We propose an algorithm to compensate for the refractive index error in the optical coherence tomography (OCT) images of multilayer tissues, such as skin. The performance of the proposed method has been evaluated on one- and two-layer solid phantoms, as well as the skin of rat paw.

In recent years, the use of through-focus (TF) or volumetric type of optical imaging has gained momentum in several areas such as biological imaging, microscopy, adaptive optics, material processing, optical data storage, and optical inspection. We provide a review of basic TF optical methods highlighting their design, major unique characteristics, and application space.

A guest editorial to introduce the Special Section on Selected Topics in Biophotonics: Optogenetics and Label-Free Optical Spectroscopy

An algorithm that selects optimal wavelengths for spectral fitting of diffuse light reflectance spectra using a nonnegative least squares method is presented. Oxyhemoglobin, deoxyhemoglobin, and water are considered representative absorbers, but the approach is not constrained or limited by absorber selection provided native basis spectra are available. The method removes wavelengths iteratively from a scattering-modulated absorption matrix by maximizing the product of its singular values and offers considerable improvements over previously published wavelength selection schemes. Resulting wavelength selections are valid for a broad range of optical properties and yield lower RMS errors than other wavelength combinations. The method is easily modified and broadly applicable to tissue optical spectroscopy. Adaptation of the algorithm to select optimal light-emitting diodes for fitting blood is described.

As dental caries is one of the most common diseases, the early and noninvasive detection of carious lesions plays an important role in public health care. Optical coherence tomography (OCT) with its ability of depth-resolved, high-resolution, noninvasive, fast imaging has been previously recognized as a promising tool in dentistry. Additionally, polarization sensitive imaging provides quantitative measures on the birefringent tissue properties and can be utilized for imaging dental tissue, especially enamel and dentin. By imaging three exemplary tooth samples ex vivo with proximal white spot, brown spot, and cavity, we show that the combination of polarization sensitive OCT and the degree of polarization uniformity (DOPU) algorithm is a promising approach for the detection of proximal carious lesions due to the depolarization contrast of demineralized tissue. Furthermore, we investigate different sizes of the DOPU evaluation kernel on the resulting contrast and conclude a suitable value for this application. We propose that DOPU provides an easy to interpret image representation and appropriate contrast for possible future screening applications in early caries diagnostics.

As limited data photoacoustic tomographic image reconstruction problem is known to be ill-posed, the iterative reconstruction methods were proven to be effective in terms of providing good quality initial pressure distribution. Often, these iterative methods require a large number of iterations to converge to a solution, in turn making the image reconstruction procedure computationally inefficient. In this work, two variants of vector polynomial extrapolation techniques were deployed to accelerate two standard iterative photoacoustic image reconstruction algorithms, including regularized steepest descent and total variation regularization methods. It is shown using numerical and experimental phantom cases that these extrapolation methods that are proposed in this work can provide significant acceleration (as high as 4.7 times) along with added advantage of improving reconstructed image quality.

Brain tissue analysis is highly desired in neurosurgery, such as tumor resection. To guarantee best life quality afterward, exact navigation within the brain during the surgery is essential. So far, no method has been established that perfectly fulfills this need. Optical coherence tomography (OCT) is a promising three-dimensional imaging tool to support neurosurgical resections. We perform a preliminary study toward in vivo brain tumor removal assistance by investigating meningioma, healthy white, and healthy gray matter. For that purpose, we utilized a commercially available OCT device (Thorlabs Callisto) and measured eight samples of meningioma, three samples of healthy white, and two samples of healthy gray matter ex vivo directly after removal. Structural variations of different tissue types, especially meningioma, can already be seen in the raw OCT images. Nevertheless, an automated differentiation approach is desired, so that neurosurgical guidance can be delivered without a-priori knowledge of the surgeon. Therefore, we employ different algorithms to extract texture features and apply pattern recognition methods for their classification. With these postprocessing steps, an accuracy of nearly 98% was found.

In laserosteotomy, it is vital to avoid thermal damage of the surrounding tissue, such as carbonization, since carbonization does not only deteriorate the ablation efficiency but also prolongs the healing process. The state-of-the-art method to avoid carbonization is irrigation systems; however, it is difficult to determine the desired flow rate of the air and cooling water based on previous experiments without online monitoring of the bone surface. Lack of such feedback during the ablation process can cause carbonization in case of a possible error in the irrigation system or slow down the cutting process when irrigating with too much cooling water. The aim of this paper is to examine laser-induced breakdown spectroscopy as a potential tool for autocarbonization detection in laserosteotomy. By monitoring the laser-driven plasma generated during nanosecond pulse ablation of porcine bone samples, carbonization is hypothesized to be detectable. For this, the collected spectra were analyzed based on variation of a specific pair of emission line ratios in both groups of samples: normal and carbonized bone. The results confirmed a high accuracy of over 95% in classifying normal and carbonized bone.

The common way to diagnose hard and soft tissue irregularities in the oral cavity is initially the visual inspection by an experienced dentist followed by further medical examinations, such as radiological imaging and/or histopathological investigation. For the diagnosis of oral hard and soft tissues, the detection of early transformations is mostly hampered by poor visual access, low specificity of the diagnosis techniques, and/or limited feasibility of frequent screenings. Therefore, optical noninvasive diagnosis of oral tissue is promising to improve the accuracy of oral screening. Considering this demand, a rigid handheld endoscopic scanner was developed for optical coherence tomography (OCT). The novelty is the usage of a commercially near-infrared endoscope with fitting optics in combination with an established spectral-domain OCT system of our workgroup. By reaching a high spatial resolution, in vivo images of anterior and especially posterior dental and mucosal tissues were obtained from the oral cavity of two volunteers. The convincing image quality of the endoscopic OCT device is particularly obvious for the imaging of different regions of the human soft palate with highly scattering fibrous layer and capillary network within the lamina propria.

We describe a Monte Carlo model of the mouse torso to optimize illumination of the mouse lung for fluorescence detection of low levels of pulmonary pathogens, specifically Mycobacterium tuberculosis. After validation of the simulation with an internally illuminated optical phantom, the entire mouse torso was simulated to compare external and internal illumination techniques. Measured optical properties of deflated mouse lungs were scaled to mimic the diffusive properties of inflated lungs in vivo. Using the full-torso model, a 2  ×   to 3  ×   improvement in average fluence rate in the lung was seen for dorsal compared with ventral positioning of the mouse with external illumination. The enhancement in average fluence rate in the lung using internal excitation was 40  ×   to 60  ×   over external illumination in the dorsal position. Parameters of the internal fiber optic source were manipulated in the model to guide optimization of the physical system and experimental protocol for internal illumination and whole-body detection of fluorescent mycobacteria in a mouse model of infection.

Biological tissue is a highly scattering medium that prevents deep imaging of light. For medical applications, optical imaging offers a molecular sensitivity that would be beneficial for diagnosing and monitoring of diseases. Acousto-optical tomography has the molecular sensitivity of optical imaging with the resolution of ultrasound and has the potential for deep tissue imaging. Here, we present a theoretical study of a system that combines acousto-optical tomography and slow light spectral filters created using spectral hole burning methods. Using Monte Carlo simulations, a model to obtain the contrast-to-noise ratio (CNR) deep in biological tissue was developed. The simulations show a CNR  >  1 for imaging depths of ∼5  cm in a reflection mode setup, as well as, imaging through ∼12 cm in transmission mode setups. These results are promising and form the basis for future experimental studies.

For more than two decades, Raman spectroscopy has found widespread use in biological and medical applications. The instrumentation and the statistical evaluation procedures have matured, enabling the lengthy transition from ex-vivo demonstration to in-vivo examinations. This transition goes hand-in-hand with many technological developments and tightly bound requirements for a successful implementation in a clinical environment, which are often difficult to assess for novice scientists in the field. This review outlines the required instrumentation and instrumentation parameters, designs, and developments of fiber optic probes for the in-vivo applications in a clinical setting. It aims at providing an overview of contemporary technology and clinical trials and attempts to identify future developments necessary to bring the emerging technology to the clinical end users. A comprehensive overview of in-vivo applications of fiber optic Raman probes to characterize different tissue and disease types is also given.

Models mimicking the endogenous production of protoporphyrin IX (PpIX), as well as its fluorescence, are of high interest for applied and fundamental studies in the fields of cancer detection by fluorescence imaging, photodynamic therapy (PDT), and photobiomodulation (PBM). Here, we present and describe optical properties of the yeast-based models able to produce PpIX endogenously after the administration of 5-aminolevulinic acid (ALA) and/or 2,2’-bipyridyl. As their optical properties have an important impact on the spatial distribution of the fluence rate in these liquid models, their absorption and reduced scattering coefficients were determined to be between 400 and 808 nm for two yeast solutions previously described by our group. These coefficients were derived from measurements of the total reflectance and light penetration depth using a dedicated Monte Carlo simulation. We observed that absorption and scattering coefficients were smaller than those of soft tissues at all wavelengths. This work will enable the production of a low-cost optical phantom loaded with appropriate amounts of light-absorbing and -scattering particles to mimic tumors containing PpIX, offering a useful tool to optimize the spectral and radiometric design of certain cancer photodetection setups.

Surface-enhanced Raman spectroscopy (SERS) of blood plasma on an electrochemically prepared silver surface has been studied as a label-free, noninvasive diagnostic test for colorectal cancer. Indium tin oxide glass substrates were modified with 0.01  mol dm^−  3 silver nitrate using the pulsed double-potentiostatic method. The prepared silver substrates were tested with Rhodamine 6G as a model analyte and the surface with the highest signal enhancement was selected. This silver dendritic surface was used as a diagnostic substrate for SERS measurements of human blood plasma. A group of oncological patients with declared colorectal carcinoma (n  =  15) and the control group of healthy volunteers (n  =  15) were compared. The biomolecular changes in chemical composition in the cancer samples were detected by statistical processing of the resulting SERS spectra. About 94% specificity and 100% sensitivity were achieved for the analysis by the ratio of the SERS peak intensity at 725  cm^−  1 for adenine to the peak intensity at 638  cm^−  1 for tyrosine and 100% specificity and sensitivity by using principal component analysis. This method of SERS diagnostics of colorectal cancer, which does not require the nanoparticle preparation, mixing, and incubation of plasma with a colloidal solution as in conventional tests, is a rapid, inexpensive method, which could be introduced as a primary diagnostic test.

While there are a plethora of in vivo fiber-optic spectroscopic techniques that have demonstrated the ability to detect a number of diseases in research trials with highly trained personnel familiar with the operation of experimental optical technologies, very few techniques show the same level of success in large multicenter trials. To meet the stringent requirements for a viable optical spectroscopy system to be used in a clinical setting, we developed components including an automated calibration tool, optical contact sensor for signal acquisition, and a methodology for real-time in vivo probe calibration correction. The end result is a state-of-the-art medical device that can be realistically used by a physician with spectroscopic fiber-optic probes. We show how the features of this system allow it to have excellent stability measuring two scattering phantoms in a clinical setting by clinical staff with ∼0.5  %   standard deviation over 25 unique measurements on different days. In addition, we show the systems’ ability to overcome many technical obstacles that spectroscopy applications often face such as speckle noise and user variability. While this system has been designed and optimized for our specific application, the system and design concepts are applicable to most in vivo fiber-optic-based spectroscopic techniques.

Förster resonance energy transfer (FRET) continues to be a useful tool to study movement and interaction between proteins within living cells. When FRET as an optical technique is measured with flow cytometry, conformational changes of proteins can be rapidly measured cell-by-cell for the benefit of screening and profiling. We exploit FRET to study the extent of activation of α4β1 integrin dimers expressed on the surface of leukocytes. The stalk-like transmembrane heterodimers when not active lay bent and upon activation extend outward. Integrin extension is determined by changes in the distance of closest approach between an FRET donor and acceptor, bound at the integrin head and cell membrane, respectively. Time-resolved flow cytometry analysis revealed donor emission increases up to 17%, fluorescence lifetime shifts over 1.0 ns during activation, and FRET efficiencies of 37% and 26% corresponding to the inactive and active integrin state, respectively. Last, a graphical phasor analysis, including population clustering, gating, and formation of an FRET trajectory, added precision to a comparative analysis of populations undergoing FRET, partial donor recovery, and complete donor recovery. This work establishes a quantitative cytometric approach for profiling fluorescence donor decay kinetics during integrin conformational changes on a single-cell level.

Photoplethysmography (PPG) is an optical noninvasive technique with the potential for assessing tissue perfusion. The relative time-change in the concentration of oxyhemoglobin and deoxyhemoglobin in the blood can be derived from DC part of the PPG signal. However, the absolute concentration cannot be determined due to the inadequate data on PPG optical paths. The optical path and differential pathlength factor (DPF) for PPG at red (660 nm) and infrared (880 nm) wavelengths were investigated using a heterogeneous Monte Carlo model of the human forearm. Using the simulated DPFs, the absolute time-change in concentrations were determined from PPG signals recorded from the same tissue site. Results were compared with three conditions of approximated DPFs. Results showed the variation of the optical-path and DPF with different wavelengths and source–detector separations. Approximations resulted in significant errors, for example, using NIRS DPF in PPG led to “cross talk” of −0.4297 and 0.060 and an error of 15.16% to 25.18%. Results confirmed the feasibility of using the PPG (DC) for the assessment of tissue perfusion. The study also identified the inappropriateness of the assumption that DPF is independent of wavelength or source–detector separations and set the platform for further studies on investigating optical pathlengths and DPF in PPG.

The optical properties of human brain tumor tissues, including glioblastoma, meningioma, oligodendroglioma, and metastasis, that were classified into “strong,” “vague,” and “unobservable” fluorescence by a neurosurgeon were measured and compared. The optical properties of the tissues were measured with a double integrating sphere and the inverse Monte Carlo technique from 350 to 1000 nm. Using reasons of ex-vivo measurement, the optical properties at around 420 nm were potentially affected by the hemoglobin content in tissues. Significant differences were not observed between the optical properties of the glioblastoma regions with “strong” and “unobservable” fluorescence. Sections of human brain tumor tissue with “strong” and “unobservable” fluorescence were stained with hematoxylin and eosin. The cell densities [mean ± standard deviation (S.D.)] in regions with “strong” and “unobservable” fluorescence were 31  ±  9  ×  10²  per mm² and 12  ±  4  ×  10²  per mm², respectively, which is a statistically significant difference. The higher fluorescence intensity is associated with higher cell density. The difference in cell density modified the scattering coefficient yet it does not lead to significant differences in the reduced scattering coefficient and thus does not affect the propagation of the diffuse fluorescent light. Hence, the false negatives, which mean a brain tumor only shows “unobservable” fluorescence and is hence classified incorrectly as nontumor, in using 5-ALA for detection of human glioblastoma do not result from the differences in optical properties of human brain glioblastoma tissues. Our results suggest that the primary cause of false negatives may be a lack of PpIX or a low accumulation of PpIX.

A light-based processing method to create an amorphous trehalose matrix for the stabilization of proteins is discussed. This method has potential applications in the stabilization of protein-based therapeutics and diagnostics. During light-assisted drying (LAD), proteins suspended in a trehalose solution are dehydrated using near-infrared (NIR) laser light. The goal of this study was to determine processing parameters that resulted in fast processing times and low end moisture contents (EMC), while maintaining the functionality of embedded proteins. We compared the effect of changing processing wavelength, power and resulting sample temperature, and substrate material on the EMC for two NIR laser sources (1064 and 1850 nm). The 1850-nm laser resulted in the lowest EMC (0.03  ±  0.01  gH₂O  /  gDryWeight) after 20 min of processing on glass microfiber paper. This suggests a storage temperature of 68.3°C. We also tested the functionality of a model protein, lysozyme, after LAD processing using a standard assay. LAD showed no significant effect on the functionality of lysozyme when processed at a maximum temperature of ∼44  °  C to an EMC of 0.17  ±  0.06  gH₂O  /  gDryWeight. LAD is a promising technique for forming amorphous trehalose solids that could stabilize proteins at ambient temperatures.

The objective of this study was to evaluate the effects of varying light doses on the viability and cellular activity of osteoblasts, osteocytes, and osteoclasts. A light application device was developed to apply 940-nm wavelength light from light-emitting diodes on three cultured cells, MC3T3-E1, MLO-A5, and RANKL-treated RAW264.7 cells. The doses (energy density) on cells were 0, 1, 5, and 7.5  J  /  cm². The corresponding light power densities at the cell site were 0, 1.67, 8.33, and 12.5  mW  /  cm², respectively, and the duration was 10 min. The results showed that the three cell types respond differently to light and their responses were dose dependent. Low-dose treatment (1  J  /  cm²) enhanced osteoblast proliferation, osteoclast differentiation, and osteoclastic bone resorption activity. Osteocyte proliferation was not affected by both low- and high-dose (5  J  /  cm²) treatments. While 1  J  /  cm² did not affect viability of all three cell types, 5  J  /  cm² significantly decreased viability of osteocytes and osteoclasts. Osteoblast viability was negatively impacted by the higher dose (7.5  J  /  cm²). The findings suggest that optimal doses exist for osteoblast and osteoclast, which can stimulate cell activities, and there is a safe dose range for each type of cell tested.

Optical-resolution photoacoustic microscopy (OR-PAM) has demonstrated fast, label-free volumetric imaging of optical-absorption contrast within the quasiballistic regime of photon scattering. However, the limited numerical aperture of the ultrasonic transducer restricts the detectability of the photoacoustic waves, thus resulting in incomplete reconstructed features. To tackle the limited-view problem, we added an oblique illumination beam to the original coaxial optical-acoustic scheme to provide a complementary detection view. The virtual augmentation of the detection view was validated through numerical simulations and tissue-phantom experiments. More importantly, the combination of top and oblique illumination successfully imaged a mouse brain in vivo down to 1 mm in depth, showing detailed brain vasculature. Of special note, it clearly revealed the diving vessels that were long missing in images from original OR-PAM.

Surgical nerve damage due to difficulty with identification remains a major risk for postsurgical complications and decreased quality of life. Fluorescence-guided surgery offers a means to specifically highlight tissues of interest such as nerves and a number of fluorescence-guided surgical systems are in clinical trial or are approved for clinical use. However, no clinically approved nerve-specific fluorophores exist. In addition, many preclinical nerve-specific fluorophores tend to accumulate in adipose tissue due to the molecular composition similarities between the two tissues, making it challenging to generate a specific nerve signal. To alleviate this difficulty, we have synthesized a library of oxazine fluorophores based on the Nile Red scaffold, with the goal of strong adipose specificity without nerve uptake to facilitate ratiometric imaging. The library was screened for tissue specificity ex vivo and in vivo, enabling quantification of adipose-, nerve- and muscle-specific uptake as well as selection of the best candidate for adipose selectivity without nerve signal. We showed our selected Nile Red fluorophore improved nerve contrast using ratiometric imaging, especially nerve-to-adipose contrast as compared to the parent Nile Red compound or nerve-specific imaging alone. This adipose-specific Nile Red derivative could be used in future fluorescence-guided surgery applications where adipose- or nerve-specific contrast is required.

Thulium fiber laser lithotripsy using smaller optical fibers may enable development of miniature ureteroscopes. Two ureteroscope distal tip prototypes were built and characterized. The first design was 4.5-French (Fr) [1.5-mm outer diameter (OD)], five channel tip, housing 200-μm inner diameter (ID) dedicated central channel for insertion of 100-μm core fibers and four surrounding channels, each with 1.5 Fr (510-μm ID) for instrumentation, irrigation, imaging, and illumination, respectively. The second design was 6.0-Fr (2.0-mm OD), three-dimensional printed tip with larger, hemispherical common working channel and separate detection port integrated with ring lighting. Standard instruments, including optical fibers, guidewires, and stone baskets, were inserted through working channels to demonstrate feasibility. Gravitational and manual pump-assisted saline irrigation rates were measured. Luminous intensity distribution curves (LIDCs) were modeled for both ring and conventional lighting designs. Imaging was conducted using 3000, 6000, and 10,000 pixel, miniature, flexible endoscopes with 0.4-, 0.6-, and 0.9-mm OD, to differentiate between urinary stones and ureter wall, for potential clinical application. The multichannel ureteroscope tip with 1.5-Fr working channel yielded a gravitational saline flow rate of 3.9  ±  0.2  mL  /  min compared to 31.3  ±  0.6  mL  /  min for standard (3.6 Fr) ureteroscope channel. Manual, pump-assisted irrigation increased flow rate to 32.5  ±  3.0  mL  /  min. The 6000 pixel, 0.6-mm OD, flexible endoscope provided a balance of clear differentiation between stones and ureter wall and sufficiently small OD. A ring lighting configuration provided more uniform illumination than conventional cross-lighting geometry as demonstrated by LIDCs. With further development, these miniature ureteroscope tip designs may be integrated into a fully functional ureteroscope to permit ureteral access with minimal trauma and improved patient safety and comfort.

Atrial fibrillation (AF) is the most common cardiac arrhythmia and has high patient morbidity. One of the root causes of AF is initiating triggers from atrial myocardium extending into the pulmonary veins. Visualizing the muscular bundles of myocardial extension is essential to guide the catheter radio-frequency ablation and confirm the curative tissue necrosis thereafter. We applied optical coherence tomography (OCT) for direct visualization of cardial muscle extension in myocardium pulmonary junction. Two perspectives (cross-sectional and en face images) are presented for imaging myocardial extensions. The results demonstrated that cross-sectional images can quickly locate the myocardium pulmonary junction. And en face images provide depth-resolved arrangement information of muscular bundles in the myocardium pulmonary junction. The results indicated that OCT could potentially be used to guide catheter radio-frequency ablation for treatment of AF.

A highly sensitive, modular three-color fluorescence endomicroscopy imaging platform spanning the visible to near-infrared (NIR) range is demonstrated. Light-emitting diodes (LEDs) were sequentially pulsed along with the camera acquisition to provide up to 20 frames per second (fps) three-color imaging performance or 60 fps single color imaging. The system was characterized for bacterial and cellular molecular imaging in ex vivo human lung tissue and for bacterial and indocyanine green imaging in ex vivo perfused sheep lungs. A practical method to reduce background tissue autofluorescence is also proposed. The platform was clinically translated into six patients with pulmonary disease to delineate healthy, cancerous, and fibrotic tissue autofluorescent structures. The instrument is the most broadband clinical endomicroscopy system developed to date (covering visible to the NIR, 500 to 900 nm) and demonstrates significant potential for future clinical utility due to its low cost and modular capability to suit a wide variety of molecular imaging applications.

Correlation mapping optical coherence tomography (cmOCT) is a powerful technique for the imaging of skin microvessels structure, based on the discrimination of the static and dynamic regions of the tissue. Although the suitability of cmOCT to visualize the microcirculation has been proved in humans and animal models, less evidence has been provided about its application to examine functional dynamics. Therefore, the goal of this research was validating the cmOCT method for the investigation into microvascular function and vasomotion. A spectral domain optical coherence tomography (SD-OCT) device was employed to image 90 sequential three-dimensional (3-D) OCT volumes from the forearm of 12 volunteers during a 25-min postocclusive reactive hyperemia (PORH) test. The volumes were processed using cmOCT to generate blood flow maps at selected cutaneous depths. The maps clearly trace flow variations during the PORH response for both capillaries and arterioles/venules microvascular layers. Continuous blood flow signals were reconstructed from cmOCT maps to study vasomotion by applying wavelet transform spectral analysis, which revealed fluctuations of flow during PORH, reflecting the regulation of microvascular tone mediated by endothelial cells and sympathetic nerves. The results clearly demonstrate that cmOCT allows the generation of functional information that may be used for diagnostic applications.

Independent component analysis (ICA) is an unmixing method based on a linear model. It has previously been applied in in vivo multiwavelength photoacoustic imaging studies to unmix the components representing individual chromophores by assuming that they are statistically independent. Numerically simulated and experimentally acquired two-dimensional images of tissue-mimicking phantoms are used to investigate the conditions required for ICA to give accurate estimates of the relative chromophore concentrations. A simple approximate fluence correction was applied to reduce but not completely remove the nonlinear fluence distortion, as might be possible in practice. The results show that ICA is robust against the residual effect of the partially corrected fluence distortion. ICA is shown to provide accurate unmixing of the chromophores when the absorption coefficient is within a certain range of values, where the upper absorption threshold is comparable to the absorption of blood. When the absorption is increased beyond these thresholds, ICA abruptly fails to unmix the chromophores accurately. The ICA approach was compared to a linear spectroscopic inversion (SI) with known absorption spectra. In cases where the mixing matrix with the specific absorption spectra is ill-conditioned, ICA is able to provide accurate unmixing when SI results in large errors.

We report herein the fabrication and performance response of a three-dimensional (3-D) intraoral scan probe based on optical coherence tomography (OCT) that enables 3-D structural and functional diagnoses of the human teeth. The OCT system was configured using a swept-source OCT (SS-OCT) with a center wavelength of 1310 nm. The scan probe was built using an MEMS mirror and an optical collimator. The implemented SS-OCT equipped with the MEMS-based scan probe yielded an axial resolution of 10  μm and a scan range of 8  ×  8  mm². Two-dimensional (2-D) cross-sectional images of the teeth were acquired by the scan probe based on the OCT. The 3-D volume image was acquired by combining a series of 2-D images, which includes internal and structural information of the human teeth. To utilize the OCT system as an intraoral scanner, partially overlapped 3-D volume images were sequentially acquired and stitched. The 3-D stitching was implemented based on an iterative closest point algorithm. The feasibility of the intraoral scan probe is demonstrated based on its ability to image and characterize the structure and function of the human teeth.

Fluorescence imaging has been used to evaluate the physiological features of renal ischemia in animal model. However, the fluorophore distribution details of the ischemia model could not be fully represented due to the limited dynamic range of the charged-couple device. A high-dynamic-range (HDR) strategy was adopted in renal ischemia fluorescence imaging, both ex vivo and in vivo. The HDR strategy successfully combined ischemia relevant biological features that could only be captured with different exposure times, and then presented these features in the HDR results. The HDR results effectively highlighted the renal ischemic areas with relatively better perfusion and diminished the saturation that resulted from long exposure time. The relative fluorescence intensities of the ischemic kidneys and the image entropy values were significantly higher in the HDR images than in the original images, therefore enhancing the visualization of the renal ischemia model. The results suggest that HDR could serve as a postprocessing strategy to enhance the assessment of in vivo renal ischemia, and HDR fluorescence molecular imaging could be a valuable imaging tool for future studies of clinical ischemia detection and evaluation.

A challenging issue in photoacoustic biomedical imaging is to take into account the presence of dispersive acoustic media, since these are prone to induce amplitude attenuation and scattering of the photoacoustic frequency components. These perturbations are largely the cause for which the photoacoustic tomographic image reconstruction from projections lacks a plane-wave transport formalism. Attending this problem, we further develop an analytic formalism of the transport and its numerical implementation accounting for dispersive acoustic media. We differentiate three variations of an acoustically perturbing media. Our object of interest is a numerical description of the light absorption map of a coronal human breast image. Then, we analyze conditions for which the propagation of photoacoustic perturbations can obey the generalized Heaviside telegraph equation. In addition, we provide a study of the causality consistency of the wave propagation models. We observe transport implications due to the presence of dispersive acoustic media and derive model adjustments that include attenuation and diffusion approximations within the two-dimensional forward problem. Next, we restore the inverse problem description with the deduced perturbation components. Finally, we solve the nonlinear inverse problem with a numerical strategy for a filtered backprojection reconstruction. At a stage prior to the image reconstruction, we compensate for the effect of acoustic attenuation and diffusion to calculate the inversions of the wave perturbations located within the projections. In this way, we manage to significantly reduce reconstruction artifacts. In consequence, we prevent the use of some additional image processing of noise reduction. We demonstrate a feasible strategy on how to solve the stated nonlinear inverse problem of photoacoustic tomography accounting for dispersive acoustic media. In particular, we emphasize efforts to achieve an analytical description, and thus an algorithm is placed, for imaged sound perturbations to be cleaned from acoustic scattering in a simplified manner.

Spatial frequency domain imaging (SFDI) is a widefield, noncontact, and label-free imaging modality that is currently being explored as a new tool for longitudinal tracking of cancer therapies in the preclinical setting. We describe a two-layer look-up-table (LUT) inversion algorithm for SFDI that better accounts for the skin (top layer) and tumor (bottom layer) tissue geometry in subcutaneous tumor models. Monte Carlo (MC) simulations were conducted natively in the spatial frequency domain, avoiding discretization errors associated with Fourier or Hankel transforms of conventional MC simulation results. The two-layer LUT was validated using two-layer tissue mimicking optical phantoms, in which the optical property extractions of the bottom (tumor) layer were determined to be within 20% and 11% of the true values for μ_a and μs′, respectively. A sensitivity analysis was conducted to evaluate how imperfect top layer estimates affect bottom-layer optical property extractions. Finally, the two-layer LUT was used to reanalyze a prior longitudinal data set, which revealed larger therapy-induced changes in optical scattering and a more hypoxic tumor environment compared to the homogeneous LUT. The two-layer LUT described here improves the accuracy of subcutaneous tumor imaging, and the general methodology can be applied for arbitrary multilayer SFDI applications.

Given known correlations between vascular health and cognitive impairment, the development of tools to image microvasculature in the whole brain could help investigate these correlations. We explore the feasibility of using an automated serial two-photon microscope to image fluorescent gelatin-filled whole rodent brains in three-dimensions (3-D) with the goal of carrying group studies. Vascular density (VD) was computed using automatic segmentation combined with coregistration techniques to build a group-level vascular metric in the whole brain. Focusing on the medial prefrontal cortex, cerebral cortex, the olfactory bulb, and the hippocampal formation, we compared the VD of three age groups (2-, 4.5-, and 8-months-old), for both wild type mice and a transgenic model (APP/PS1) with pathology resembling Alzheimer’s disease (AD). We report a general loss of VD caused by the aging process with a small VD increase in the diseased animals in the somatomotor and somatosensory cortical regions and the olfactory bulb, partly supported by MRI perfusion data. This study supports previous observations that AD transgenic mice show a higher VD in specific regions compared with WT mice during the early and late stages of the disease (4.5 to 8 months), extending results to whole brain mapping.

This publisher’s note explains the change in citation data for 3 articles.