing the mutated DNA has been infiltrated inside the fiber capillaries and allowed to hybridize to the fiber surface according to the Watson-Crick pairing. In order to achieve signal amplification, ON-functionalized gold nanoparticles were then infiltrated and used in a sandwich-like assay. Experimental measurements show a clear shift of the reflected high order mode of a Bragg grating for a 100 nM DNA solution, and fluorescence measurements have confirmed the successful hybridization. Several experiments have been carried out on the same fiber using the identical concentration, showing the same modulation trend, suggesting the possibility of the reuse of the sensor. Measurements have also been made using a 100 nM mismatched DNA solution, containing a single nucleotide mutation and corresponding to the wild-type gene, and the results demonstrate the high selectivity of the sensor.

We determined the optical properties of turbid media from simulated spatially resolved reflectance (SRR) curves using an artificial neural network (ANN). In order to improve the performance of our method, multiple ANNs were applied for this problem. First, Monte Carlo (MC) simulations were performed using random optical properties which are relevant for biological tissue. For a better performance of the ANN in respect of SRR measurements, the exact setup geometry was taken into account for the MC simulations. Second, the performed simulations were classified into different categories according to their shape. Third, multiple ANNs which were adjusted to these categories, were used to solve the inverse problem, i.e., the determination of the optical properties from SRR curves. Finally, these ANNs were applied to determine the optical properties of simulated SRR curves out of the range 0.5  mm ⁻¹ <μ ′ s <5  mm⁻¹ and 0.0001  mm⁻¹ <μ a <1  mm⁻¹ . The average relative error was 2.9% and 6.1% for the reduced scattering coefficient μ ′ _s and for the absorption coefficient μ_a , respectively.

We present a fiber-optic sensor for monitoring respiration and heart activity designed to operate in the magnetic resonance imaging (MRI) environment. The sensor employs a Plexiglas springboard, which converts movements of the patient’s body lying on the board (i.e., lung- and heart-induced vibrations) to strain, where a fiber Bragg grating attached to the board is used to measure this strain. Experimental studies are carried out during thoracic spine MRI examinations. The presence of the metal-free sensor construction in the MRI environment does not pose a threat to the patient and has no influence over the quality of imaging, and the signal is identical to that obtained without any electromagnetic interference. The results show that the sensor is able to accurately reflect the ballistocardiographic signal, enabling determinations of the respiration rate (RR) and heart rate (HR). The data delivered by the sensor are normally distributed on the Bland–Altman plot for the characteristic point determination and exhibit clear dependence on the RR and HR values for the RR and HR determinations, respectively. Measurement accuracies are better than 7% of the average values, and thus, with further development, the sensor will be implemented in routine MRI examinations.

Peripheral artery disease (PAD) is a common condition with high morbidity. While measurement of tissue oxygen saturation (S _t O ₂ ) has been demonstrated, this is the first study to assess both S _t O ₂ and relative blood flow (rBF) in the extremities of PAD patients. Diffuse optics is employed to measure hemodynamic response to treadmill and pedal exercises in 31 healthy controls and 26 patients. For S _t O ₂ , mild and moderate/severe PAD groups show pronounced differences compared with controls. Pre-exercise mean S _t O ₂ is lower in PAD groups by 9.3% to 10.6% compared with means of 63.5% to 66.2% in controls. For pedal, relative rate of return of S _t O ₂ to baseline is more rapid in controls (p<0.05 ). Patterns of rBF also differ among groups. After both exercises, rBF tend to occur at depressed levels among severe PAD patients compared with healthy (p<0.05 ); post-treadmill, rBF tend to occur at elevated levels among healthy compared with severe PAD patients (p<0.05 ). Additionally, relative rate of return to baseline S _t O ₂ is more rapid among subjects with reduced levels of depression in rBF (p=0.041 ), even after adjustment for ankle brachial index. This suggests a physiologic connection between rBF and oxygenation that can be measured using diffuse optics, and potentially employed as an evaluative tool in further studies.

We describe an approach for estimating human skin parameters, such as melanosome concentration, collagen concentration, oxygen saturation, and blood volume, using hyperspectral radiometric measurements (signatures) obtained from in vivo skin. We use a computational model based on Kubelka-Munk theory and the Fresnel equations. This model forward maps the skin parameters to a corresponding multiband reflectance spectra. Machine-learning-based regression is used to generate the inverse map, and hence estimate skin parameters from hyperspectral signatures. We test our methods using synthetic and in vivo skin signatures obtained in the visible through the short wave infrared domains from 24 patients of both genders and Caucasian, Asian, and African American ethnicities. Performance validation shows promising results: good agreement with the ground truth and well-established physiological precepts. These methods have potential use in the characterization of skin abnormalities and in minimally-invasive prescreening of malignant skin cancers.

Suture ligation of blood vessels during surgery can be time-consuming and skill-intensive. Energy-based, electrosurgical, and ultrasonic devices have recently replaced the use of sutures and mechanical clips (which leave foreign objects in the body) for many surgical procedures, providing rapid hemostasis during surgery. However, these devices have the potential to create an undesirably large collateral zone of thermal damage and tissue necrosis. We explore an alternative energy-based technology, infrared lasers, for rapid and precise thermal coagulation and fusion of the blood vessel walls. Seven near-infrared lasers (808, 980, 1075, 1470, 1550, 1850 to 1880, and 1908 nm) were tested during preliminary tissue studies. Studies were performed using fresh porcine renal vessels, ex vivo, with native diameters of 1 to 6 mm, and vessel walls flattened to a total thickness of 0.4 mm. A linear beam profile was applied normal to the vessel for narrow, full-width thermal coagulation. The laser irradiation time was 5 s. Vessel burst pressure measurements were used to determine seal strength. The 1470 nm laser wavelength demonstrated the capability of sealing a wide range of blood vessels from 1 to 6 mm diameter with burst strengths of 578±154 , 530±171 , and 426±174  mmHg for small, medium, and large vessel diameters, respectively. Lateral thermal coagulation zones (including the seal) measured 1.0±0.4  mm on vessels sealed at this wavelength. Other laser wavelengths (1550, 1850 to 1880, and 1908 nm) were also capable of sealing vessels, but were limited by lower vessel seal pressures, excessive charring, and/or limited power output preventing treatment of large vessels (<4  mm outer diameter).

Ocular prostheses are important determinants of their users’ aesthetic recovery and self-esteem. Because of use, ocular prostheses longevity is strongly affected by instability of the iris color due to polymerization. The goal of this study is to examine how the color of the artificial iris button is affected by different techniques of artificial wear and by the application of varnish following polymerization of the colorless acrylic resin that covers the colored paint. We produce 60 samples (n=10 ) according to the wear technique applied: conventional technique without varnish (PE); conventional technique with varnish (PEV); technique involving a prefabricated cap without varnish (CA); technique involving a prefabricated cap with varnish (CAV); technique involving inverted painting without varnish (PI); and technique involving inverted painting with varnish (PIV). Color readings using a spectrophotometer are taken before and after polymerization. We submitted the data obtained to analyses of variance and Tukey’s test (P<0.05 ). The color test shows significant changes after polymerization in all groups. The PE and PI techniques have clinically acceptable values of ΔE , independent of whether we apply varnish to protect the paint. The PI technique produces the least color change, whereas the PE and CA techniques significantly improve color stability.

Optical properties of cornea and sclera of the eye and their alterations under the effect of 1.56-μm laser radiation are studied. The laser settings corresponded to the laser treatment regimens used (1) to correct the shape of the cornea and change the refraction of the eye and (2) to improve the hydraulic permeability of the sclera in glaucoma cases. A fiber-optical system to investigate the dynamics of the reflected and transmitted scattered laser radiation and a setup with a double integrating sphere to determine the optical properties of the ocular tissues on the basis of the Monte-Carlo simulation of the propagation of light was used. When the radiation characteristics corresponded to the treatment regimens for correcting the shape of the cornea, no noticeable changes were detected in its optical properties. When irradiating the sclera in conditions corresponding to the treatment regimens for improving its hydraulic permeability, the optical characteristics of the tissue showed definite changes. The results obtained as to the dynamics of the optical signals during the course of laser irradiation of the cornea and sclera create prerequisites for designing test systems to be used with novel medical laser techniques for correcting visual abnormalities.

This article [J. Biomed. Opt.. 18, , 056010 (2013)] was originally published online on 9 May 2013 with an error in the author list: the fourth author’s name was misspelled as “Collard Dutilleul.” The correct spelling is “Collart Dutilleul,” as it appears above. This article was corrected online on 29th May 2013. It appears correctly in print.