There would be clinical value in a miniature optical-sectioning microscope to enable in vivo interrogation of tissues as a real-time and noninvasive alternative to gold-standard histopathology for early disease detection and surgical guidance. To address this need, a reflectance-based handheld line-scanned dual-axis confocal microscope was developed and fully packaged for label-free imaging of human skin and oral mucosa. This device can collect images at >15  frames  /  s with an optical-sectioning thickness and lateral resolution of 1.7 and 1.1  μm, respectively. Incorporation of a sterile lens cap design enables pressure-sensitive adjustment of the imaging depth by the user during clinical use. In vivo human images and videos are obtained to demonstrate the capabilities of this high-speed optical-sectioning microscopy device.

Cerebral endovascular neurosurgery has transformed the way we manage cerebrovascular disease. Several landmark trials have demonstrated the effectiveness of endovascular techniques leading to continued technological development and applications for various diseases. The utilization of optical technologies and devices is already underway in the field of endovascular neurosurgery. We discuss the contemporary paradigms, challenges, and current optical applications for the most common cerebrovascular diseases: carotid atherosclerotic disease, cerebral aneurysms, intracranial atherosclerosis, and dural arteriovenous fistulas. We also describe needs-based opportunities for future optical applications, with the goal of providing researchers a sense of where we feel optical technologies could impact the way we manage cerebral disease.

Calibration, quantification, and standardization of the polarimetric instrumentation, as well as interpretation and understanding of the obtained data, require the development and use of well-calibrated phantoms and standards. We reviewed the status of tissue phantoms for a variety of applications in polarimetry; more than 500 papers are considered. We divided the phantoms into five groups according to their origin (biological/nonbiological) and fundamental polarimetric properties of retardation, depolarization, and diattenuation. We found that, while biological media are generally depolarizing, retarding, and diattenuating, only one of all the phantoms reviewed incorporated all these properties, and few considered at least combined retardation and depolarization. Samples derived from biological tissue, such as tendon and muscle, remain extremely popular to quickly ascertain a polarimetric system, but do not provide quantifiable results aside from relative direction of their principal optical axis. Microspheres suspensions are the most utilized phantoms for depolarization, and combined with theoretical models can offer true quantification of depolarization or degree of polarization. There is a real paucity of birefringent phantoms despite the retardance being one of the most interesting parameters measurable with polarization techniques. Therefore, future work should be directed at generating truly reliable and repeatable phantoms for this metric determination. Diattenuating phantoms are rare and application-specific. Given that diattenuation is considered to be low in most biological tissues, the lack of such phantoms is seen as less problematic. The heterogeneity of the phantoms reviewed points to a critical need for standardization in this field. Ultimately, all research groups involved in polarimetric studies and instruments development would benefit from sharing a limited set of standardized polarimetric phantoms, as is done earlier in the round robin investigations in ellipsometry.

This guest editorial introduces the special section on Biomedical Imaging and Sensing.

Photoacoustic imaging (PAI) is a fast growing deep-tissue imaging modality. However, light scattering and absorption in biological tissues limit imaging depth. Short near-infrared wavelengths (650 to 950 nm) are widely used for PAI. Using longer near-infrared wavelengths reduces scattering. We demonstrate deep-tissue contrast-enhanced in vivo photoacoustic imaging at a wavelength of 1064 nm. An ultranarrow bandgap semiconducting polymer poly (thienoisoindigo-alt-diketopyrrolopyrrole) (denoted as PIGD) is designed and demonstrated for imaging at 1064 nm. By embedding colloidal nanoparticles (NPs) of PIGD in chicken-breast tissue, an imaging depth of ∼5  cm is achieved. Intravenous injection of PIGD NPs in living rats showed brain vascular images with ∼2 times higher contrast compared with the brain vascular images without any contrast agent. Thus, PIGD NPs as an NIR-II contrast agent opens new opportunities for both preclinical and clinical imaging of deep tissues with enhanced contrast.

To realize visualization of the skin microvascular dysfunction of type 1 diabetic mice, we combined laser speckle contrast imaging and hyperspectral imaging to simultaneously monitor the noradrenaline (NE)-induced responses of vascular blood flow and blood oxygen with the development of diabetes through optical clearing skin window. The main results showed that venous and arterious blood flow decreased without recovery after injection of NE; furthermore, the decrease of arterious blood oxygen induced by NE greatly weakened, especially for 2- and 4-week diabetic mice. This change in vasoconstricting effect of NE was related to the expression of α1-adrenergic receptor. This study demonstrated that skin microvascular function was a potential research biomarker for early warning in the occurrence and development of diabetes. The in vivo skin optical clearing method provides a feasible solution to realize visualization of cutaneous microvessels for monitoring microvascular reactivity under pathological conditions. In addition, visual monitoring of skin microvascular function response has guiding significance for early diagnosis of diabetes and clinical research.

Digital optical phase conjugation (DOPC) enables many optical applications by permitting focusing of light through scattering media. However, DOPC systems require precise alignment of all optical components, particularly of the spatial light modulator (SLM) and camera, in order to accurately record the wavefront and perform playback through the use of time-reversal symmetry. We present a digital compensation technique to optimize the alignment of the SLM in five degrees of freedom, permitting focusing through thick scattering media with a thickness of 5 mm and transport scattering coefficient of 2.5  mm  −  1 while simultaneously improving focal quality, as quantified by the peak-to-background ratio, by several orders of magnitude over an unoptimized alignment.

Point spread function (PSF) engineering has met with lots of interest in various optical imaging techniques, including super-resolution microscopy, microparticle tracking, and extended depth-of-field microscopy. The intensity distributions of the modified PSFs often suffer from deteriorations caused by system aberrations, which greatly degrade the image contrast, resolution, or localization precision. We present an aberration correction method using a spiral-phase-based double-helix PSF as an aberration indicator, which is sensitive and quantitatively correlated to the spherical aberration, coma, and astigmatism. Superior to the routine iteration-based correction methods, the presented approach is iteration-free and the aberration coefficients can be directly calculated with the measured parameters, relieving the computing burden. The validity of the method is verified by both examining the intensity distribution of the conventional Gaussian PSF in three dimensions and observing muntjac skin fibroblast cells. This iteration-free correction method has a potential application in PSF engineering systems equipped with a spatial light modulator.

Skin wrinkling is a typical symptom of cutaneous photoaging; however, the skin wrinkling depends on not only the actual age but also exposure history to ultraviolet B (UVB) rays in individuals. Therefore, there is considerable need for its assessment technique in vivo in skin cosmetics and antiaging dermatology. Wrinkles always appear as linear grooves in the skin, and dermal collagen fibers play an important role to determine the morphology and mechanical properties of the skin. Therefore, an optical probe sensitive to dermal collagen fiber and its orientation will be useful. Polarization-resolved second-harmonic-generation (SHG) microscopy is a promising approach for in vivo evaluation of collagen fiber orientation because the efficiency of SHG light is sensitive to collagen fiber orientation when the incident light is linearly polarized. We investigate orientation change of dermal collagen fiber in prewrinkled and wrinkled skins of the UVB-exposed mouse model using polarization-resolved SHG microscopy. A polarization anisotropic image of the SHG light indicates that the change of collagen fiber orientation starts in the prewrinkled skin of UVB-exposed mice, then the wrinkle appears. Furthermore, the dominant direction of collagen fiber orientation in the prewrinkled skin is significantly parallel to the wrinkle direction in the wrinkled skin. This result implies that the change of collagen fiber orientation is a trigger of wrinkling in cutaneous photoaging.

Mobile phone technology has led to implementation of portable and inexpensive microscopes. Light-emitting diode (LED) array microscopes support various multicontrast imaging by flexible illumination patterns of the LED array that can be achieved without changing the optical components of the microscope. Here, we demonstrate a mobile-phone-based LED array microscope to realize multimodal imaging with bright-field, dark-field, differential phase-contrast, and Rheinberg illuminations using as few as 37 LED bulbs. Using this microscope, we obtained high-contrast images of living cells. Furthermore, by changing the color combinations of Rheinberg illumination, we were able to obtain images of living chromatic structures with enhanced or diminished contrast. This technique is expected to be a foundation for high-contrast microscopy used in modern field studies.

The conical shape of the tympanic membrane (TM or eardrum) plays an important role in its function, such that variations in shape alter the acoustically induced motions of the TM. We present a method that precisely determines both shape and acoustically induced transient response of the entire TM using the same optics and maintaining the same coordinate system, where the TM transient displacements due to a broadband acoustic click excitation (50-μs impulse) and the shape are consecutively measured within <200  ms. Interferograms gathered with continuous high-speed (>2  kHz) optical phase sampling during a single 100-ms wavelength tuning ramp allow precise and rapid reconstructions of the TM shape at varied resolutions (50 to 200  μm). This rapid acquisition of full-field displacements and shape is immune to slow disturbances introduced by breathing or heartbeat of live subjects. Knowledge of TM shape and displacements enables the estimation of surface normal displacements regardless of the orientation of the TM within the measurement system. The proposed method helps better define TM mechanics and provides TM structure and function information useful for the diagnosis of ear disease.

Emerging clinical interest in combining standard white light endoscopy with targeted near-infrared (NIR) fluorescent contrast agents for improved early cancer detection has created demand for multimodal imaging endoscopes. We used two spectrally resolving detector arrays (SRDAs) to realize a bimodal endoscope capable of simultaneous reflectance-based imaging in the visible spectral region and multiplexed fluorescence-based imaging in the NIR. The visible SRDA was composed of 16 spectral bands, with peak wavelengths in the range of 463 to 648 nm and full-width at half-maximum (FWHM) between 9 and 26 nm. The NIR SRDA was composed of 25 spectral bands, with peak wavelengths in the range 659 to 891 nm and FWHM 7 to 15 nm. The spectral endoscope design was based on a “babyscope” model using a commercially available imaging fiber bundle. We developed a spectral transmission model to select optical components and provide reference endmembers for linear spectral unmixing of the recorded image data. The technical characterization of the spectral endoscope is presented, including evaluation of the angular field-of-view, barrel distortion, spatial resolution and spectral fidelity, which showed encouraging performance. An agarose phantom containing oxygenated and deoxygenated blood with three fluorescent dyes was then imaged. After spectral unmixing, the different chemical components of the phantom could be successfully identified via majority decision with high signal-to-background ratio (>3). Imaging performance was further assessed in an ex vivo porcine esophagus model. Our preliminary imaging results demonstrate the capability to simultaneously resolve multiple biological components using a compact spectral endoscopy system.

Pharmacokinetic tomography is emerging as an important methodology for detecting abnormalities in tissue based upon spatially varying estimation of the pharmacokinetic rates governing the leakage of an injected fluorophore between blood plasma and tissue. We present a shape-based reconstruction framework of a compartment-model based formulation of this dynamic fluorescent optical tomography problem to solve for the pharmacokinetic rates and concentrations of the fluorophore from time-varying log intensity measurements of the optical signal. The compartment-model based state variable model is set up in a radial basis function parameterized level set setting. The state (concentrations) and (pharmacokinetic) parameter estimation problem is solved with an iteratively regularized Gauss–Newton filter in a trust-region framework. Reconstructions obtained using this scheme for noisy data obtained from cancer mimicking numerical phantoms of near/sub-cm sizes show a good localization of the affected regions and reasonable estimates of the pharmacokinetic rates and concentration curves.

Surgical excision (Mohs micrographic surgery) is the standard procedure to treat a melanoma, in which an in situ histologic examination of sectioned skin is carried out repeatedly until no cancer cells are detected. The possibility to identify melanoma from the surrounding skin by femtosecond laser-induced breakdown spectroscopy (fs-LIBS) is investigated. For experiments, melanoma induced on a hairless mouse by injection of B16/F10 murine melanoma cell was sampled in the form of frozen tissue sections as in Mohs surgery and analyzed by fs-LIBS (λ  =  1030  nm, τ  =  550  fs). For analysis, the magnesium signal normalized by carbon intensity was utilized to construct an intensity map around the cancer, including both melanoma and surrounding dermis. The intensity map showed a close match to the optically observed morphological and histological features near the cancer region. The results showed that when incorporated into the existing micrographic surgery procedure, fs-LIBS could be a useful tool for histopathologic interpretation of skin cancer possibly with significant reduction of histologic examination time.

Blood coagulation is an important role in the hemostasis process. In the observation using microscopies, an aggregation structure of red blood cells indicates the degree of blood coagulation. Recently, it has been proposed that digital holographic microscopy (DHM) is a powerful tool for biomedical cell imaging on the basis of quantitative phase information. DHM has the advantage in that the real-time and three-dimensional (3-D) quantitative phase imaging can be realized in the wide field of view, which means that the 3-D morphological parameters of biological cells without a staining process are obtained in real time. We report the complete 3-D quantitative phase imaging of blood coagulation structures by optical projection tomography in a flow cytometry using DHM.

Monitoring oxygen saturation (SO₂) in microcirculation is effective for understanding disease dynamics. We have developed an SO₂ estimation method, sidestream dark-field (SDF) oximetry, based on SDF imaging. SDF imaging is a noninvasive and clinically applicable technique to observe microcirculation. We report the first in vivo experiment observing the changes in SO₂ of microcirculation using SDF oximetry. First, heat from the light-emitting diodes used for the SDF imaging might affect hemodynamics in microcirculation, hence, we performed an experiment to evaluate the influence of that on the SDF oximetry. The result suggested that SDF oximetry had enough stability for long-term experiments. Then, to evaluate the sensitivity of SDF oximetry to alterations in the hemodynamics of the microcirculation, we observed the time-lapsed SO₂ changes in the dermis microcirculation of rats under hypoxic stimulation. We confirmed that the SO₂ estimated by SDF oximetry was in accordance with changes in the fraction of inspired oxygen (FiO₂). Thus, SDF oximetry is considered to be able to observe SO₂ changes that occur in accordance with alteration of the microcirculation.

Infrared thermal imaging of brain temperature changes is useful for evaluating cortical activity and disease states, such as stroke. However, the changes depend on a balance between changes in heat generation from metabolism and in heat convection related to blood flow. To discriminate between these effects and gain a clearer understanding of neurovascular metabolic coupling, brain temperature imaging must be improved to measure temperature and blood flow simultaneously. We develop an imaging technique that shows a two-dimensional (2-D) distribution of absolute brain temperature and relative cerebral blood flow changes in anesthetized rats by combining infrared thermal imaging with laser speckle imaging. The changes in brain metabolism and cerebral blood flow are achieved using two different anesthetics (isoflurane and α-chloralose) to evaluate our system. Isoflurane increased cerebral blood flow but decreased metabolism, whereas α-chloralose decreased both parameters. This technique enables simultaneous visualization of brain surface changes in temperature and cerebral blood flow in the same regions. This imaging system will permit further study of neurovascular metabolic coupling in normal and diseased brains.

Photoacoustic (PA) imaging is an emerging imaging technique for many clinical applications. One of the challenges posed by clinical translation is that imaging systems often rely on a finite-aperture transducer rather than a full tomography system. This results in imaging artifacts arising from an underdetermined reconstruction of the initial pressure distribution (IPD). Furthermore, clinical applications often require deep imaging, resulting in a low-signal-to-noise ratio for the acquired signal because of strong light attenuation in tissue. Conventional approaches to reconstruct the IPD, such as back projection and time-reversal, do not adequately suppress the artifacts and noise. We propose a sparsity-based optimization approach that improves the reconstruction of IPD in PA imaging with a linear array ultrasound transducer. In simulation studies, the forward model matrix was measured from k-Wave simulations, and the approach was applied to reconstruct simulated point objects and the Shepp–Logan phantom. The results were compared with the conventional back projection, time-reversal approach, frequency-domain reconstruction, and the iterative least-squares approaches. In experimental studies, the forward model of our imaging system is directly measured by scanning a graphite point source through the imaging field of view. Experimental images of graphite inclusions in tissue-mimicking phantoms are reconstructed and compared with the back projection and iterative least-squares approaches. Overall these results show that our proposed optimization approach can leverage the sparsity of the PA images to improve the reconstruction of the IPD and outperform the existing popular reconstruction approaches.

A clear image of an observed object may deteriorate into unrecognizable speckle when encountering heterogeneous scattering media, thus it is necessary to recover the object image from the speckle. A method combining least square and semidefinite programming is proposed, which can be used for imaging through scattering media. The proposed method consists of two main stages, that is, media scattering characteristics (SCs) estimation and image reconstruction. SCs estimation is accomplished through LS concept after establishing a database of known object-and-speckle pairs. Image reconstruction is realized by solving an SDP problem to obtain the product of the unknown object image and its Hermitian transposition. Finally, the unknown object image can be reconstructed by extracting the largest rank-1 component of the product. Structural similarity (SSIM) index is employed as a performance indicator in speckle prediction and image reconstruction. Numerical simulations and physical experiments are performed to verify the feasibility and practicality of the proposed method. Compared with the existing phase shift interferometry mean square optimization method and the single-shot phase retrieval algorithm, the proposed method is the most precise to obtain the best reconstruction results with highest SSIM index value. The work can be used for exploring the potential applications of scattering media, especially for imaging through turbid media in biomedical, scattering property measurement, and optical image encryption.

An endoscopic optical coherence tomography (OCT) system with a wide field-of-view of 8 mm is presented, which combines the image capability of endoscopic imaging at the middle ear with the advantages of functional OCT imaging, allowing a morphological and functional assessment of the human tympanic membrane. The endoscopic tube has a diameter of 3.5 mm and contains gradient-index optics for simultaneous forward-viewing OCT and video endoscopy. The endoscope allows the three-dimensional visualization of nearly the entire tympanic membrane. In addition, the oscillation of the tympanic membrane is measured spatially resolved and in the frequency range between 500 Hz and 5 kHz with 125 Hz resolution, which is realized by phase-resolved Doppler OCT imaging during acoustical excitation with chirp signals. The applicability of the OCT system is demonstrated in vivo. Due to the fast image acquisition, structural and functional measurements are only slightly affected by motion artifacts.

The reflection and transmission of coherent light from a biological system can yield information about its condition. In the case of blood exposed to the air, there is a change in the properties of the speckle patterns observed in the coagulation process. This can be studied by means of the rate of temporal variation, the contrast, and also the fractality of patterns. The fractality of the speckle pattern can be investigated by a fractal dimension, which can quantify a level of the complexity of platelet aggregation structure and a fibrin network formed in the process of blood coagulation. In addition, dynamic characteristics of a movement in blood also contain information on the progress of the coagulation process. Fractality and dynamic characteristics are investigated simultaneously for speckle patterns observed in the coagulation process of stored horse blood. Experimental results show the feasibility of the proposed method for detecting hemolysis and formation of platelet aggregation structure and the fibrin network during the coagulation process.

The aim of this study is to evaluate the osteoblastic collagen synthesis under mechanical stimulation using second-harmonic-generation (SHG) microscopy. We apply SHG microscopy to monitor the collagen fibers synthesized by osteoblast-like cells (MC3T3-E1) without the need for fixation and staining. To quantitatively evaluate the influence of mechanical stimulation on osteoblastic collagen synthesis, we compare SHG images of osteoblast-synthesized collagen fibers with and without a cyclic stretch stimulus applied using a lab-made stretching device. We acquire SHG images every 7 days for 3 weeks at different stimulus conditions (5 min/day and 3 h/day with a strain magnitude of 5% and a frequency of 0.5 Hz). Image analysis of the average SHG intensity indicates that the amount of osteoblastic collagen synthesis is significantly enhanced by the cyclic stretch compared with the nonstretched condition, while there is no significant difference between the two mechanical stimulation conditions. Furthermore, the maturity of the collagen fibers in the early stage of bone formation is not affected by the mechanical stimulation. The results can be used in bone regenerative medicine to apply feedback control of collagen synthesis by artificial stimulation.

We investigate a multispectral imaging method to evaluate spatiotemporal changes in both cutaneous hemoglobin concentration and light scattering parameter in mouse skin through diffuse reflectance spectroscopy using the reflectance images acquired at isosbestic wavelengths of hemoglobin (420, 450, 500, and 585 nm). In the proposed approach, Monte Carlo simulation-based empirical formulas are introduced to extract the scattering power b representing the wavelength dependence of light scattering spectrum of skin tissue, as well as the total hemoglobin concentration C_th in dermal vasculatures. The use of isosbestic wavelengths of hemoglobin enables the values of C_th and b to be estimated independently of the oxygenation of hemoglobin. Experiments using in vivo mice two-stage chemical carcinogenesis model are performed to confirm the feasibility of the proposed method for evaluating the changes in cutaneous vasculatures and tissue morphology during tumor initiation, promotion, and progression processes. The experimental results reveal that the changes in scattering power b of back skin are significantly reduced and followed by the increase in total hemoglobin concentration C_th in the carcinogenesis mice group, which indicates morphological changes in skin tissue such as edema and cell swelling caused by tumor promotion and successive angiogenesis along with tumor progression. The results suggest that the potential of the present method to detect cutaneous carcinogenesis in an early stage and monitor physiological changes during promotion and progression process of nonmelanoma tumors.

The generalized phase contrast method is employed as an efficient “phase-only” laser beam-shaping technique in an optical setup built for catapulting microspheres through simple mucus models. The influence of the laser power and mucin concentration on the motion of the microspheres is investigated in terms of instant and average velocities on a 250-μm trajectory, corresponding to the mucus thickness in the human gastrointestinal tract. Increasing the laser power leads to higher velocities in all the tested samples, while increasing the mucin concentration leads to significant velocity decrease for similar laser input power. However, velocities of up to 95  μm  ·  s^−  1 are demonstrated in a 5% mucin simple mucus model using our catapulting system. This study contributes to understanding and overcoming the challenges of optical manipulation in mucus models. This paves the way for efficient optical manipulation of three-dimensional-printed light-controlled microtools with the ability to penetrate the mucus biobarrier for in vitro drug-delivery studies.

The optical attenuation coefficient (OAC) reflects the optical properties of various tissues or tissues of the same type under different physiological conditions. Quantitative measurement of OAC from optical coherence tomography (OCT) signals can provide additional information and can increase the potential for OCT applications. We present an optimized depth-resolved estimation (ODRE) method that derives a precise mapping between the measured OCT signal and the OAC. In contrast to previous depth-resolved estimation (DRE) methods, the optimized method can estimate the OAC in any depth range and ignore whether the light is completely attenuated. Numerical simulations and phantom experiments are used to verify its validity, and this method is applied to detect cerebral damage. In combination with OCT angiography, real-time observation of the change of blood perfusion and the degree of cerebral damage in mice with focal cerebral ischemia provides important information to help us understand the temporal relationship between brain damage and ischemia.

The refractive index of blood is a key biophysical parameter, which can reflect the physiological state. We measured the refractive index of whole blood and other components, such as serum, plasma, and hemoglobin, based on internal reflection by using a homemade apparatus in the spectral range of 400 to 750 nm. In addition to the hemoglobin solution, which has a Soret band about 420 nm and two Q-bands between 500 and 600 nm, the measurements of other samples are the normal dispersion curve. The results are approximated by the Cauchy equation and Sellmeier equation, and the correlation coefficients are more than 0.997.

The observed behavior of short-wave infrared (SWIR) light in tissue, characterized by relatively low scatter and subdiffuse photon transport, has generated considerable interest for the potential of SWIR imaging to produce high-resolution, subsurface images of fluorescence activity in vivo. These properties have important implications for fluorescence-guided surgery and preclinical biomedical research. Until recently, translational efforts have been impeded by the conventional understanding that fluorescence molecular imaging in the SWIR regime requires custom molecular probes that do not yet have proven safety profiles in humans. However, recent studies have shown that two readily available near-infrared (NIR-I) fluorophores produce measurable SWIR fluorescence, implying that other conventional fluorophores produce detectable fluorescence in the SWIR window. Using SWIR spectroscopy and wide-field SWIR imaging with tissue-simulating phantoms, we characterize and compare the SWIR emission properties of eight commercially available red/NIR-I fluorophores commonly used in preclinical and clinical research, in addition to a SWIR-specific fluorophore. All fluorophores produce measurable fluorescence emission in the SWIR, including shorter wavelength dyes such as Alexa Fluor 633 and methylene blue. This study is the first to report SWIR fluorescence from six of the eight conventional fluorophores and establishes an important comparative reference for developing and evaluating SWIR imaging strategies for biomedical applications.

Blast-induced traumatic brain injury has been a recent major concern in neurotraumatology. However, its pathophysiology and mechanism are not understood partly due to insufficient information on the brain pathophysiology during/immediately after shock wave exposure. We transcranially applied a laser-induced shock wave (LISW, ∼19  Pa  ·  s) to the left frontal region in a rat and performed multispectral imaging of the ipsilateral cortex through a cranial window (n  =  4). For the spectral data obtained, we conducted multiple regression analysis aided by Monte Carlo simulation to evaluate vascular diameters, regional hemoglobin concentration (rC_Hb), tissue oxygen saturation (StO₂), oxygen extraction fraction, and light-scattering signals as a signature of cortical spreading depolarization (CSD). Immediately after LISW exposure, rC_Hb and StO₂ were significantly decreased with distinct venular constriction. CSD was then generated and was accompanied by distinct hyperemia/hyperoxemia. This was followed by oligemia with arteriolar constriction, but it soon recovered (within ∼20  min). However, severe hypoxemia was persistently observed during the post-CSD period (∼1  h). These observations indicate that inadequate oxygen supply and/or excessive oxygen consumption continued even after blood supply was restored in the cortex. Such a hypoxemic state and/or a hypermetabolic state might be associated with brain damage caused by a shock wave.

We compare previously reported benzoporphyrin derivative (BPD)-mediated photodynamic therapy (PDT) results for reactive singlet oxygen concentration (also called singlet oxygen dose) on mice with simulations using a computational device, Dosie™, that calculates light transport and photokinetics for PDT in near real-time. The two sets of results are consistent and validate the use of the device in PDT treatment planning to predict BPD-mediated PDT outcomes in mice animal studies based on singlet oxygen dose, which showed a much better correlation with the cure index than the conventional light dose.

Two-thirds of out-of-hospital cardiac arrest patients, who survive to hospital admission, die in the hospital from neurological injuries related to cerebral hypoperfusion. Therefore, noninvasive real-time monitoring of the cerebral oxygen metabolism in cardiac arrest patients is extremely important. Hyperspectral near-infrared spectroscopy (hNIRS) is a noninvasive technique that measures concentrations of the key chromophores in the brain, such as oxygenated hemoglobin, deoxygenated hemoglobin, and cytochrome C oxidase (CCO), an intracellular marker of oxygen consumption. We tested hNIRS on 10 patients undergoing transcatheter aortic valve insertion, where rapid ventricular pacing (RVP) is required to temporarily induce sudden hypotension and hypoperfusion that mimic cardiac arrest. Using multidistance hNIRS, we found that tissue oxygen saturation changes in the cerebral tissue were lower than those in the scalp during RVP. CCO redox changes were detected in cerebral tissue but not in the scalp during RVP. We have demonstrated that hNIRS is feasible and can detect sudden changes in cerebral oxygenation and metabolism in patients during profound hypotension.

We report the ex vivo results of an in-house fabricated portable device based on polarized fluorescence measurements in the clinical environment. This device measures the polarized fluorescence and elastic scattering spectra with 405-nm laser and white light sources, respectively. The dominating fluorophore with 405-nm excitation is flavin adenine dinucleotide (FAD) with a fluorescence peak around 510 nm. The measured spectra are highly modulated by the interplay of scattering and absorption effects. Due to this, valuable information gets masked. To reduce these effects, intrinsic fluorescence was extracted by normalizing polarized fluorescence spectra with polarized elastic scattering spectra obtained. A number of fluorophores contribute to the fluorescence spectra and need to be decoupled to understand their roles in the progression of cancer. Nelder–Mead method has been utilized to fit the spectral profile with Gaussian to decouple the different bands of contributing fluorophores (FAD and porphyrin). The change in concentration of FAD during disease progression manifests in the change in ratio of total area to FWHM of its Gaussian profile. Receiver operating characteristic (ROC) curve analysis has been used to discriminate different grades of cervical precancer by using the ratio as input parameter. The sensitivity and specificity for discrimination of normal samples from CIN I (cervical intraepithelial neoplasia) are 75% and 54%, respectively. Further, the normal samples can be discriminated from CIN II samples with 100% and 82% sensitivity and specificity, respectively, and the CIN I from CIN II samples can also be discriminated with 100% sensitivity and 90% specificity, respectively. The results show that the change in the concentration of (FAD) can be used as a marker to discriminate the different grades of the cancer and biochemical changes at an early stage of the cancer can also be monitored with this technique.

Cherenkov emission generated in tissue during radiotherapy can be harnessed for the imaging biochemistry of tissue microenvironments. Cherenkov-excited luminescence scanned imaging (CELSI) provides a way to optically and noninvasively map oxygen-related signals, which is known to correlate to outcomes in radiotherapy. Four candidate phosphorescent reagents PtG4, MM2, Ir(btb)₂  (  acac  )  , and MitoID were studied for oxygen sensing, testing in a progressive series of (a) in solution, (b) in vitro, and (c) in subcutaneous tumors. In each test, the signal strength and response to oxygen were assessed by phosphorescence intensity and decay lifetime measurement. MM2 showed the most robust response to oxygen changes in solution, followed by PtG4, Ir(btb)₂  (  acac  )  , and MitoID. However, in PANC-1 cells, their oxygen responses differed with Ir(btb)₂  (  acac  )   exhibiting the largest phosphorescent intensity change in response to changes in oxygenation, followed by PtG4, MM2, and MitoID. In vivo, it was only possible to utilize Ir(btb)₂  (  acac  )   and PtG4, with each being used at nanomole levels, to determine signal strength, lifetime, and pO₂. Oxygen sensing with CELSI during radiotherapy is feasible and can estimate values from 1 mm regions of tissue when used in the configuration of this study. PtG4 was the most amenable to in vivo sensing on the timescale of external beam LINAC x-rays.

Most new caries lesions are found in the pits and fissures of the occlusal surface. Radiographs have extremely low sensitivity for early occlusal decay, and by the time the lesion is severe enough to appear on a radiograph, it typically has penetrated well into the dentin and surgical intervention is required. The occlusal surfaces are often heavily stained, and visual and tactile detection have poor sensitivity and specificity. Previous near-infrared imaging studies at wavelengths beyond 1300 nm have demonstrated that stains are not visible and demineralization on the occlusal surfaces can be viewed without interference from stains. The objective of our study is to determine how the contrast between sound and lesion areas on occlusal surfaces varies with wavelength from the visible to 2350 nm and determine to what degree stains interfere with that contrast. The lesion contrast for reflectance is measured in 55 extracted teeth with suspected occlusal lesions from 400 to 2350 nm employing silicon and indium gallium arsenide imaging arrays. In addition, the lesion contrast is measured on 25 extracted teeth with suspected occlusal lesions from 400 to 1600 nm in reflectance and from 830 to 1400 nm in transillumination before and after stains are removed using a ultrasonic scaler. The highest lesion contrast in reflectance is measured at wavelengths >1700  nm. Stains interfere significantly at wavelengths <1150  nm (400 to 1150) for both reflectance and transillumination measurements. Our study suggests that the optimum wavelengths for imaging decay in the occlusal surfaces are >1700  nm for reflectance (1700 to 2350 nm) and near 1300 nm (1250 to 1350 nm) for transillumination.

When monitoring a moist sample using mid-infrared spectroscopy, its thickness must be <100  μm to avoid light absorption from the water. Therefore, we propose an ultrasonic-assisted mid-infrared spectroscopic imaging method that can generate a reflection plane at a depth of 100  μm from the surface of the sample by creating an ultrasonic standing wave. A frequency of 10 MHz is required to obtain an optical path length of 100  μm in biological samples. However, because biological samples generally have high compressibility, attenuation of ultrasonic waves at this frequency is significant. We use agar as a biological phantom and observe that a reflection plane is generated inside by ultrasonic standing waves using optical coherence tomography. It is found that when the sample is vibrated with an 800-kHz ultrasonic wave, a reflection plane is generated at a depth shallower than the theoretically predicted value. We believe that the reflection plane is generated by parametric standing waves, which are based on parametric effect. We detect the waveform distortion using an acoustic emission sensor and confirm the higher harmonics that generate the observed reflection plane using a fast Fourier transform.

Cataract is one of the most prevalent causes of blindness around the world. Understanding the mechanisms of cataract development and progression is important for clinical diagnosis and treatment. Cold cataract has proven to be a robust model for cataract formation that can be easily controlled in the laboratory. There is evidence that the biomechanical properties of the lens can be significantly changed by cataract. Therefore, early detection of cataract, as well as evaluation of therapies, could be guided by characterization of lenticular biomechanical properties. In this work, we utilized optical coherence elastography (OCE) to monitor the changes in biomechanical properties of ex vivo porcine lenses during formation of cold cataract. Elastic waves were induced in the porcine lenses by a focused micro air-pulse while the lenses were cooled, and the elastic wave velocity was translated to Young’s modulus of the lens. The results show an increase in the stiffness of the lens due to formation of the cold cataract (from 11.3  ±  3.4 to 21.8  ±  7.8  kPa). These results show a relation between lens opacity and stiffness and demonstrate that OCE can assess lenticular biomechanical properties and may be useful for detecting and potentially characterizing cataracts.

Optical coherence tomography angiography (OCTA) has been widely used for en face visualization of the microvasculature, but is challenged for real three-dimensional (3-D) topologic imaging due to the “tail” artifacts that appear below large vessels. Further, OCTA is generally incapable of differentiating descending arterioles from ascending venules. We introduce a normalized field autocorrelation function-based OCTA (g₁-OCTA), which minimizes the tail artifacts and is capable of distinguishing penetrating arterioles from venules in the 3-D image. g₁   (  τ  )   is calculated from repeated optical coherence tomography (OCT) acquisitions for each spatial location. The decay amplitude of g₁   (  τ  )   is retrieved to represent the dynamics for each voxel. To account for the small g₁   (  τ  )   decay in capillaries where red blood cells are flowing slowly and discontinuously, Intralipid is injected to enhance the OCT signal. We demonstrate that the proposed technique realizes 3-D OCTA with negligible tail projections and the penetrating arteries are readily identified. In addition, compared to regular OCTA, the proposed g₁-OCTA largely increased the depth-of-field. This technique provides a more accurate rendering of the vascular 3-D anatomy and has the potential for more quantitative characterization of vascular networks.

A segmentation method based on phase retardation measurements from polarization-sensitive optical coherence tomography (PS-OCT) is developed to differentiate the structural zones of articular cartilage. The organization of collagen matrix in articular cartilage varies over the different structural zones, generating different tissue birefringence. Analyzing the slope of the accumulated phase retardation at different depths can detect the variation in tissue birefringence and be used to segment the structural zones. The method is validated on phantoms composed of layers of different materials. Articular cartilage samples from adult swine are segmented with the method. The characteristics in each segmented zone are also examined by histology and high-resolution second-harmonic generation imaging, showing distinctive properties that match with the anatomical structure of articular cartilage. The segmentation algorithm is also applied on PS-OCT images acquired at multiple illumination angles, where the angular dependence of tissue birefringence in the deep zone is detected. This method offers a noninvasive imaging approach to differentiating the structural zones of articular cartilage, as well as a quantification approach based on the phase retardation measurements of PS-OCT. This method has great potential in studying depth-related progression of cartilage degeneration.

For patients undergoing surgical cancer resection of squamous cell carcinoma (SCCa), cancer-free surgical margins are essential for good prognosis. We developed a method to use hyperspectral imaging (HSI), a noncontact optical imaging modality, and convolutional neural networks (CNNs) to perform an optical biopsy of ex-vivo, surgical gross-tissue specimens, collected from 21 patients undergoing surgical cancer resection. Using a cross-validation paradigm with data from different patients, the CNN can distinguish SCCa from normal aerodigestive tract tissues with an area under the receiver operator curve (AUC) of 0.82. Additionally, normal tissue from the upper aerodigestive tract can be subclassified into squamous epithelium, muscle, and gland with an average AUC of 0.94. After separately training on thyroid tissue, the CNN can differentiate between thyroid carcinoma and normal thyroid with an AUC of 0.95, 92% accuracy, 92% sensitivity, and 92% specificity. Moreover, the CNN can discriminate medullary thyroid carcinoma from benign multinodular goiter (MNG) with an AUC of 0.93. Classical-type papillary thyroid carcinoma is differentiated from MNG with an AUC of 0.91. Our preliminary results demonstrate that an HSI-based optical biopsy method using CNNs can provide multicategory diagnostic information for normal and cancerous head-and-neck tissue, and more patient data are needed to fully investigate the potential and reliability of the proposed technique.

Phase and fluorescence are complementary contrasts that are commonly used in biology. However, the coupling of these two modalities is traditionally limited to high magnification and complex imaging systems. For statistical studies of biological populations, a large field-of-view is required. We describe a 30  mm² field-of-view dual-modality mesoscope with a 4-μm resolution. The potential of the system to address biological questions is illustrated on white blood cell numeration in whole blood and multiwavelength imaging of the human osteosarcoma (U2-OS) cells.

We report a fiber-optic plasmonic probe with nanogap-rich gold nanoislands for on-site surface-enhanced Raman spectroscopy (SERS). The plasmonic probe features nanogap-rich Au nanoislands on the top surface of a single multimode fiber. Au nanoislands were monolithically fabricated using repeated solid-state dewetting of thermally evaporated Au thin film. The plasmonic probe shows 7.8  ×  10⁶ in SERS enhancement factor and 100 nM in limit-of-detection for crystal violet under both the excitation of laser light and the collection of SERS signals through the optical fiber. The fiber-through measurement also demonstrates the label-free SERS detection of folic acid at micromolar level. The plasmonic probe can provide a tool for on-site and in vivo SERS applications.

Our preliminary study investigates an automated, vibrating fiber optic tip for dusting of kidney stones during thulium fiber laser (TFL) lithotripsy. A (0.75-mm diameter and 5-mm length) magnetic bead was attached to the fiber jacket, centered 2 cm from distal fiber tip. A solenoid was placed parallel to the fiber with a 0.5-mm gap between solenoid and magnetic bead on fiber. The solenoid was used to create a magnetic force on the bead, inducing fiber vibration. Calibration tests for fiber motion in both air and water were performed. The ablation crater characteristics (surface area, volume, depth, and major/minor axis) of uric acid stones were measured using optical coherence tomography, after delivery of 1500 TFL pulses at 1908 nm, 33 mJ, 500  μs, and up to 300 Hz, through 50-, 100-, and 150-μm-core fibers. The resonant frequency was dependent on fiber diameter and rigidity, with a cutoff pivot point for optimum vibration amplitude at 4 cm. Maximum fiber displacement is about 1 mm in water and 4 mm in air. For 50-, 100-, and 150-μm-core fibers, ablated surface area averaged 1.7, 1.7, and 2.8 times greater with vibrating fiber than fixed fiber, respectively. For these fibers, ablation volume averaged 1.1, 1.5, and 1.1 times greater with vibrating fiber than fixed fiber, given a fixed energy per pulse, respectively. Our preliminary study demonstrates the functionality of an automated, vibrating fiber system for stone “dusting,” with significantly larger surface area but similar ablation volumes as a fixed fiber. Future studies will focus on optimization of fiber parameters (especially displacement) and miniaturization of system components to facilitate integration into ureteroscopes.