Three-dimensional imaging of thick tissue constructs is one of the main challenges in the field of tissue engineering and regenerative medicine. Optical methods are the most promising as they offer noninvasive, fast, and inexpensive solutions. Herein, we report the use of mesoscopic fluorescence molecular tomography (MFMT) to image function and structure of thick bioprinted tissue hosted in a 3-mm-thick bioreactor. Collagen-based tissue assembled in this study contains two vascular channels formed by green fluorescent protein- and mCherry-expressing cells. Transfected live cell imaging enables us to image function, whereas Flash Red fluorescent bead perfusion into the vascular channel allows us to image structure. The MFMT optical reconstructions are benchmarked with classical microscopy techniques. MFMT and wide-field fluorescence microscopy data match within 92% in area and 84% in location, validating the accuracy of MFMT reconstructions. Our results demonstrate that MFMT is a well-suited imaging modality for fast, longitudinal, functional imaging of thick, and turbid tissue engineering constructs.

We have developed a novel miniature integrated optical coherence tomography (OCT)-intravascular ultrasound (IVUS) probe, with a 1.5-mm-long rigid part and 0.9-mm outer diameter, for real-time intracoronary imaging of atherosclerotic plaques and guiding of interventional procedures. By placing the OCT ball lens and IVUS transducer back-to-back at the same axial position, this probe can provide automatically coregistered, coaxial OCT-IVUS imaging. To demonstrate its real-time capability, three-dimensional OCT-IVUS imaging of a pig’s coronary artery displaying in polar coordinates, as well as images of three major types of atherosclerotic plaques in human cadaver coronary segments, were obtained using this probe and our upgraded system. Histology validation is also presented.

Iterative polynomial fitting along image rows and columns has recently been used to remove curvature bias in multispectral image sets of the human forearm and phantoms. However, this method is only applicable if foreground and background features satisfy strong separation conditions. In this method, we verify that the iterative polynomial approach converges toward bivariate polynomial fitting, and, hence, the resulting fit corresponds to low-pass filtering the image. In contrast to the iterative fitting, the bivariate polynomial fit can be performed on images with missing or excluded parts. Indeed, our observation enables us to modify the scheme and significantly weaken the required assumptions on foreground/background separation allowing a wider range of application.

We present a single-channel detection-based polarization-sensitive full-field optical coherence tomography (PS-FF-OCT) for simultaneous acquisition of high-resolution OCT and linear retardance images. A linearly polarized sub-10-fs laser was used as a broadband light source for the OCT system. A bi-stable polarization switching device, composed of a ferroelectric liquid crystal cell and an analyzer, was employed to get the horizontal and the vertical polarization components of a full-field interference signal. The time-switched two perpendicular interference signals were sequentially recorded by a single charge-coupled device camera, then processed to extract en-face functional images of a biological sample. The rat tail tendon was imaged ex vivo to confirm the imaging feasibility of the proposed system, which showed axial and transverse resolutions of 2 and 1.3 μm, respectively.

We present an improved optodynamic (OD) method which enables measurement of the distance between the OD source on the ablated surface and a piezoelectric sensor above it, with a relative error of about 1%. The method is based on the point explosion model and allows determination of the distance to the OD source and the released energy for each detected OD signal. We estimate the distance and released energy on the basis of two measured OD signal characteristics: the time of flight and the duration of the compressive phase. We show that the finite aperture of the sensor needs to be taken into account to improve measurement accuracy. We present experimental validation of the method using an Er:YAG laser and water as a tissue phantom. We observe an excellent agreement between the measured and theoretical OD signals and between the measured and estimated distances. The method opens the way to practicable implementations of on-line OD monitoring of laser ablation in surgery and medicine.

Spectral imaging is a technology that integrates conventional imaging and spectroscopy to get both spatial and spectral information from an object. Although this technology was originally developed for remote sensing, it has been extended to the biomedical engineering field as a powerful analytical tool for biological and biomedical research. This review introduces the basics of spectral imaging, imaging methods, current equipment, and recent advances in biomedical applications. The performance and analytical capabilities of spectral imaging systems for biological and biomedical imaging are discussed. In particular, the current achievements and limitations of this technology in biomedical engineering are presented. The benefits and development trends of biomedical spectral imaging are highlighted to provide the reader with an insight into the current technological advances and its potential for biomedical research.

Professor Roger Y. Tsien is a renowned world leader in chemical biology and biochemistry whose pioneering work in fluorescence sensing and imaging has revolutionized virtually all of experimental biology. He is an insightful scientist, and a cordial colleague and mentor; always invigorating with his ideas and energy, and inspiring with his vision. His work has profoundly influenced our abilities to study biochemical and molecular processes and events in live cells and animals. His early work on calcium imaging has revealed spatiotemporal characteristics of calcium signaling in various physiological processes and resulted in a widely disseminated use of his methods. But it is his seminal contributions in elucidating the biochemical mechanism of green fluorescent protein (GFP) formation that brought him major acclaim. Over a short period of time, Prof. Tsien further invented a broad spectrum of fluorescent proteins with distinctive colors and characteristics, a technology also adopted in all corners of the biology and biotechnology world. All these contributions have not only provided powerful tools for molecular and cellular imaging, but also advanced our in-depth and systematic understanding of a variety of cellular processes. To date, Prof. Tsien continues to lead and contribute to the emerging fields of molecular and cellular imaging as well as biomedical optics. Because of Prof. Tsien’s seminal scientific achievements, he was awarded the 2008 Nobel Prize in Chemistry for his role in “the green fluorescent protein: discovery, expression and development” and the Wolf Prize in medicine “for his seminal contribution to the design and biological application of novel fluorescent and photolabile molecules to analyze and perturb cell signal transduction.” Prof. Tsien is a member of the National Academy of Sciences and the Institute of Medicine, a Howard Hughes Medical Institute (HHMI) investigator, and a professor at University of California, San Diego.

The visual identification and demarcation of tumors and tumor margins remains challenging due to the low optical contrast of cancer cells over surrounding tissues. Fluorescence molecular imaging was recently considered clinically for improving cancer detection during open surgery. We present herein a next step in the development of fluorescence molecular guidance by describing a novel video-rate imaging laparoscope capable of concurrently recording color and near-infrared fluorescence images and video. Video-rate operation is based on graphics processing unit-based image processing. We examine the optical characteristics of the system developed and provide performance metrics related to intra-operative endoscopic guidance, showcased on phantoms and endoscopic color and fluorescence molecular imaging of tumors in a mouse model of the human disease.

We have developed a near-infrared (NIR) fluorescence goggle system based on the complementary metal–oxide–semiconductor active pixel sensor imaging and see-through display technologies. The fluorescence goggle system is a compact wearable intraoperative fluorescence imaging and display system that can guide surgery in real time. The goggle is capable of detecting fluorescence of indocyanine green solution in the picomolar range. Aided by NIR quantum dots, we successfully used the fluorescence goggle to guide sentinel lymph node mapping in a rat model. We further demonstrated the feasibility of using the fluorescence goggle in guiding surgical resection of breast cancer metastases in the liver in conjunction with NIR fluorescent probes. These results illustrate the diverse potential use of the goggle system in surgical procedures.

Antibody fragments including diabodies have more desirable pharmacokinetic characteristics than whole antibodies. An activatable optical imaging probe based on a cys-diabody targeting prostate-specific membrane antigen conjugated with the near-infrared fluorophore, indocyanine green (ICG), was designed such that it can only be activated when bound to the tumor, leading to high signal-to-background ratios. We employed short polyethylene glycol (PEG) linkers between the ICG and the reactive functional group (Sulfo-OSu group), resulting in covalent conjugation of ICG to the cys-diabody, which led to lower dissociation of ICG from cys-diabody early after injection, reducing hepatic uptake. However, unexpectedly, high and long-term fluorescence was observed in the kidneys, liver, and blood pool more than 1 h after injection of the cys-diabody PEG-ICG conjugate. A biodistribution study using I 125 -labeled cys-diabody-ICG showed immediate uptake in the kidneys followed by a rapid decrease, while gastric activity increased due to released radioiodine during rapid cys-diabody-ICG catabolism in the kidneys. To avoid this catabolic pathway, it would be preferable to use antibody fragments large enough not to be filtered through glomerulus or to conjugate the fragments with fluorescent dyes that are readily excreted into urine when cleaved from the cys-diabody to achieve high tumor-specific detection.

Fluorescence gene reporters have recently become available for excitation at far-red wavelengths, enabling opportunities for small animal in vivo gene reporter fluorescence tomography (GRFT). We employed multiple projections of the far-red fluorescence gene reporters IFP1.4 and iRFP, excited by a point source in transillumination geometry in order to reconstruct the location of orthotopically implanted human prostate cancer (PC3), which stably expresses the reporter. Reconstruction was performed using a linear radiative-transfer-based regularization-free tomographic method. Positron emission tomography (PET) imaging of a radiolabeled antibody-based agent that targeted epithelial cell adhesion molecule overexpressed on PC3 cells was used to confirm in vivo GRFT results. Validation of GRFT results was also conducted from ex vivo fluorescence imaging of resected prostate tumor. In addition, in mice with large primary prostate tumors, a combination of GRFT and PET showed that the radiolabeled antibody did not penetrate the tumor, consistent with known tumor transport limitations of large (∼150  kDa ) molecules. These results represent the first tomography of a living animal using far-red gene reporters.

The liver is important in the biotransformation of various drugs, where hepatic transporters facilitate uptake and excretion. Ischemia-reperfusion (I/R) injury is a common occurrence in liver surgery, and the developing oxidative stress can lead to graft failure. We used intravital multiphoton tomography, with fluorescence lifetime imaging, to characterize metabolic damage associated with hepatic I/R injury and to model the distribution of fluorescein as a measure of liver function. In addition to measuring a significant increase in serum alanine transaminase levels, characteristic of hepatic I/R injury, a decrease in the averaged weighted lifetime of reduced nicotinamide adenine dinucleotide phosphate was observed, which can be attributed to a changed metabolic redox state of the hepatocytes. I/R injury was associated with delayed uptake and excretion of fluorescein and elevated area-under-the-curve within the hepatocytes compared to sham (i.e., untreated control) as visualized and modeled using images recorded by intravital multiphoton tomography. High-performance liquid chromatography analysis showed no differences in plasma or bile concentrations of fluorescein. Finally, altered fluorescein distribution was associated with acute changes in the expression of liver transport proteins. In summary, multiphoton intravital imaging is an effective approach to measure liver function and is more sensitive in contrasting the impact of I/R injury than measuring plasma and bile concentrations of fluorescein.

Focused ultrasound (FUS) is a recently discovered noninvasive technique for local and temporal enhancement of vascular permeability, which facilitates drug delivery from the vessels into the surrounding tissue. However, exposure to FUS at a high intensity may cause permanent damage. To investigate the effects of the FUS treatment on blood vessels, we propose to use fluorescein angiography (FA) and optical coherence tomography (OCT) for real-time observation of the diffusion of fluorescence dye from blood vessels and to evaluate the morphological changes of the vessels in vivo. With time-resolved FA imaging, the relationship between the exposed power and the improved permeability of the vessels can be assessed according to the enhancement of the fluorescent intensity due to the dye leakage. Furthermore, the variation of the time-resolved fluorescent intensities can be used to identify the occurrence of dye leakage. In contrast, OCT can be implemented for the reconstruction of tissue microstructures. To quantitatively evaluate the morphological changes of the vessels after the FUS exposure with OCT, a new algorithm was proposed to estimate the vessel area based on the comparison of backscattering properties resulting from the tissue and vascular structures. Results showed that the vessel area increased as the exposed power increased, and the area became significantly larger at a higher FUS exposure power of 10 W. In conclusion, integrated FA and OCT observation can be potentially effective for monitoring the outcome and investigating the effects of FUS treatment.

Intervertebral disc degeneration (IDD) is closely associated with low back pain. Typically nonsurgical treatment of IDD is the most effective when detected early. As such, establishing reliable imaging methods for the early diagnosis of disc degeneration is critical. The cellular and tissue localization of a facile functional fluorescent probe, HYK52, that labels disc annulus fibrosus is reported. HYK52 was synthesized with high yield and purity via a two-step chemical reaction. Rabbit disc cell studies and ex vivo tissue staining images indicated intracellular localization and intervertebral disc (IVD) tissue binding of HYK52 with negligible cytotoxicity. Moreover, HYK52 is purposefully designed with a functional terminal carboxyl group to allow for coupling with various signaling molecules for multimodal imaging applications. These results suggest that this IVD-targeted probe may have great potential in early diagnosis of IDD.

The importance of guanine-quadruplex (G4) is not only in protecting the ends of chromosomes for human telomeres but also in regulating gene expression for several gene promoters. However, the existence of G4 structures in living cells is still in debate. A fluorescent probe, 3,6-bis(1-methyl-2-vinylpyridinium) carbazole diiodide (o-BMVC), for differentiating G4 structures from duplexes is characterized. o-BMVC has a large contrast in fluorescence decay time, binding affinity, and fluorescent intensity between G4 structures and duplexes, which makes it a good candidate for probing G4 DNA structures. The fluorescence decay time of o-BMVC upon interaction with G4 structures of telomeric G-rich sequences is ∼2.8  ns and that of interaction with the duplex structure of a calf thymus is ∼1.2  ns . By analyzing its fluorescence decay time and histogram, we were able to detect one G4 out of 1000 duplexes in vitro. Furthermore, by using fluorescence lifetime imaging microscopy, we demonstrated an innovative methodology for visualizing the localization of G4 structures as well as mapping the localization of different G4 structures in living cells.

An optical lensless configuration for a remote noncontact measuring of mechanical contractions of a vast number of cardiac myocytes is proposed. All the myocytes were taken from rats, and the measurements were done in an in vitro mode. The optical method is based on temporal analysis of secondary reflected speckle patterns generated in lensless microscope configuration. The processing involves analyzing the movement and the change in the statistics of the secondary speckle patterns that are created on top of the cell culture when it is illuminated by a spot of laser beam. The main advantage of the proposed system is the ability to measure many cells simultaneously (∼1000 cells) and to extract the statistical data of their movement at once. The presented experimental results also include investigation of the effect of isoproteranol on cell contraction process.

A new contrast agent, LipImage™ 815, has been designed and compared to previously described indocyanine green (ICG)-loaded lipid nanoparticles (ICG-lipidots®). Both contrast agents display similar size (50-nm diameter), zeta potential, high IC50 in cellular studies, near-infrared absorption and emission wavelengths in the “imaging window,” long-term shelf colloidal and optical stabilities with high brightness (>10 6   L mol −1  cm −1 ) in ready-to-use storage conditions in aqueous buffer (4°C in dark), therefore being promising fluorescence contrast agents for in vivo imaging. However, while ICG-lipidots® display a relatively short plasma lifetime, LipImage™ 815 circulates in blood for longer times, allowing the efficient uptake of fluorescence signal in human prostate cancer cells implanted in mice. Prolonged tumor labeling is observed for more than 21 days.

Design and development of a new formulation as a unique assembly of distinct fluorescent reporters with nonoverlapping fluorescence spectra and a F 19 magnetic resonance imaging agent into colloidally and optically stable triphasic nanoemulsion are reported. Specifically, a cyanine dye-perfluorocarbon (PFC) conjugate was introduced into the PFC phase of the nanoemulsion and a near-infrared dye was introduced into the hydrocarbon (HC) layer. To the best of our knowledge, this is the first report of a triphasic nanoemulsion system where each oil phase, HC, and PFC are fluorescently labeled and formulated into an optically and colloidally stable nanosystem. Having, each oil phase separately labeled by a fluorescent dye allows for improved correlation between in vivo imaging and histological data. Further, dual fluorescent labeling can improve intracellular tracking of the nanodroplets and help assess the fate of the nanoemulsion in biologically relevant media. The nanoemulsions were produced by high shear processing (microfluidization) and stabilized with biocompatible nonionic surfactants resulting in mono-modal size distribution with average droplet size less than 200 nm. Nanoemulsions demonstrate excellent colloidal stability and only moderate changes in the fluorescence signal for both dyes. Confocal fluorescence microscopy of macrophages exposed to nanoemulsions shows the presence of both fluorescence agents in the cytoplasm.

A spectral correlation algorithm for the analysis of hyperspectral fluorescence images is proposed by Ellingsen et al. [J. Biomed. Opt. 18, 020501 (2013)]. Here, it is applied to the analysis of double-stained Aβ amyloid plaques being related to the Alzheimer’s disease (AD). Sections of APP/PS1 AD mice model brains are double stained with luminescent-conjugated oligothiophenes, known to bind to amyloid protein deposits. Hyperspectral fluorescence images of the brain sections are recorded and by applying the correlation algorithm the spectral inhomogeneity of the double-stained samples is mapped in terms of radial distribution and spectral content. To further investigate the progression of Aβ amyloid plaque formation, 19 AD mice of different ages up to 23 months are characterized, enabling a statistical analysis of the plaque heterogeneity. In accordance with recent findings by Nyström et al. [ACS Chem. Biol. 8, 1128–1133 (2013)], the spectral distribution within Aβ plaques is found to vary with age throughout the lifespan of the mouse. With the new correlation algorithm, it is possible to quantify the spectral abundance of the two stains depending on the relative distance from the plaque center and mouse age. Thus, we demonstrate the use of the correlation analysis approach in double-staining experiments and how it is possible to relate these to structural/spectral changes in biological samples.

We demonstrate the use of an enzyme-activatable fluorogenic probe, Neutrophil Elastase 680 FAST (NE680), for in vivo imaging of neutrophil elastase (NE) activity in tumors subjected to photodynamic therapy (PDT). NE protease activity was assayed in SCC VII and EMT6 tumors established in C3H and BALB/c mice, respectively. Four nanomoles of NE680 was injected intravenously immediately following PDT irradiation. 5 h following administration of NE680, whole-mouse fluorescence imaging was performed. At this time point, levels of NE680 fluorescence were at least threefold greater in irradiated versus unirradiated SCC VII and EMT6 tumors sensitized with Photofrin. To compare possible photosensitizer-specific differences in therapy-induced elastase activity, EMT6 tumors were also subjected to 2-(1-hexyloxyethyl)-2-devinyl pyropheophorbide-a (HPPH)-PDT. NE levels measured in HPPH-PDT-treated tumors were twofold higher than in unirradiated controls. Ex vivo labeling of host cells using fluorophore-conjugated antibodies and confocal imaging were used to visualize Gr1 + cells in Photofrin-PDT-treated EMT6 tumors. These data were compared with recently reported analysis of Gr1 + cell accumulation in EMT6 tumors subjected to HPPH-PDT. The population density of infiltrating Gr1 + cells in treated versus unirradiated drug-only control tumors suggests that the differential in NE680 fold enhancement observed in Photofrin versus HPPH treatment may be attributed to the significantly increased inflammatory response induced by Photofrin-PDT. The in vivo imaging of NE680, which is a fluorescent reporter of NE extracellular release caused by neutrophil activation, demonstrates that PDT results in increased NE levels in treated tumors, and the accumulation of the cleaved probe tracks qualitatively with the intratumor Gr1 + cell population.

The optical properties of a receptor-targeted probe designed for dual-modality mapping of the sentinel lymph node (SLN) was optimized. Specific fluorescence brightness was used as the design criterion, which was defined as the fluorescence brightness per mole of the contrast agent. Adjusting the molar ratio of the coupling reactants, IRDye 800CW-NHS-ester and tilmanocept, enabled us to control the number of fluorescent molecules attached to each tilmanocept, which was quantified by H 1 nuclear magnetic resonance spectroscopy. Quantum yields and molar absorptivities were measured for unconjugated IRDye 800CW and IRDye 800CW-tilmanocept (800CW-tilmanocept) preparations at 0.7, 1.5, 2.3, 2.9, and 3.8 dyes per tilmanocept. Specific fluorescence brightness was calculated by multiplication of the quantum yield by the molar absorptivity and the number of dyes per tilmanocept. It predicted that the preparation with 2.3 dyes per tilmanocept would exhibit the brightest signal, which was confirmed by fluorescence intensity measurements using three optical imaging systems. When radiolabeled with Ga 68 and injected into the footpads of mice, the probe identified SLNs by both fluorescence and positron emission tomography (PET) while maintaining high percent extraction by the SLN. These studies demonstrated the feasibility of 800CW-tilmanocept for multimodal SLN mapping via fluorescence and PET–computed tomography imaging.

Förster resonance energy transfer (FRET) is a distance-dependent process that transfers excited state energy from a donor molecule to an acceptor molecule without the emission of a photon. The FRET rate is determined by the proximity between the donor and the acceptor molecules; it becomes significant only when the proximity is within several nanometers. Therefore, FRET has been applied to visualize interactions and conformational changes of biomolecules, such as proteins, lipids, and nucleic acids that cannot be resolved by optical microscopy. Here, we report photoacoustic tomography of FRET efficiency at a 1-cm depth in chicken breast tissue, whereas conventional high-resolution fluorescence imaging is limited to <0.1  cm . Photoacoustic tomography is expected to facilitate the examination of FRET phenomena in living organisms.

An ultra-sensitive hyphenated technique, high-performance liquid chromatography-laser-induced fluorescence detection protein profiling of saliva, is evaluated for early detection and diagnosis of oral premalignancy and malignancy. Calibration sets of protein profiles of unstimulated whole saliva are collected from clinically/pathologically normal, premalignant, and malignant subjects and used as standards. Three parameters—scores of factors, sum of squared residuals, and Mahalanobis distance—derived from principal component analysis of protein profiles of the standard calibration sets, and blind samples are used for “match/no-match” diagnosis of the blind samples. Analyses of the results show that the method is capable of differentiating normal, premalignant, and malignant conditions with the sensitivity and specificity of 79% and 78%, respectively. The technique provides a fast, highly objective (free from personal judgment and statistically defined), and noninvasive diagnostic method for screening and early detection of oral cancer.

To minimize the problem with scattering in deep tissues while increasing the penetration depth, we explored the feasibility of imaging in the relatively unexplored extended near infrared (exNIR) spectral region at 900 to 1400 nm with endogenous chromophores. This region, also known as the second NIR window, is weakly dominated by absorption from water and lipids and is free from other endogenous chromophores with virtually no autofluorescence. To demonstrate the applicability of the exNIR for bioimaging, we analyzed the optical properties of individual components and biological tissues using an InGaAs spectrophotometer and a multispectral InGaAs scanning imager featuring transmission geometry. Based on the differences in spectral properties of tissues, we utilized ratiometric approaches to extract spectral characteristics from the acquired three-dimensional “datacube”. The obtained images of an exNIR transmission through a mouse head revealed sufficient details consistent with anatomical structures.

Assays for blood levels of prostate-specific antigen (PSA), performed in prostate cancer detection, measure mostly inactive/complexed PSA and do not provide information regarding enzymatically active PSA, which is biologically more relevant. Thus, we designed and synthesized an enzymatically cleavable peptide sequence labeled with near-infrared (NIR) fluorophores (ex/em 740/770  nm ) and coupled it to a pharmacokinetic modifier designed to improve its plasma kinetics. In its native state, the agent, PSA750 FAST™ (PSA750), is optically quenched (>95% ) and only becomes fluorescent upon cleavage by active PSA, yielding a significant increase in signal. This activation is highly selective for PSA relative to a large panel of disease-relevant enzymes. Active PSA was detected in tumor frozen sections using PSA750 and this activity was abolished in the presence of the inhibitor, alpha-1 anti-chymotrypsin. In vivo imaging of tumor-bearing mice using fluorescence molecular tomography demonstrated a significantly higher fluorescent signal in PSA + LNCaP tumors as compared to PSA − prostate cancer 3 tumors (13.0±3.7 versus 2.8±0.8  pmol , p=0.023 ). Ex vivo imaging of tumor sections confirms PSA750-derived NIR signal localization in nonvascular tissue. This is the first report that demonstrates the feasibility and effectiveness of noninvasive, real time, fluorescence molecular imaging of PSA enzymatic activity in prostate cancer.

Optical clearing agents can improve tissue optical transmittance by reducing the diffuse reflection. The reflection on in vivo human skin before and after applying anhydrous glycerol and 30 to 50% liquid paraffin glycerol mixed solution are investigated in this paper. From their visible and near-infrared reflection spectroscopy, all of their diffuse reflections are reduced after applying the agents. It is found that the three mixed solutions show stronger effect than that of anhydrous glycerol. These results further prove liquid paraffin can enhance the percutaneous penetration of glycerol and take synergistically optical clearing effect with glycerol over visible and near-infrared wave bands.

Technologies currently available for the monitoring of electrical stimulation (ES) in promoting blood circulation and tissue oxygenation are limited. This study integrated a muscle stimulator with a diffuse correlation spectroscopy (DCS) flow-oximeter to noninvasively quantify muscle blood flow and oxygenation responses during ES. Ten healthy subjects were tested using the integrated system. The muscle stimulator delivered biphasic electrical current to right leg quadriceps muscle, and a custom-made DCS flow-oximeter was used for simultaneous measurements of muscle blood flow and oxygenation in both legs. To minimize motion artifact of muscle fibers during ES, a novel gating algorithm was developed for data acquisition at the time when the muscle was relaxed. ES at 2, 10, and 50 Hz were applied for 20 min on each subject in three days sequentially. Results demonstrate that the 20-min ES at all frequencies promoted muscle blood flow significantly. However, only the ES at 10 Hz resulted in significant and persistent increases in oxy-hemoglobin concentration during and post ES. This pilot study supports the application of the integrated system to quantify tissue hemodynamic improvements for the optimization of ES treatment in patients suffering from diseases caused by poor blood circulation and low tissue oxygenation (e.g., pressure ulcer).

The depth distributions of refractive index (RI) in normal and early degenerated articular cartilage (AC) were measured and correlated to the pathological statuses. Utilizing a confocal microscope, the depth distribution of RI was determined with an interval of 50 μm from the articular surface and approximated as a smooth curve. The fluctuation of RI was quantified with a specially defined coefficient and the proteoglycan (PG) loss was quantified with histological images. The overall RI of the AC collected from the femoral condyles of mature sheep was 1.4444. Significant differences existed in the refractive indices of different cartilage samples or even in the different depths of the same sample. The RI of normal cartilage distributed as a regular ramp or arch curve with depth and peaked in the middle–deep zone. The cartilage samples detected with high PG losses were coupled with severe fluctuations of RI. The fluctuating level was increased with pathological progress. The coefficient of RI fluctuation can distinguish the normal cartilage from the initially degenerated one, which potentially provides an indicator to detect the pathological development of AC.

Continuous-wave near-infrared spectroscopy and near-infrared imaging enable the measurement of relative concentration changes in oxy- and deoxyhemoglobin and thus hemodynamics and oxygenation. The accuracy of determined changes depends mainly on the modeling of the light transport through the probed tissue. Due to the highly scattering nature of tissue, the light path is longer than the source–detector separation (d ). This is incorporated in modeling by multiplying d by a differential pathlength factor (DPF) which depends on several factors such as wavelength, age of the subject, and type of tissue. In the present work, we derive a general DPF equation for the frontal human head, incorporating dependency on wavelength and age, based on published data. We validated the equation using different data sets of experimentally determined DPFs from six independent studies.

The effect of different beverages on acrylic resin denture teeth color degradation is evaluated. Ten acrylic resin denture teeth brands were evaluated: Art Plus (AP), Biolux (BX), Biotone IPN (BI), Magister (MG), Mondial 6 (MD), Premium 6 (PR), SR Vivodent PE (SR), Trilux (TR), Trubyte Biotone (TB), and Vipi Dent Plus (VP). Teeth were immersed in staining solutions (coffee, cola, and orange juice) or artificial saliva (control) (n=6 ) for 1, 7, 15, or 30 days. Specimen colors were evaluated spectrophotometrically based on the Commission Internationale d’Eclairage L*a*b* system. Color differences (ΔE ) were calculated between the baseline and post-staining results. Data were evaluated by analysis of variance and Tukey test (α=0.05 ). BI (1.82±0.95 ) and TR (1.78±0.72 ) teeth exhibited the greatest ΔE values, while BX (0.88±0.43 ) and MD (1.09±0.44 ) teeth were the lowest, regardless of solution and measurement period, and were different from BI and TR teeth (P<0.05 ). Cola and coffee promoted higher denture teeth color alterations than orange juice and saliva (P<0.05 ). Saliva generated the lowest denture teeth color alterations. Greater immersion times caused higher denture teeth color changes. The lifespan of removable dentures and the aesthetic satisfaction of several edentulous patients may be increased with the use of stain-resistant artificial denture teeth.

We present a two-dimensional electromagnetic analysis of light propagation through the human eye to examine the eye’s optical properties. The electromagnetic approach has intriguing advantages over the conventional and frequently implemented ray optics analysis. The chromatic, spherical, and coma aberrations and the intensity of the focused light at the retina are computed in this work via full-wave analysis. We also investigate the effects of the cornea’s and lens’s curved structures on the focusing mechanism. The focal length and chromatic and spherical aberrations are observed to change owing to age-related refractive index variation in the lens. In addition, the effects of the lens and curvatures of the human eye on focusing are analyzed. Consequently, for both young and old human eye lenses, the differences due to the aberration variations, curvature surfaces, and gradient index are explored by the wave approach. The intensity distributions on the retina for both on- and off-axis illumination are calculated. A strong correlation between the locations of the nerve fibers and the intensity distribution is confirmed. On the basis of the findings, we can conclude that visual impairment due to deterioration of the human eye structure is more dramatic than that due to aging.

Noninvasive blood analysis devices that can measure levels of small constituents of blood are of interest in the medical community. An important step in creating these devices is to understand the interaction of photons with human tissue in increasingly greater physiological detail. Models based on layered biological materials give excellent results for many applications but may not be as accurate as needed when those materials are finely intertwined to the point of resembling a homogeneous mixture. To explore the ramifications of treating materials as layers versus a mixture, we have modeled, using a Monte Carlo technique, the interaction of photons through epidermis, blood, and water arranged both in layers and in a homogeneous blend. We confirm the expected linear relation between photon attenuation and material volumetric percentage in two-layer models. However, when the materials are homogeneously mixed together and volumetric percentage is replaced with interaction volume percentage, this relationship becomes nonlinear. These nonlinearities become significant when the values of the interaction coefficient, μ t , differ by an order of magnitude or more.

The diagnosis and medical treatment of cerebral ischemia are becoming more important due to the increase in the prevalence of cerebrovascular disease. However, conventional methods of evaluating cerebral perfusion have several drawbacks: they are invasive, require physical restraint, and the equipment is not portable, which makes repeated measurements at the bedside difficult. An alternative method is developed using near-infrared spectroscopy (NIRS). NIRS signals are measured at 44 positions (22 on each side) on the fronto-temporal areas in 20 patients with cerebral ischemia. In order to extract the pulse-wave component, the raw total hemoglobin data recorded from each position are band-pass filtered (0.8 to 2.0 Hz) and subjected to a fast Fourier transform to obtain the power spectrum of the pulse wave. The ischemic region is determined by single-photon emission computed tomography. The pulse-wave power in the ischemic region is compared with that in the symmetrical region on the contralateral side. In 17 cases (85%), the pulse-wave power on the ischemic side is significantly lower than that on the contralateral side, which indicates that the transmission of the pulse wave is attenuated in the region with reduced blood flow. Pulse-wave power might be useful as a noninvasive marker of cerebral ischemia.

We present the application of two-photon fluorescence (TPF) imaging to monitor intracellular hydrogen peroxide (H 2 O 2 ) production in brain cells. For selective imaging of H 2 O 2 over other reactive oxygen species, we employed small-molecule fluorescent probes that utilize a chemoselective boronate deprotection mechanism. Peroxyfluor-6 acetoxymethyl ester detects global cellular H 2 O 2 and mitochondria peroxy yellow 1 detects mitochondrial H 2 O 2 . Two-photon absorption cross sections for these H 2 O 2 probes are measured with a mode-locked Ti:sapphire laser in the wavelength range of 720 to 1040 nm. TPF imaging is demonstrated in the HT22 cell line to monitor both cytoplasmic H 2 O 2 and localized H 2 O 2 production in mitochondria. Endogenous cytoplasmic H 2 O 2 production is detected with TPF imaging in rat astrocytes modified with d-amino acid oxidase. The TPF H 2 O 2 imaging demonstrated that these chemoselective probes are powerful tools for the detection of intracellular H 2 O 2 .

As in vivo flow behavior can be pulsatile, intermittent, and/or otherwise changeable with time, the ability to provide clinicians with a means of real-time visualization and functional assessment of structures is of particular importance. The discernment of pulsatile flow behavior using a dual-beam spectral domain optical coherence tomography system (db-SdOCT) by quasi-simultaneous measurement by two planes of illumination is demonstrated. By cross-correlation analysis, it is possible to compute velocity metrics pertaining to flowing particle motion, without a priori angular knowledge. This is the first application of cross-correlation-based dynamic assessment for the extraction of pulsatile behavior in an in vitro environment using an optimized db-SdOCT system. The experimental results outlined have shown the db-SdOCT system and its associated algorithms to be successful in the discernment of intermittent pulsatile flow behavior in in vitro models, concurrent to yielding velocity values in good agreement with that of the applied flow rate.

We present an innovative method, photoacoustic recovery after photothermal bleaching (PRAP), for studying particle dynamics at micron scale via photoacoustic imaging. As an intuitive way to visualize and quantify dynamic processes, PRAP is demonstrated first in a simple phantom study and then in a more complex measurement involving live cells. Compared with the conventional fluorescence-based approach, PRAP provides high signal-to-noise ratio (SNR) imaging with minimal bleaching-induced artifacts during the recovery stage, ideal for monitoring the diffusive and kinetic processes inside a cell.

Optoacoustic (photoacoustic) mesoscopy aims at high-resolution optical imaging of anatomical, functional, and cellular parameters at depths that go well beyond those of optical-resolution optical or optoacoustic microscopy i.e., reaching several millimeters in depth. The approach utilizes tomography to achieve ultrasonic-diffraction resolution and operates at high-ultrasound frequencies (20 to 200 MHz) induced by few-nanosecond laser pulse excitation of tissues. We investigated here the performance of optoacoustic mesoscopy implemented at 24 MHz center frequency and its ability to resolve optical absorption contrast in the mouse kidney ex vivo. The developed system achieved better than 30 μm in-plane resolution and 110 μm elevation resolution over a cylindrical volume of 9-mm diameter and 9-mm height. This unprecedented combination of resolution and depth was achieved by implementing a translate-rotate detection geometry and by tomographic reconstruction. The approach yielded images of optically absorbing structures with a level of detail never-before visualized in an intact mouse kidney and allows insights into their unperturbed architecture. We discuss the ability to offer multispectral acquisitions and enable in vivo imaging.

The dependence of the transition between the ballistic and the diffusive regimes of turbid media on the experimental solid angle of the detection system is analyzed theoretically and experimentally. A simple model is developed which shows the significance of experimental conditions on the location of the ballistic–diffusive transition. It is demonstrated that decreasing the solid angle expands the ballistic regime; however, this benefit is bounded by the initial Gaussian beam diffraction. In addition, choosing the appropriate wavelength according to the model’s principles provides another means of expanding the ballistic regime. Consequently, by optimizing the experimental conditions, it should be possible to extract the ballistic image of a tissue with a thickness of 1 cm.

Autofluorescence (AF) imaging provides valuable information about the structural and chemical states of tissue that can be used for early cancer detection. Optical scattering and absorption of excitation and emission light by the epithelium can significantly affect observed tissue AF intensity. Determining the effect of epithelial attenuation on the AF intensity could lead to a more accurate interpretation of AF intensity. We propose to use optical coherence tomography coregistered with AF imaging to characterize the AF attenuation due to the epithelium. We present imaging results from three vital tissue models, each consisting of a three-dimensional tissue culture grown from one of three epithelial cell lines (HCT116, OVCAR8, and MCF7) and immobilized on a fluorescence substrate. The AF loss profiles in the tissue layer show two different regimes, each approximately linearly decreasing with thickness. For thin cell cultures (<300  μm ), the AF signal changes as AF(t)/AF(0)=1−1.3t (t is the thickness in millimeter). For thick cell cultures (>400  μm ), the AF loss profiles have different intercepts but similar slopes. The data presented here can be used to estimate AF loss due to a change in the epithelial layer thickness and potentially to reduce AF bronchoscopy false positives due to inflammation and non-neoplastic epithelial thickening.

Noninvasive or minimally invasive identification of sentinel lymph node (SLN) is essential to reduce the surgical effects of SLN biopsy. Photoacoustic (PA) imaging of SLN in animal models has shown its promise for clinical use in the future. Here, we present a Monte Carlo simulation for light transport in the SLN for various light delivery configurations with a clinical ultrasound probe. Our simulation assumes a realistic tissue layer model and also can handle the transmission/reflectance at SLN-tissue boundary due to the mismatch of refractive index. Various light incidence angles show that for deeply situated SLNs the maximum absorption of light in the SLN is for normal incidence. We also show that if a part of the diffused reflected photons is reflected back into the skin using a reflector, the absorption of light in the SLN can be increased significantly to enhance the PA signal.

This study aimed to evaluate the stress distribution through the photoelastic method in implant-retained palatal obturators prostheses. Two photoelastic models with bucco-sinusal communication were fabricated, one model without implants and another with two parallel implants and one tilted in the molar region. A conventional obturator prosthesis and five implant-retained obturators dentures with different attachment systems were fabricated: OR, three individualized O-rings; BC, bar clip; BOC, implants splinted by bars associated with two O-rings positioned at the center of the bar; OD, implants splinted by bars associated with two O-rings positioned in distal cantilever; and BOD, implants splinted by bars with clips associated with two O-rings positioned in distal cantilever. Each assembly (model/attachment system/prosthesis) was positioned in a circular polariscope and a load of 100 N was applied on each implant. The results were obtained by observing the photographic record of the tensions in the photoelastic models resulting from the application of load. It can be observed that a larger amount of stress fringes on BC system. It was concluded that the attachment system has a direct influence on the stress distribution of implant-retained obturator prostheses, with the three individualized O-rings exhibiting the lowest stress values, and tilted implants presented a biomechanical behavior similar to parallel implants.

We present a time-domain polarization-sensitive (PS) optical coherence tomography configuration operating at 830 nm, equipped with multichannel acousto-optic deflectors and single photodetectors. The system is used to simultaneously acquire interference information from multiple PS channels and to enable measurement and imaging of backscattered intensity to create both PS and polarization insensitive images. Our approach enables multiple channel imaging without need to divide the object signal. Here, we employ our system in order to demonstrate PS imaging of a thermally damaged muscle tissue.

Functional near infrared spectroscopy (fNIRS) is a noninvasive method to capture brain activities according to the measurements of changes in both oxyhemoglobin and deoxyhemoglobin concentrations. However, fNIRS recordings are the hemodynamic signals that come from the latent neural sources that are spatially and temporally mixed across the brain. The purpose of this work is to extract the temporal and frequency characteristics as well as the spatial activation patterns in the brains using independent component analysis (ICA). In this study, the filtered fNIRS recordings were processed and the time-frequency and spatiotemporal domain independent components (ICs) were identified by ICA. We found that multiple task-related components can be separated by ICA in time-frequency domain, and distinct spatial patterns of brain activity can be derived from ICs that are well correlated with the specific neural events, such as finger tapping tasks.

The era of molecular medicine has ushered in the development of microscopic methods that can report molecular processes in thick tissues with high spatial resolution. A commonality in deep-tissue microscopy is the use of near-infrared (NIR) lasers with single- or multiphoton excitations. However, the relationship between different NIR excitation microscopic techniques and the imaging depths in tissue has not been established. We compared such depth limits for three NIR excitation techniques: NIR single-photon confocal microscopy (NIR SPCM), NIR multiphoton excitation with visible detection (NIR/VIS MPM), and all-NIR multiphoton excitation with NIR detection (NIR/NIR MPM). Homologous cyanine dyes provided the fluorescence. Intact kidneys were harvested after administration of kidney-clearing cyanine dyes in mice. NIR SPCM and NIR/VIS MPM achieved similar maximum imaging depth of ∼100  μm . The NIR/NIR MPM enabled greater than fivefold imaging depth (>500  μm ) using the harvested kidneys. Although the NIR/NIR MPM used 1550-nm excitation where water absorption is relatively high, cell viability and histology studies demonstrate that the laser did not induce photothermal damage at the low laser powers used for the kidney imaging. This study provides guidance on the imaging depth capabilities of NIR excitation-based microscopic techniques and reveals the potential to multiplex information using these platforms.

Stiles–Crawford effect (SCE) is exclusively observed in cone photoreceptors, but why the SCE is absent in rod photoreceptors is still a mystery. In this study, we employed dynamic near infrared light imaging to monitor photoreceptor kinetics in freshly isolated frog and mouse retinas stimulated by oblique visible light flashes. It was observed that retinal rods could rapidly (onset: ∼10  ms for frog and 5 ms for mouse; time-to-peak: ∼200  ms for frog and 30 ms for mouse) shift toward the direction of the visible light, which might quickly compensate for the loss of luminous efficiency due to oblique illumination. In contrast, such directional movement was negligible in retinal cones. Moreover, transient rod phototropism could contribute to characteristic intrinsic optical signal (IOS). We anticipate that further study of the transient rod phototropism may not only provide insight into better understanding of the nature of vision but also promise an IOS biomarker for functional mapping of rod physiology at high resolution.

Peripheral nerve injury in vivo promotes a regenerative growth in vitro characterized by an improved neurite regrowth. Knowledge of the conditioning injury effects on both morphology and mechanical properties of live sensory neurons could be instrumental to understand the cellular and molecular mechanisms leading to this regenerative growth. In the present study, we use differential interference contrast microscopy, fluorescence microscopy, and atomic force microscopy (AFM) to show that conditioned axotomy, induced by sciatic nerve injury, does not increase somatic size of sensory neurons from adult mice lumbar dorsal root ganglia but promotes the appearance of longer and larger neurites and growth cones. AFM on live neurons is also employed to investigate changes in morphology and membrane mechanical properties of somas of conditioned neurons following sciatic nerve injury. Mechanical analysis of the soma allows distinguishing neurons having a regenerative growth from control ones, although they show similar shapes and sizes.

We propose a simple and optimized method for acquiring a wide velocity range of blood flow using Doppler optical microangiography. After characterizing the behavior of the scanner in the fast scan axis, a step-scanning protocol is developed by utilizing repeated A-scans at each step. Multiple velocity range images are obtained by the high-pass filtering and Doppler processing of complex signals between A-scans within each step with different time intervals. A phase variance mask is then employed to segment meaningful Doppler flow signals from noisy phase background. The technique is demonstrated by imaging in vivo mouse brain with skull left intact to provide bidirectional images of cerebral blood flow with high quality and wide velocity range.

Breast cancer management could be improved by developing real-time imaging tools to assess tissue architecture without extensive processing. We sought to determine whether confocal fluorescence microscopy (CFM) provides sufficient information to identify neoplasia in breast tissue. Breast tissue specimens were imaged following proflavine application. Regions of interest (ROIs) were selected in histologic slides and in the corresponding region on confocal images, and then divided into sets for training and validation. Readers reviewed images in the training set and evaluated images in the validation set for the presence of neoplasia. Accuracy was assessed using histologic diagnosis as the gold standard. Seventy tissue specimens from 31 patients were imaged; 235 ROIs were identified and diagnosed as neoplastic or non-neoplastic. A training set was assembled using 23 matched ROIs; 49 matched ROIs were assembled into a validation set. Neoplasia was identified in histologic images: 93% sensitivity, 97% specificity [area under the curve (AUC=0.987 )] and in confocal images: 93% sensitivity 93% specificity (AUC=0.957 ). CFM produced images of architectural features in breast tissue comparable with conventional histology, while requiring little processing. Potential applications include assessment of excised tissue margins and evaluation of tissue adequacy for bio-banking and genomic studies.

A cost-efficient plastic optical fiber (POF) system for unobtrusive monitoring of human vital signs is presented. The system is based on speckle interferometry. A laser diode is butt-coupled to the POF whose exit face projects speckle patterns onto a linear optical sensor array. Sequences of acquired speckle images are transformed into one-dimensional signals by using the phase-shifting method. The signals are analyzed by band-pass filtering and a Morlet-wavelet-based multiresolutional approach for the detection of cardiac and respiratory activities, respectively. The system is tested with 10 healthy nonhospitalized persons, lying supine on a mattress with the embedded POF. Experimental results are assessed statistically: precisions of 98.8%±1.5% and 97.9%±2.3% , sensitivities of 99.4%±0.6% and 95.3%±3% , and mean delays between interferometric detections and corresponding referential signals of 116.6±55.5 and 1299.2±437.3  ms for the heartbeat and respiration are obtained, respectively.

A versatile, continuous wave, optical parametric oscillator is used in combination with photoacoustic spectroscopy for long-term trace gas experiments of volatile compounds emitted by biological samples. The optical parametric oscillator-based spectrometer (wavelength near 3 μm, 8-MHz linewidth, output power ∼1  W ) is successfully tested for the detection of hydrogen cyanide (HCN) emission from clover leaves, and Pseudomonas bacteria; in addition, the presence of HCN in exhaled human breath is measured. For specific experiments, the spectrometer is operated continuously up to 10 days and has a detection limit of 0.4 parts-per-billion volume of HCN in air over 10 s, using the P8 rotational line in the ν 3 vibrational band of HCN at 3287.25  cm −1 . This results in an overall sensitivity of the system of 2.5×10 −9   cm −1  Hz −1/2 .

Simplified measurement of macular pigment optical density (MPOD) is important because of the ocular health benefits that are attributed to these retinal carotenoids. Here, we describe a novel instrument designed for this purpose, based on heterochromatic flicker photometry (HFP), which removes a number of difficulties that subjects often experience with traditional HFP. The instrument generates 1.5- and 15-deg diameter, centrally viewed stimuli that alternate between blue and green colors generated by light emitting diodes (LED). The 15 deg stimulus replaces the small, eccentrically viewed stimulus used in traditional HFP. Subjects adjust the blue LED intensity until flicker is eliminated in the case of the 1.5 deg stimulus and eliminated around the periphery in the case of the 15 deg stimulus. A microprocessor computes the subject’s MPOD, in addition to the lens OD, and uses the latter to correct the MPOD. Good repeatability was confirmed through test–retest measurements on 52 subjects. The overwhelming majority of them stated that they found the test easy. The importance of the lens correction on MPOD measurements was confirmed in a simulation study. The study showed that, without the correction, MPOD would show an apparent age-related decline in a population for whom there was no real age dependence.

Optical biopsy techniques offer a minimally invasive, real-time alternative to traditional biopsy and pathology during tumor resection surgery. Diffuse reflectance spectroscopy (DRS) is a commonly used technique in optical biopsy. Optical property recovery from spatially resolved DRS data allows quantification of the scattering and absorption properties of tissue. Monte Carlo simulation methods were used to evaluate a unique fiber-optic probe design for a DRS instrument to be used specifically for optical biopsy of the brain. The probe diameter was kept to a minimum to allow usage in small surgical cavities at least 1 cm in diameter. Simulations showed that the close proximity of fibers to the edge of the probe resulted in boundary effects due to reflection of photons from the surrounding air–tissue interface. A new algorithm for rapid optical property recovery was developed that accounts for this reflection and therefore overcomes these effects. The parameters of the algorithm were adjusted for use over the wide range of optical properties encountered in brain tissue, and its precision was evaluated by subjecting it to random noise. This algorithm can be adapted to work with any probe geometry to allow optical property recovery in small surgical cavities.

We have recently demonstrated a means for quantifying the absorption and scattering properties of biological tissue through multidiameter single-fiber reflectance (MDSFR) spectroscopy. These measurements can be used to correct single-fiber fluorescence (SFF) spectra for the influence of optical properties, enabling quantification of intrinsic fluorescence. In our previous work, we have used a series of pinholes to show that selective illumination and light collection using a coherent fiber bundle can simulate a single solid-core optical fiber with variable diameter for the purposes of MDSFR spectroscopy. Here, we describe the construction and validation of a clinical MDSFR/SFF spectroscopy system that avoids the limitations encountered with pinholes and free-space optics. During one measurement, the new system acquires reflectance spectra at the effective diameters of 200, 600, and 1000 μm, and a fluorescence spectrum at an effective diameter of 1000 μm. From these spectra, we measure the absolute absorption coefficient, μ a , reduced scattering coefficient, μ ′ s , phase function parameter, γ , and intrinsic fluorescence, Qμ f a,x , across the measured spectrum. We validate the system using Intralipid- and polystyrene sphere-based scattering phantoms, with and without the addition of the absorber Evans Blue. Finally, we demonstrate the combined MDSFR/SFF of phantoms with varying concentrations of Intralipid and fluorescein, wherein the scattering properties are measured by MDSFR and used to correct the SFF spectrum for accurate quantification of Qμ f a,x .

Laser-induced breakdown spectroscopy (LIBS) is applied to investigate the effect of diabetes mellitus (DM) on the elemental composition of fingernails. Measurements are carried out on 85 fingernail clippings including 51 diabetic and 34 control subjects. An auto-focus system has been designed and used in experiments to improve the repeatability of LIBS measurements. Classification of diabetic and nondiabetic subjects is examined using discriminant function analysis (DFA) method. This classification is based on 82 atomic, ionic, and molecular emission lines belonging to 13 elements as well as one molecule of fingernails. Emission lines that can be used as the best predictors are identified. The possibility of using this method for screening purposes is discussed based on the classification results. This preliminary work shows the ability of LIBS of fingernails in discrimination of DM patients and nondiabetic subjects using DFA method and its feasibility in screening purposes.

We demonstrate interstitial recovery of absorption and scattering coefficients using a custom optical probe and a Monte Carlo (MC)–based recovery algorithm. The probe consists of six side-firing spectroscopy fibers contained in a 1.1-mm outer diameter cladding, with each fiber having a different axial and angular position on the probe. Broadband white light is delivered by one of the fibers and is detected steady-state by the remaining fibers. These spatially and spectrally resolved data are analyzed using a MC-based fitting algorithm in order to extract the local optical properties. The technique was verified in tissue-simulating phantoms consisting of Intralipid-20% as a scatterer and either manganese meso-tetra (4-sulfanatophenyl) porphine or intact human erythrocytes as an absorber. Absorption coefficients were recovered with a mean error of 9% and scattering coefficients were recovered with a mean error of 19%, whereas the hemoglobin oxygen saturation was recovered with a mean error of 12%. These results demonstrate the feasibility of optical property recovery for situations in which surface-contact spectroscopy is not a possibility, and where only a single probe can be inserted into the tissue.

We show that terahertz (THz) time-domain spectroscopy (TDS) can be used to characterize the blood. The complex optical constants of blood and its constituents, such as water, plasma, and red blood cells (RBCs), were obtained in the THz frequency region. The volume percentage of RBCs in blood was extracted and compared with the conventional RBC counter results. The THz absorption constants are shown to vary linearly with the RBC concentration in both normal saline and whole blood. The excellent linearity between the THz signal and the RBC concentration was also confirmed in a polyurethane resin tube using a THz imaging method. These results demonstrate that THz-TDS imaging can facilitate the quantitative analysis of blood.

A scientific approach to the formulation of medical and technical requirements (MTRs) for noninvasive spectrophotometric diagnostic devices using optical technologies such as laser Doppler flowmetry and absorption spectroscopy is proposed. The theoretical modeling framework, metrological certification, and testing of these devices are still in the early stages of development. The theoretical estimation of the received signal levels for wavelengths between 514 and 940 nm is highly dependent on the blood volume level in the subject tissue. The proposed approach allows, in particular, the calculation of technical and metrological performance constraints of the instruments, such as the ranges of the sensitivity and power-related signal-to-noise ratios for different spectral channels and different biomedical (biochemical and physiological) parameters. Substantiation of specialized MTRs for the noninvasive spectrophotometric diagnostic devices can enable them to develop to the level of standardized measurement techniques.

The sensitivity analysis indicates that the effective absorption coefficient is most sensitive to the concentration of oxygenated hemoglobin in spectral bands centered at 700 and 960 nm. We find that the highest temporal modulation due to heart function for a thick sample, like an arm, is at 940 nm, a significant shift from 710 nm measured for a finger. The most favorable spectral region for a thick transmission sample, such as a forearm, is the domain defined by intervals [900  nm≤λ 1 ≤1000  nm] and [650  nm≤λ 2 ≤720  nm] . We evaluated five near-infrared light-emitting diodes (LEDs) for their potential applications in oximetry. The LED with peak emission at 930 nm emits well in this spectral region. Here the temporal noise is low, and the effective absorption coefficient is strongly dependent on the concentration of the oxygenated hemoglobin. High-quality saturation results are obtained through the forearm during a short measurement (30 s).

Laser triangulation measurements of Er:YAG and Er,Cr:YSGG laser-ablated volumes in hard dental tissues are made, in order to verify the possible existence of a “hydrokinetic” effect that has been proposed as an alternative to the “subsurface water expansion” mechanism for hard-tissue laser ablation. No evidence of the hydrokinetic effect could be observed under a broad range of tested laser parameters and water cooling conditions. On the contrary, the application of water spray during laser exposure of hard dental material is observed to diminish the laser-ablation efficiency (AE) in comparison with laser exposure under the absence of water spray. Our findings are in agreement with the generally accepted principle of action for erbium laser ablation, which is based on fast subsurface expansion of laser-heated water trapped within the interstitial structure of hard dental tissues. Our measurements also show that the well-known phenomenon of ablation stalling, during a series of consecutive laser pulses, can primarily be attributed to the blocking of laser light by the loosely bound and recondensed desiccated minerals that collect on the tooth surface during and following laser ablation. In addition to the prevention of tooth bulk temperature buildup, a positive function of the water spray that is typically used with erbium dental lasers is to rehydrate these minerals, and thus sustaining the subsurface expansion ablation process. A negative side effect of using a continuous water spray is that the AE gets reduced due to the laser light being partially absorbed in the water-spray particles above the tooth and in the collected water pool on the tooth surface. Finally, no evidence of the influence of the water absorption shift on the hypothesized increase in the AE of the Er,Cr:YSGG wavelength is observed.

This study aimed to assess how the wear that brushing promotes affects CO 2 laser-irradiated enamel microhardness after cariogenic challenge in vitro. Forty fragments measuring 4×4  mm were randomly assigned to four groups according to the enamel surface treatment: G1—control, G2—CO 2 -laser irradiation, G3—brushing, and G4—CO 2 laser irradiation + brushing. A laser device emitting at 10.6 μm was used (power=0.5  W , energy per pulse=0.05  mJ , and frequency=10  kHz ). Specimens belonging to groups G3 and G4 were brushed (80,000 strokes) with a brushing simulator using toothpaste. Next, the samples were challenged with acid: the specimens were immersed in demineralizing and remineralizing solutions for 8 days. The acid resistance of enamel was evaluated by cross-sectional microhardness tests. The area under the curve (KHN×μm ) was calculated. Analysis of variance (ANOVA) one-away and Fisher’s test were performed for the statistical analysis (p<0.05 ). Group G2 specimens (31,185±4706 ) were statistically different from specimens belonging to groups G1 (26,723±2446 ), G3 (28,194±1376 ), and G4 (28,207±2234 ), which were statistically similar. The brushing time used in the present study probably wore the CO 2 -lased enamel, so demineralization could not be prevented in the brushed group.

Pigments of food and beverages could affect dental bleaching efficacy. The aim of this investigation was to evaluate color change and mineral loss of tooth enamel as well as the influence of staining solutions normally used by adolescent patients undergoing home bleaching. Initial hardness and baseline color were measured on enamel blocks. Specimens were divided into five groups (n=5 ): G1 (control) specimens were kept in artificial saliva throughout the experiment (3 weeks); G2 enamel was exposed to 10% carbamide peroxide for 6 h daily, and after this period, the teeth were cleaned and stored in artificial saliva until the next bleaching session; and G3, G4, and G5 received the same treatments as G2, but after bleaching, they were stored for 1 h in cola soft drink, melted chocolate, or red wine, respectively. Mineral loss was obtained by the percentage of hardness reduction, and color change was determined by the difference between the data obtained before and after treatments. Data were subjected to analysis of variance and Fisher’s test (α=0.05 ). G3 and G5 showed higher mineral loss (92.96±5.50 and 94.46±1.00 , respectively) compared to the other groups (p≤0.05 ). G5 showed high-color change (9.34±2.90 ), whereas G1 presented lower color change (2.22±0.44 ) (p≤0.05 ). Acidic drinks cause mineral loss of the enamel, which could modify the surface and reduce staining resistance after bleaching.