A wavelength-coded volume holographic imaging (WC-VHI) endoscope system capable of simultaneous multifocal imaging is presented. The system images light from two depths separated by 100  μm in a tissue sample by using axial chromatic dispersion of a gradient index probe in combination with two light-emitting diode sources and a multiplexed volume hologram to separate the images. This system is different from previous VHI systems in that it uses planar multiplexed gratings and does not require curved holographic gratings. This results in improved lateral imaging resolution from 228.1 to 322.5  lp/mm. This letter describes the design and fabrication of the WC-VHI endoscope and experimental images of hard and soft resolution targets and biological tissue samples to illustrate the performance properties.

To accurately determine sample optical properties using single fiber reflectance spectroscopy (SFR), an absolute calibration of the reflectance is required. We investigated two SFR calibration methods, using a calibrated mirror and using the Fresnel reflection at the fiber tip as a reference. We compared these to commonly used calibration methods, using either Intralipid-20% in combination with Monte Carlo simulations or Spectralon as a reference. The Fresnel reflection method demonstrated the best reproducibility and yielded the most reliable result. We therefore recommend the Fresnel reflection method for the measured absolute reflectance calibration of SFR.

Inadvertent injury to important anatomic structures is a significant risk in minimally invasive surgery (MIS) that potentially requires conversion to an open procedure, which results in increased morbidity and mortality. Surgeons operating minimal-invasively currently do not have an easy-to-use, real-time device to aid in intraoperative identification of important anatomic structures that underlie tissue planes. We demonstrate freehand diffuse optical spectroscopy (DOS) imaging for intraoperatively identifying major underlying veins and arteries. An applicator probe that can be affixed to and detached from an 8-mm laparoscopic instrument has been developed. The 10-mm DOS source–detector separation renders sampling of tissue heterogeneities a few millimeters deep. DOS spectra acquired consecutively during freehand movement of the applicator probe on the tissue surface are displayed as a temporal and spectral image to assist in spatially resolved identification of the underlying structures. Open surgery identifications of the vena cava and aorta underlying peritoneal fat of ∼4  mm in thickness using the applicator probe under room light were demonstrated repeatedly in multiple pigs in vivo.

Myocardial infarction (MI), a cause of significant morbidity and mortality, is typically followed by microstructural alterations where the necrotic myocardium is steadily replaced with a collagen scar. Engineered remodeling of the fibrotic scar via stem cell regeneration has been shown to improve/restore the myocardium function after MI. Nevertheless, the heterogeneous nature of the scar patch may impair the myocardial electrical integrity, leading to the formation of arrhythmogenesis. Radiofrequency ablation (RFA) offers an effective treatment for focal arrhythmias where local heating generated via electric current at specific spots in the myocardium ablate the arrhythmogenic foci. Characterization of these myocardial pathologies (i.e., infarcted, stem cell regenerated, and RFA-ablated myocardial tissues) is of potential clinical importance. Optical polarimetry, the use of light to map and characterize the polarization signatures of a sample, has emerged as a powerful imaging tool for structural characterization of myocardial tissues, exploiting the underlying highly fibrous tissue nature. This study aims to review the recent progress in optical polarimetry pertaining to the characterization of myocardial pathologies while describing the underlying biological rationales that give rise to the optical imaging contrast in various pathologies of the myocardium. Future possibilities of and challenges to optical polarimetry in cardiac imaging clinics are also discussed.

Frequency-domain photon migration (FDPM) uses modulated laser light to measure the bulk optical properties of turbid media and is increasingly applied for noninvasive functional medical imaging in the near-infrared. Although semiconductor edge-emitting laser diodes have been traditionally used as miniature light sources for this application, we show that vertical-cavity surface-emitting lasers (VCSELs) exhibit output power and modulation performance characteristics suitable for FDPM measurements of tissue optical properties at modulation frequencies exceeding 1 GHz. We also show that an array of multiple VCSEL devices can be coherently modulated at frequencies suitable for FDPM and can improve optical power. In addition, their small size and simple packaging make them an attractive choice as components in wearable sensors and clinical FDPM-based optical spectroscopy systems. We demonstrate the benefits of VCSEL technology by fabricating and testing a unique, compact VCSEL-based optical probe with an integrated avalanche photodiode. We demonstrate sensitivity of the VCSEL-based probe to subcutaneous tissue hemodynamics that was induced during an arterial cuff occlusion of the upper arm in a human subject.

A dual-modal approach using Raman spectroscopy and optical pH sensing was investigated to discriminate between normal and cancerous tissues. Raman spectroscopy has demonstrated the potential for in vivo cancer detection. However, Raman spectroscopy has suffered from strong fluorescence background of biological samples and subtle spectral differences between normal and disease tissues. To overcome those issues, pH sensing is adopted to Raman spectroscopy as a dual-modal approach. Based on the fact that the pH level in cancerous tissues is lower than that in normal tissues due to insufficient vasculature formation, the dual-modal approach combining the chemical information of Raman spectrum and the metabolic information of pH level can improve the specificity of cancer diagnosis. From human breast tissue samples, Raman spectra and pH levels are measured using fiber-optic-based Raman and pH probes, respectively. The pH sensing is based on the dependence of pH level on optical transmission spectrum. Multivariate statistical analysis is performed to evaluate the classification capability of the dual-modal method. The analytical results show that the dual-modal method based on Raman spectroscopy and optical pH sensing can improve the performance of cancer classification.

This is a study motivated by the need to develop a needle-free device for eliminating major global healthcare problems caused by needles. The generation of liquid jets by means of a continuous-wave laser, focused into a light absorbing solution, was studied with the aim of developing a portable and affordable jet injector. We designed and fabricated glass microfluidic devices, which consist of a chamber where thermocavitation is created and a tapered channel. The growth of a vapor bubble displaces and expels the liquid through the channel as a fast traveling jet. Different parameters were varied with the purpose of increasing the jet velocity. The velocity increases with smaller channel diameters and taper ratios, whereas larger chambers significantly reduce the jet speed. It was found that the initial position of the liquid–air meniscus interface and its dynamics contribute to increased jet velocities. A maximum velocity of 94±3  m/s for a channel diameter of D=120  μm, taper ratio n=0.25, and chamber length E=200  μm was achieved. Finally, agarose gel-based skin phantoms were used to demonstrate the potential of our devices to penetrate the skin. The maximum penetration depth achieved was ∼1  mm, which is sufficient to penetrate the stratum corneum and for most medical applications. A meta-analysis shows that larger injection volumes will be required as a next step to medical relevance for laser-induced jet injection techniques in general.

The present study evaluated the capability of saliva analysis combining membrane protein purification with surface-enhanced Raman spectroscopy (SERS) for noninvasive detection of nasopharyngeal carcinoma (NPC). A rapid and convenient protein purification method based on cellulose acetate membrane was developed. A total of 659 high-quality SERS spectra were acquired from purified proteins extracted from the saliva samples of 170 patients with pathologically confirmed NPC and 71 healthy volunteers. Spectral analysis of those saliva protein SERS spectra revealed specific changes in some biochemical compositions, which were possibly associated with NPC transformation. Furthermore, principal component analysis combined with linear discriminant analysis (PCA-LDA) was utilized to analyze and classify the saliva protein SERS spectra from NPC and healthy subjects. Diagnostic sensitivity of 70.7%, specificity of 70.3%, and diagnostic accuracy of 70.5% could be achieved by PCA-LDA for NPC identification. These results show that this assay based on saliva protein SERS analysis holds promising potential for developing a rapid, noninvasive, and convenient clinical tool for NPC screening.

We report the application of a hidden Markov model (HMM) on multifractal tissue optical properties derived via the Born approximation-based inverse light scattering method for effective discrimination of precancerous human cervical tissue sites from the normal ones. Two global fractal parameters, generalized Hurst exponent and the corresponding singularity spectrum width, computed by multifractal detrended fluctuation analysis (MFDFA), are used here as potential biomarkers. We develop a methodology that makes use of these multifractal parameters by integrating with different statistical classifiers like the HMM and support vector machine (SVM). It is shown that the MFDFA-HMM integrated model achieves significantly better discrimination between normal and different grades of cancer as compared to the MFDFA-SVM integrated model.

Shape variations of red blood cells (RBCs) are known to occur upon exposure to various drugs or under diseased conditions. The commonly observed discocytic RBCs can be transformed to echinocytic or stomatocytic shape under such conditions. Raman spectra of the three major shape variations, namely discocyte, echinocyte, and stomatocyte, of RBCs were studied while subjecting the cells to oxygenated and deoxygenated conditions. Analysis of the recorded spectra suggests an increased level of hemoglobin (Hb)–oxygen affinity for the echinocytes. Also, some level of Hb degradation could be noticed for the deoxygenated echinocytes. The effects may arise from a reduced level of intracellular adenosine triphosphate in echinocytic cells and an increased fraction of submembrane Hb.

Optical techniques are effective tools for diagnostic applications in medicine and are particularly attractive for the noninvasive analysis of biological tissues and fluids in vivo. Noninvasive examinations of substances via a fiber optic probe need to consider the optical properties of biological tissues obstructing the optical path. This applies to the analysis of the human perilymph, which is located behind the round window membrane. The composition of this inner ear liquid is directly correlated to inner ear hearing loss. In this work, experimental methods for studying the optical properties of the human round window membrane ex vivo are presented. For the first time, a comprehensive investigation of this tissue is performed, including optical transmission, forward scattering, and Raman scattering. The results obtained suggest the application of visible wavelengths ( >400  nm) for investigating the perilymph behind the round window membrane in future.

It is widely accepted that cervical screening has significantly reduced the incidence of cervical cancer worldwide. The primary screening test for cervical cancer is the Papanicolaou (Pap) test, which has extremely variable specificity and sensitivity. There is an unmet clinical need for methods to aid clinicians in the early detection of cervical precancer. Raman spectroscopy is a label-free objective method that can provide a biochemical fingerprint of a given sample. Compared with studies on infrared spectroscopy, relatively few Raman spectroscopy studies have been carried out to date on cervical cytology. The aim of this study was to define the Raman spectral signatures of cervical exfoliated cells present in liquid-based cytology Pap test specimens and to compare the signature of high-grade dysplastic cells to each of the normal cell types. Raman spectra were recorded from single exfoliated cells and subjected to multivariate statistical analysis. The study demonstrated that Raman spectroscopy can identify biochemical signatures associated with the most common cell types seen in liquid-based cytology samples; superficial, intermediate, and parabasal cells. In addition, biochemical changes associated with high-grade dysplasia could be identified suggesting that Raman spectroscopy could be used to aid current cervical screening tests.

We report the results of characterization of red blood cell (RBC) structure and its dynamics with nanometric sensitivity using transport of intensity equation microscopy (TIEM). Conventional transport of intensity technique requires three intensity images and hence is not suitable for studying real-time dynamics of live biological samples. However, assuming the sample to be homogeneous, phase retrieval using transport of intensity equation has been demonstrated with single defocused measurement with x-rays. We adopt this technique for quantitative phase light microscopy of homogenous cells like RBCs. The main merits of this technique are its simplicity, cost-effectiveness, and ease of implementation on a conventional microscope. The phase information can be easily merged with regular bright-field and fluorescence images to provide multidimensional (three-dimensional spatial and temporal) information without any extra complexity in the setup. The phase measurement from the TIEM has been characterized using polymeric microbeads and the noise stability of the system has been analyzed. We explore the structure and real-time dynamics of RBCs and the subdomain membrane fluctuations using this technique.

Transbronchial needle aspiration (TBNA) of small lesions or lymph nodes in the lung may result in nondiagnostic tissue samples. We demonstrate the integration of an optical coherence tomography (OCT) probe into a 19-gauge flexible needle for lung tissue aspiration. This probe allows simultaneous visualization and aspiration of the tissue. By eliminating the need for insertion and withdrawal of a separate imaging probe, this integrated design minimizes the risk of dislodging the needle from the lesion prior to aspiration and may facilitate more accurate placement of the needle. Results from in situ imaging in a sheep lung show clear distinction between solid tissue and two typical constituents of nondiagnostic samples (adipose and lung parenchyma). Clinical translation of this OCT-guided aspiration needle holds promise for improving the diagnostic yield of TBNA.

The lymphatic system is responsible for fluid homeostasis and immune cell trafficking and has been implicated in several diseases, including obesity, diabetes, and cancer metastasis. Despite its importance, the lack of suitable in vivo imaging techniques has hampered our understanding of the lymphatic system. This is, in part, due to the limited contrast of lymphatic fluids and structures. Photoacoustic imaging, in combination with optically absorbing dyes or nanoparticles, has great potential for noninvasively visualizing the lymphatic vessels deep in tissues. Multispectral photoacoustic imaging is capable of separating the components; however, the slow wavelength switching speed of most laser systems is inadequate for imaging lymphatic pumping without motion artifacts being introduced into the processed images. We investigate two approaches for visualizing lymphatic processes in vivo. First, single-wavelength differential photoacoustic imaging is used to visualize lymphatic pumping in the hindlimb of a mouse in real time. Second, a fast-switching multiwavelength photoacoustic imaging system was used to assess the propulsion profile of dyes through the lymphatics in real time. These approaches may have profound impacts in noninvasively characterizing and investigating the lymphatic system.

In optical imaging, the depth and resolution are limited due to scattering. Unlike light, scattering of ultrasound (US) waves in tissue is negligible. Hybrid imaging methods such as US-modulated optical tomography (UOT) use the advantages of both modalities. UOT tags light by inducing phase change caused by modulating the local index of refraction of the medium. The challenge in UOT is detecting the small signal. The displacement induced by the acoustic radiation force (ARF) is another US effect that can be utilized to tag the light. It induces greater phase change, resulting in a stronger signal. Moreover, the absorbed acoustic energy generates heat, resulting in change in the index of refraction and a strong phase change. The speckle pattern is governed by the phase of the interfering scattered waves; hence, speckle pattern analysis can obtain information about displacement and temperature changes. We have presented a model to simulate the insonation processes. Simulation results based on fixed-particle Monte Carlo and experimental results show that the signal acquired by utilizing ARF is stronger compared to UOT. The introduced mean irradiance change (MIC) signal reveals both thermal and mechanical effects of the focused US beam in different timescales. Simulation results suggest that variation in the MIC signal can be used to generate a displacement image of the medium.

Laser speckle-based techniques are frequently used to assess microcirculatory blood flow. Perfusion estimates are calculated either by analyzing the speckle fluctuations over time as in laser Doppler flowmetry (LDF), or by analyzing the speckle contrast as in laser speckle contrast imaging (LSCI). The perfusion estimates depend on the amount of blood and its speed distribution. However, the perfusion estimates are commonly given in arbitrary units as they are nonlinear and depend on the magnitude and the spatial distribution of the optical properties in the tissue under investigation. We describe how the spatial confinement of blood to vessels, called the vessel packaging effect, can be modeled in LDF and LSCI, which affect the Doppler power spectra and speckle contrast, and the underlying bio-optical mechanisms for these effects. As an example, the perfusion estimate is reduced by 25% for LDF and often more than 50% for LSCI when blood is located in vessels with an average diameter of 40  μm, instead of being homogeneously distributed within the tissue. This significant effect can be compensated for only with knowledge of the average diameter of the vessels in the tissue.

Rectal cancer is treated with preoperative radiotherapy (RT) to downstage the tumor, reduce local recurrence, and improve patient survival. Still, the treatment outcome varies significantly and new biomarkers are desired. Collagen I (Col-I) is a potential biomarker, which can be visualized label-free by second harmonic generation (SHG). Here, we used SHG to identify Col-I changes induced by RT in surgical tissue, with the aim to evaluate the clinical significance of RT-induced Col-I changes. First, we established a procedure for quantitative evaluation of Col-I by SHG in CDX2-stained tissue sections. Next, we evaluated Col-I properties in material from 31 non-RT and 29 RT rectal cancer patients. We discovered that the Col-I intensity and anisotropy were higher in the tumor invasive margin than in the inner tumor and normal mucosa, and RT increased and decreased the intensity in inner tumor and normal mucosa, respectively. Furthermore, higher Col-I intensity in the inner tumor was related to increased distant recurrence in the non-RT group but to longer survival in the RT group. In conclusion, we present a new application of SHG for quantitative analysis of Col-I in surgical material, and the first data suggest Col-I intensity as a putative prognostic biomarker in rectal cancer.

A holographic method for high-speed, noncontact photoacoustic tomography is introduced and evaluated. Relative changes of the object’s topography, induced by the impact of thermoelastic pressure waves, were determined at nanometer sensitivity without physical contact. The object’s surface was illuminated with nanosecond laser pulses and imaged with a high-speed CMOS camera. From two interferograms measured before and after excitation of the acoustic wave, surface displacement was calculated and then used as the basis for a tomographic reconstruction of the initial pressure caused by optical absorption. The holographic detection scheme enables variable sampling rates of the photoacoustic signal of up to 50 MHz. The total acquisition times for complete volumes with 230 MVoxel is far below 1 s. Measurements of silicone and porcine skin tissue phantoms with embedded artificial absorbers, which served as a model for human subcutaneous vascular networks, were possible. Three-dimensional reconstructions of the absorbing structures show details with a diameter of 310  μm up to a depth of 2.5 mm. Theoretical limitations and the experimental sensitivity, as well as the potential for in vivo imaging depending on the detection repetition rate, are analyzed and discussed.

Ramification of blood circulation is relevant in a number of physiological and pathological conditions. The oxygen exchange occurs largely in the capillary bed, and the cancer progression is closely linked to the angiogenesis around the tumor mass. Optical microscopy has made impressive improvements in in vivo imaging and dynamic studies based on correlation analysis of time stacks of images. Here, we develop and test advanced methods that allow mapping the flow fields in branched vessel networks at the resolution of 10 to 20  μm. The methods, based on the application of spatiotemporal image correlation spectroscopy and its extension to cross-correlation analysis, are applied here to the case of early stage embryos of zebrafish.

Red blood cell (RBC) anomalies are significant symptoms for identification of health disorders and several blood diseases, which involve the modification of the parameters and biophysical characteristics of such cells. The aim of this study is to measure the three-dimensional phase information of healthy RBCs and their parameters, such as cell diameter, thickness, and hemoglobin (Hb) content, using Talbot-projected fringes. The Talbot image of linear grating is projected onto an RBC slide. The deformed grating lines due to the shape and refractive index of RBCs are recorded by a CCD camera through a 20× microscope objective. Hilbert transform is used to extract the phase image from the deformed projected grating lines. Experimentally calculated values of diameter ( 8.2  μm), thickness ( 2.7  μm), and Hb content ( 28.7  pg/cell) are well within the limits available in the literature. The proposed system is robust and user-friendly and performs the imaging of RBCs with high axial and lateral resolution ( 2.19  μm).

Study of metabolic changes during epithelial–mesenchymal transition (EMT) of cancer cells is important for basic understanding and therapeutic management of cancer progression. We here used metabolic labeling and stimulated Raman scattering (SRS) microscopy, a strategy of bioorthogonal chemical imaging, to directly visualize changes in anabolic metabolism during cancer EMT at a single-cell level. MCF-7 breast cancer cell is employed as a model system. Four types of metabolites (amino acids, glucose, fatty acids, and choline) are labeled with either deuterium or alkyne ( C≡C) tag. Their intracellular incorporations into MCF-7 cells before or after EMT are visualized by SRS imaging targeted at the signature vibration frequency of C-D or C≡C bonds. Overall, after EMT, anabolism of amino acids, glucose, and choline is less active, reflecting slower protein and membrane synthesis in mesenchymal cells. Interestingly, we also observed less incorporation of glucose and palmitate acids into membrane lipids, but more of them into lipid droplets in mesenchymal cells. This result indicates that, although mesenchymal cells synthesize fewer membrane lipids, they are actively storing energy into lipid droplets, either through de novo lipogenesis from glucose or direct scavenging of exogenous free fatty acids. Hence, metabolic labeling coupled with SRS can be a straightforward method in imaging cancer metabolism.

Photoacoustic tomography (PAT) is emerging as a potentially important aid for breast cancer detection. Well-validated tissue-simulating phantoms are needed for objective, quantitative, and physically realistic testing for system development. Prior reported PAT phantoms with homogenous structures do not incorporate the irregular layered structure of breast tissue. To assess the impact of this simplification, we design and construct two-layer breast phantoms incorporating vessel-simulating inclusions and realistic undulations at the fat/fibroglandular tissue interface. The phantoms are composed of custom poly(vinyl chloride) plastisol formulations mimicking the acoustic properties of two breast tissue types and tissue-relevant similar optical properties. Resulting PAT images demonstrate that in tissue with acoustic heterogeneity, lateral size of imaging targets is sensitive to the choice of sound speed in image reconstruction. The undulating boundary can further degrade a target’s lateral size due to sound speed variation in tissue and refraction of sound waves at the interface. The extent of this degradation is also influenced by the geometric relationship between an absorber and the boundary. Results indicate that homogeneous phantom matrixes may underestimate the degradation of PAT image quality in breast tissue, whereas heterogeneous phantoms can provide more realistic testing through improved reproduction of spatial variations in physical properties.

Coronary heart disease (the presence of coronary atherosclerotic plaques) is a significant health problem in the industrialized world. A clinical method to accurately visualize and characterize atherosclerotic plaques is needed. Intravascular photoacoustic (IVPA) imaging is being developed to fill this role, but questions remain regarding optimal imaging wavelengths. We utilized a Monte Carlo optical model to simulate IVPA excitation in coronary tissues, identifying optimal wavelengths for plaque characterization. Near-infrared wavelengths ( ≤1800  nm) were simulated, and single- and dual-wavelength data were analyzed for accuracy of plaque characterization. Results indicate light penetration is best in the range of 1050 to 1370 nm, where 5% residual fluence can be achieved at clinically relevant depths of ≥2  mm in arteries. Across the arterial wall, fluence may vary by over 10-fold, confounding plaque characterization. For single-wavelength results, plaque segmentation accuracy peaked at 1210 and 1720 nm, though correlation was poor ( <0.13). Dual-wavelength analysis proved promising, with 1210 nm as the most successful primary wavelength ( ≈1.0). Results suggest that, without flushing the luminal blood, a primary and secondary wavelength near 1210 and 1350 nm, respectively, may offer the best implementation of dual-wavelength IVPA imaging. These findings could guide the development of a cost-effective clinical system by highlighting optimal wavelengths and improving plaque characterization.

Pain diagnosis for nonverbal patients represents a challenge in clinical settings. Neuroimaging methods, such as functional magnetic resonance imaging and functional near-infrared spectroscopy (fNIRS), have shown promising results to assess neuronal function in response to nociception and pain. Recent studies suggest that neuroimaging in conjunction with machine learning models can be used to predict different cognitive tasks. The aim of this study is to expand previous studies by exploring the classification of fNIRS signals (oxyhaemoglobin) according to temperature level (cold and hot) and corresponding pain intensity (low and high) using machine learning models. Toward this aim, we used the quantitative sensory testing to determine pain threshold and pain tolerance to cold and heat in 18 healthy subjects (three females), mean age±standard deviation ( 31.9±5.5). The classification model is based on the bag-of-words approach, a histogram representation used in document classification based on the frequencies of extracted words and adapted for time series; two learning algorithms were used separately, K-nearest neighbor (K-NN) and support vector machines (SVM). A comparison between two sets of fNIRS channels was also made in the classification task, all 24 channels and 8 channels from the somatosensory region defined as our region of interest (RoI). The results showed that K-NN obtained slightly better results (92.08%) than SVM (91.25%) using the 24 channels; however, the performance slightly dropped using only channels from the RoI with K-NN (91.53%) and SVM (90.83%). These results indicate potential applications of fNIRS in the development of a physiologically based diagnosis of human pain that would benefit vulnerable patients who cannot self-report pain.

Colorectal surgery is the standard treatment for patients with colorectal cancer. To overcome two of the main challenges, the circumferential resection margin and postoperative complications, real-time tissue assessment could be of great benefit during surgery. In this ex vivo study, diffuse reflectance spectroscopy (DRS) was used to differentiate tumor tissue from healthy surrounding tissues in patients with colorectal neoplasia. DRS spectra were obtained from tumor tissue, healthy colon, or rectal wall and fat tissue, for every patient. Data were randomly divided into training (80%) and test (20%) sets. After spectral band selection, the spectra were classified using a quadratic classifier and a linear support vector machine. Of the 38 included patients, 36 had colorectal cancer and 2 had an adenoma. When the classifiers were applied to the test set, colorectal cancer could be discriminated from healthy tissue with an overall accuracy of 0.95 ( ±0.03). This study demonstrates the possibility to separate colorectal cancer from healthy surrounding tissue by applying DRS. High classification accuracies were obtained both in homogeneous and inhomogeneous tissues. This is a fundamental step toward the development of a tool for real-time in vivo tissue assessment during colorectal surgery.

Plastic embedding is widely applied in light microscopy analyses. Previous studies have shown that embedding agents and related techniques can greatly affect the quality of biological tissue embedding and fluorescent imaging. Specifically, it is difficult to preserve endogenous fluorescence using currently available acidic commercial embedding resins and related embedding techniques directly. Here, we developed a neutral embedding resin that improved the green fluorescent protein (GFP), yellow fluorescent protein (YFP), and DsRed fluorescent intensity without adjusting the pH value of monomers or reactivating fluorescence in lye. The embedding resin had a high degree of polymerization, and its fluorescence preservation ratios for GFP, YFP, and DsRed were 126.5%, 155.8%, and 218.4%, respectively.

To advance the practical application of optical coherence tomography (OCT) in the field of biomedical imaging, the imaging depth must be extended without sacrificing resolution while maintaining sufficient sensitivity. However, there is an inherent trade-off between lateral resolution and depth of field (DOF) in OCT. To address this shortcoming, this article proposes a multifocal Bessel beam spectral-domain optical coherence tomography (MBSDOCT) capable of increasing the DOF with unchanged lateral resolution and a high signal-to-noise ratio. The proposed technique is demonstrated by simulation and experiment. A three-focal MBSDOCT with an axicon lens theoretically achieved a DOF of ∼6  mm with a lateral resolution of ∼13  μm. In imaging experiments performed on the acinar cells of orange tissue, a measured DOF of ∼4  mm was demonstrated with a sensitivity penalty of ∼18.1  dB, relative to the Gaussian beam spectral-domain OCT, with a 9-mW light source.

Lung cancer is the most prevalent type of cancer and the leading cause of cancer-related deaths worldwide. Coherent anti-Stokes Raman scattering (CARS) is capable of providing cellular-level images and resolving pathologically related features on human lung tissues. However, conventional means of analyzing CARS images requires extensive image processing, feature engineering, and human intervention. This study demonstrates the feasibility of applying a deep learning algorithm to automatically differentiate normal and cancerous lung tissue images acquired by CARS. We leverage the features learned by pretrained deep neural networks and retrain the model using CARS images as the input. We achieve 89.2% accuracy in classifying normal, small-cell carcinoma, adenocarcinoma, and squamous cell carcinoma lung images. This computational method is a step toward on-the-spot diagnosis of lung cancer and can be further strengthened by the efforts aimed at miniaturizing the CARS technique for fiber-based microendoscopic imaging.

We present noninvasive, three-dimensional, depth-resolved imaging of human retinal and choroidal blood circulation with a swept-source optical coherence tomography (OCT) system at 1065-nm center wavelength. Motion contrast OCT imaging was performed with the phase-variance OCT angiography method. A Fourier-domain mode-locked light source was used to enable an imaging rate of 1.7 MHz. We experimentally demonstrate the challenges and advantages of wide-field OCT angiography (OCTA). In the discussion, we consider acquisition time, scanning area, scanning density, and their influence on visualization of selected features of the retinal and choroidal vascular networks. The OCTA imaging was performed with a field of view of 16 deg ( 5  mm×5  mm) and 30 deg ( 9  mm×9  mm). Data were presented in en face projections generated from single volumes and in en face projection mosaics generated from up to 4 datasets. OCTA imaging at 1.7 MHz A-scan rate was compared with results obtained from a commercial OCTA instrument and with conventional ophthalmic diagnostic methods: fundus photography, fluorescein, and indocyanine green angiography. Comparison of images obtained from all methods is demonstrated using the same eye of a healthy volunteer. For example, imaging of retinal pathology is presented in three cases of advanced age-related macular degeneration.

A label-free technique for the detection of triacylglycerides by a localized surface plasmon resonance (LSPR)–based biosensor is demonstrated. An LSPR-based fiber-optic sensor probe is fabricated by immobilizing lipase enzyme on silver nanoparticles (Ag-NPs) coated on an unclad segment of a plastic clad optical fiber. The size and shape of nanoparticles were characterized by high-resolution transmission electron microscopy and UV–visible spectroscopy. The peak absorbance wavelength changes with concentration of triacylglycerides surrounding the sensor probe, and sensitivity is estimated from shift in the peak absorbance wavelength as a function of concentration. The fabricated sensor was characterized for the concentration of triacylglyceride solution in the range 0 to 7 mM. The sensor shows the best sensitivity at a temperature of 37°C and pH 7.4 of the triacylglycerides emulsion with a response time of 40 s. A sensitivity of 28.5  nm/mM of triacylglyceride solution is obtained with a limit of detection of 0.016 mM in the entire range of triacylglycerides. This compact biosensor shows good selectivity, stability, and reproducibility in the entire physiological range of triacylglycerides and is well-suited to real-time online monitoring and remote sensing.

The maximum level of voluntary bite force, which results from the combined action of muscle of mastication, joints, and teeth, i.e., craniomandibular structure, is considered as one of the major indicators for the functional state of the masticatory system. Measurement of voluntary bite force provides useful data for the jaw muscle function and activity along with assessment of prosthetics. This study proposes an in vivo methodology for the dynamic measurement of bite force employing a fiber Bragg grating (FBG) sensor known as bite force measurement device (BFMD). The BFMD developed is a noninvasive intraoral device, which transduces the bite force exerted at the occlusal surface into strain variations on a metal plate. These strain variations are acquired by the FBG sensor bonded over it. The BFMD developed facilitates adjustment of the distance between the biting platform, which is essential to capture the maximum voluntary bite force at three different positions of teeth, namely incisor, premolar, and molar sites. The clinically relevant bite forces are measured at incisor, molar, and premolar position and have been compared against each other. Furthermore, the bite forces measured with all subjects are segregated according to gender and also compared against each other.

This erratum corrects two errors in the original paper.