Photobiomodulation (PBM) therapy, previously known as low-level laser therapy, was discovered more than 50 years ago, yet there is still no agreement on the parameters and protocols for its clinical application. Some groups have recommended the use of a power density less than 100  mW/cm² and an energy density of 4 to 10  J/cm² at the level of the target tissue. Others recommend as much as 50  J/cm² at the tissue surface. The wide range of parameters that can be applied (wavelength, energy, fluence, power, irradiance, pulse mode, treatment duration, and repetition) in some cases has led to contradictory results. In our review, we attempt to evaluate the range of effective and ineffective parameters in PBM. Studies in vitro with cultured cells or in vivo with different tissues were divided into those with higher numbers of mitochondria (muscle, brain, heart, nerve) or lower numbers of mitochondria (skin, tendon, cartilage). Graphs were plotted of energy density against power density. Although the results showed a high degree of variability, cells/tissues with high numbers of mitochondria tended to respond to lower doses of light than those with lower number of mitochondria. Ineffective studies in cells with high mitochondrial activity appeared to be more often due to over-dosing than to under-dosing.

This guest editorial introduces the special section honoring Prof. Steven L. Jacques.

We hypothesize that the capsular optical properties and thickness combined affect how accurate the diffuse reflectance on the surface of a capsular solid organ represents that on the subcapsular parenchyma. Monte Carlo simulations on two-layer geometries evaluated how a thin superficial layer with the thickness from 10 to 1000  μm affected the surface diffuse reflectance over a source–detector separation spanning 0.01 to 10 mm. The simulations represented the superficial layer presenting various contrasts concerning refractive index, anisotropy factor, absorption coefficient, and reduced scattering coefficient, versus those of the subsurface main medium. An analytical approach modeled the effects of the superficial layer of various thicknesses and optical properties on diffuse reflectance. Diffuse reflectance spectroscopy was performed ex vivo on 10 fresh human livers and 9 fresh human kidneys using a surface probe with a 3-mm source–detector separation. The difference of the device-specific diffuse reflectance on the organ between with the capsule and without the capsule has significantly greater spectral variation in the kidney than in the liver. The significantly greater spectral deviation of surface diffuse reflectance between with and without the capsule in the kidney than in the liver was analytically accountable by considering the much thicker capsule of the kidney than of the liver.

Microscopy with ultraviolet surface excitation (MUSE) is investigated as a means to enhance curricula and education in the life sciences based on simplicity of use, the incorporation of inexpensive hardware, and the simplest methods of tissue preparation. Ultraviolet excitation in effect replaces tissue sectioning because it penetrates only a few micrometers below the tissue surface at the single cell level, preventing the generation of out-of-focus light. Although tissue autofluorescence may be used, image quality and content can be enhanced by a brief immersion in a solution of nontoxic fluorescent dyes that selectively highlight different cellular compartments. Safe mixed-dye powder combinations have been developed to provide students who have minimal lab proficiencies with a one-step tissue staining process for rapid tissue preparation.

In linear-array transducer-based photoacoustic (PA) imaging, B-scan PA images are formed using the raw channel PA signals. Delay-and-sum (DAS) is the most prevalent algorithm due to its simple implementation, but it leads to low-quality images. Delay-multiply-and-sum (DMAS) provides a higher image quality in comparison with DAS while it imposes a computational burden of O  (  M²  )  . We introduce a nonlinear (NL) beamformer for linear-array PA imaging, which uses the p’th root of the detected signals and imposes the complexity of DAS [O  (  M  )  ]. The proposed algorithm is evaluated numerically and experimentally [wire-target and in-vivo sentinel lymph node (SLN) imaging], and the effects of the parameter p are investigated. The results show that the NL algorithm, using a root of p (NL_p), leads to lower sidelobes and higher signal-to-noise ratio compared with DAS and DMAS, for (p  >  2). The sidelobes level (for the wire-target phantom), at the depth of 11.4 mm, are about −31, −52, −52, −67, −88, and −109  dB, for DAS, DMAS, NL_2, NL_3, NL_4, and NL_5, respectively, indicating the superiority of the NL_p algorithm. In addition, the best value of p for SLN imaging is reported to be 12.

Annually, about 15 million preterm infants are born in the world. Of these, due to complications resulting from their premature birth, about 1 million would die before the age of five. Since the high incidence of preterm birth (PTB) is partially due to the lack of effective diagnostic modalities, methodologies are needed to determine risk of PTB. We propose a noninvasive tool based on polarized light imaging aimed at measuring the organization of collagen in the cervix. Cervical collagen has been shown to remodel with the approach of parturition. We used a full-field Mueller matrix polarimetric colposcope to assess and compare cervical collagen content and structure in nonpregnant and pregnant women in vivo. Local collagen directional azimuth was used and a total of eight cervices were imaged.

The optimal photoacoustic probe design is the key to obtain highest imaging sensitivity in photoacoustic computed tomography. Two commonly used probe design types are dark- and bright-field designs. We proposed a design for photoacoustic probe called quasibright-field illumination and compared the performance of all three kinds of probes theoretically and experimentally. Our conclusion is that the proposed quasibright-field illumination photoacoustic probe is superior compared to the existing probe designs as demonstrated. However, each type of illumination should still have its own advantages under certain circumstances. The dark-field illumination is capable of minimizing surface interference signals and reducing their contributions to the background of deeper signals. Hence, it should perform better when imaging samples with high optical absorbance at the surface layer. The bright field may perform better under circumstance when phase distortion is less. We also designed and fabricated three kinds of probes using a single multimode optical fiber for laser energy delivery instead of fiber bundle. Single fiber probes are low cost, transmit laser energy efficiently, and are compact for easy handling. Thus, our study not only provides a method for probe design but also a guidance for cost-effective transducer array-based photoacoustic probe design and manufacturing in the future.

Photoacoustic imaging using a focused, scanning detector in combination with a pulsed light source is a common technique to visualize light-absorbing structures in biological tissue. In the acoustic resolution mode, where the imaging resolution is given by the properties of the transducer, there are various challenges related to the choice of sensors and the optimization of the illumination. These are addressed by linking a Monte Carlo simulation of energy deposition to a time-domain model of acoustic propagation and detection. In this model, the spatial and electrical impulse responses of the focused transducer are combined with a model of acoustic attenuation in a single response matrix, which is used to calculate detector signals from a volumetric distribution of absorbed energy density. Using the radial symmetry of the detector, the calculation yields a single signal in less than a second on a standard personal computer. Various simulation results are shown, comparing different illumination geometries and demonstrating spectral imaging. Finally, simulation results and experimental images of an optically characterized phantom are compared, validating the accuracy of the model. The proposed method will facilitate the design of photoacoustic imaging devices and will be used as an accurate forward model for iterative reconstruction techniques.

We have determined in vivo optical scattering properties of normal human skin in 1734 subjects, mostly with fair skin type, within the Swedish CArdioPulmonary bioImage Study. The measurements were performed with a noninvasive system, integrating spatially resolved diffuse reflectance spectroscopy and laser Doppler flowmetry. Data were analyzed with an inverse Monte Carlo algorithm, accounting for both scattering, geometrical, and absorbing properties of the tissue. The reduced scattering coefficient was found to decrease from 3.16 ± 0.72 to 1.13 ± 0.27 mm−1 (mean ± SD) in the 475- to 850-nm wavelength range. There was a negative correlation between the reduced scattering coefficient and age, and a significant difference between men and women in the reduced scattering coefficient as well as in the fraction of small scattering particles. This large study on tissue scattering with mean values and normal variation can serve as a reference when designing diagnostic techniques or when evaluating the effect of therapeutic optical systems.

Absorption effect is a basic optical phenomenon and an important feature in tissue imaging and characterization. Based on our Monte Carlo simulation on the anisotropic tissue model (sphere-cylinder birefringence model), combined with our experiments of tissue phantoms, we demonstrate the influence of absorption effect on Mueller matrix and particularly on depolarization, linear retardance, and diattenuation parameters. The simulation and experimental results show a good consistency on the suppressed depolarization and scatterering induced retardance, and the enhanced diattenuation caused by the absorption, and also indicate the birefringence induced retardance insensitive to the absorption. Study of the phase function of different incident polarized lights and the distribution of scattering number gives a preliminary explanation about the above results.

X-ray and optical technologies are the two central pillars for human imaging and therapy. The strengths of x-rays are deep tissue penetration, effective cytotoxicity, and the ability to image with robust projection and computed-tomography methods. The major limitations of x-ray use are the lack of molecular specificity and the carcinogenic risk. In comparison, optical interactions with tissue are strongly scatter dominated, leading to limited tissue penetration, making imaging and therapy largely restricted to superficial or endoscopically directed tissues. However, optical photon energies are comparable with molecular energy levels, thereby providing the strength of intrinsic molecular specificity. Additionally, optical technologies are highly advanced and diversified, being ubiquitously used throughout medicine as the single largest technology sector. Both have dominant spatial localization value, achieved with optical surface scanning or x-ray internal visualization, where one often is used with the other. Therapeutic delivery can also be enhanced by their synergy, where radio-optical and optical-radio interactions can inform about dose or amplify the clinical therapeutic value. An emerging trend is the integration of nanoparticles to serve as molecular intermediates or energy transducers for imaging and therapy, requiring careful design for the interaction either by scintillation or Cherenkov light, and the nanoscale design is impacted by the choices of optical interaction mechanism. The enhancement of optical molecular sensing or sensitization of tissue using x-rays as the energy source is an important emerging field combining x-ray tissue penetration in radiation oncology with the molecular specificity and packaging of optical probes or molecular localization. The ways in which x-rays can enable optical procedures, or optics can enable x-ray procedures, provide a range of new opportunities in both diagnostic and therapeutic medicine. Taken together, these two technologies form the basis for the vast majority of diagnostics and therapeutics in use in clinical medicine.

This ex-vivo study evaluates the feasibility of diffuse reflectance spectroscopy (DRS) for discriminating tumor from healthy tissue, with the aim to develop a technology that can assess resection margins for the presence of tumor cells during oral cavity cancer surgery. Diffuse reflectance spectra were acquired on fresh surgical specimens from 28 patients with oral cavity squamous cell carcinoma. The spectra (400 to 1600 nm) were detected after illuminating tissue with a source fiber at 0.3-, 0.7-, 1.0-, and 2.0-mm distances from a detection fiber, obtaining spectral information from different sampling depths. The spectra were correlated with histopathology. A total of 76 spectra were obtained from tumor tissue and 110 spectra from healthy muscle tissue. The first- and second-order derivatives of the spectra were calculated and a classification algorithm was developed using fivefold cross validation with a linear support vector machine. The best results were obtained by the reflectance measured with a 1-mm source–detector distance (sensitivity, specificity, and accuracy are 89%, 82%, and 86%, respectively). DRS can accurately discriminate tumor from healthy tissue in an ex-vivo setting using a 1-mm source–detector distance. Accurate validation methods are warranted for larger sampling depths to allow for guidance during oral cavity cancer excision.

Optical spectral images can be used to estimate the amount of bulk absorbers in tissues, specifically oxy- and deoxyhemoglobin, as well as scattering parameters. Most systems that capture spectral image data are large, heavy, and expensive. This paper presents a full end-to-end analysis of a low-cost reflectance-mode multispectral imaging system operating in the visible and near-infrared spectra. The system consists of 13 LEDs mounted on a printed circuit board, a monochrome machine vision camera, and a tablet computer to control the hardware. The bill of materials for the system is less than $1000. Hardware design and implementation are detailed. Calibration, image capture, and preprocessing are also discussed. In validation experiments, excellent agreement is observed in diffuse reflectance measurements between the spectral camera setup and a spectrometer. To demonstrate that such spectral image data can yield meaningful optical measurements in vivo, the forearms of eight volunteers are imaged in the system. Their data are then analyzed to estimate the tissue optical properties of different skin layers using a Monte Carlo lookup table. In three volunteers, spectral images are captured before and after inducing erythema using a warm wet towel. Across the three subjects, a clear increase in the blood content of the superficial plexus layer was observed as a result of the erythema. Collectively, these findings suggest that a low-cost system can capture accurate spectral data and that clinically meaningful information can be derived from it.

Raman spectroscopy is widely used to investigate the structure and property of the molecules from their vibrational transitions and may allow for the diagnosis of cancer in a fast, objective, and nondestructive manner. This experimental study aims to propose the use of the 1064-nm wavelength laser in a Raman spectroscopy and to evaluate its discrimination capability in prostate cancer diagnosis. Seventy-four spectra from patients who underwent radical prostatectomy were evaluated. The acquired signals were filtered, normalized, and corrected for possible oscillations in the laser intensity and fluorescence effects. Wilcoxon tests revealed significant differences between the benign and malign samples associated with the deformation vibration characteristic of nucleic acids, proteins, and lipids. A classifier based on support vector machines was able to predict the Gleason scores of the samples with 95% of accuracy, opening a perspective for the use of the 1064-nm excitatory wavelength in prostatic cancer diagnosis.

The tympanic membrane (TM) separates the outer ear from the tympanic cavity. Repeated pathologies can permanently decrease its tension, inducing conductive hearing loss and adhesive processes up to cholesteatoma. The current main therapy is its surgical reconstruction. Even though lasers have been proposed to tighten atrophic TMs, details of this effect, specifically histological analyses, are missing. We therefore used laser pulses to induce TM collagen remodeling in an animal model to compare the histological and electrophysiological effects of different applied laser intensities before entering clinical studies. We irradiated Fuchsin-stained areas of the TM in anesthetized mice with 532-nm laser-pulses of 10 mW for 30 s (0.3 J), 25 mW for 30 s (0.75 J) or 50 mW for 30 s (1.5 J) monitoring hearing with auditory brainstem responses (ABRs). The mice were sacrificed after 2 to 8 weeks and histologically analyzed. An increase in the TM thickness within the defined, stained, and irradiated areas could be observed after 4 weeks. Polarized light microscopy and transmission electron microscopy demonstrated the tissue volume increase majorly due to new collagen-fibrils. Directly after irradiation, ABR thresholds did not increase. We herein demonstrate a controlled laser-induced collagen remodeling within defined areas of the TM. This method might be the prophylactic solution for chronic inflammatory ear pathologies related to decreased TM tension.

We report a near-infrared spectroscopy (NIRS) study of coherent hemodynamic oscillations measured on the human forehead at multiple source–detector distances (1 to 4 cm). The physiological source of the coherent hemodynamics is arterial blood pressure oscillations at a frequency of 0.1 Hz, induced by cyclic inflation (to a pressure of 200 mmHg) and deflation of two thigh cuffs wrapped around the subject’s thighs. To interpret our results, we use a recently developed hemodynamic model and a phasor representation of the oscillations of oxyhemoglobin, deoxyhemoglobin, and total hemoglobin concentrations in the tissue (phasors O, D, and T, respectively). The increase in the phase angle between D and O at larger source–detector separations is assigned to greater flow versus volume contributions and to a stronger blood flow autoregulation in deeper tissue (brain cortex) with respect to superficial tissue (scalp and skull). The relatively constant phase lag of T versus arterial blood pressure oscillations at all source–detector distances was assigned to competing effects from stronger autoregulation and smaller arterial-to-venous contributions in deeper tissue with respect to superficial tissue. We demonstrate the application of a hemodynamic model to interpret coherent hemodynamics measured with NIRS and to assess the different nature of shallow (extracerebral) versus deep (cerebral) tissue hemodynamics.

Pulsed dye laser irradiation in the wavelength range of 585 to 600 nm is currently the gold standard for treatment of port-wine stains (PWSs). However, this treatment method is often ineffective for deeply seated blood vessels and in individuals with moderate to heavy pigmentation. Use of optical particles doped with the FDA-approved near-infrared (NIR) absorber, indocyanine green (ICG), can potentially provide an effective method to overcome these limitations. Herein, we theoretically investigate the effectiveness of particles derived from erythrocytes, which contain ICG, in mediating photothermal destruction of PWS blood vessels. We refer to these particles as NIR erythrocyte-derived transducers (NETs). Our theoretical model consists of a Monte Carlo algorithm to estimate the volumetric energy deposition, a finite elements approach to solve the heat diffusion equation, and a damage integral based on an Arrhenius relationship to quantify tissue damage. The model geometries include simulated PWS blood vessels as well as actual human PWS blood vessels plexus obtained by the optical coherence tomography. Our simulation results indicate that blood vessels containing micron- or nano-sized NETs and irradiated at 755 nm have higher levels of photothermal damage as compared to blood vessels without NETs irradiated at 585 nm. Blood vessels containing micron-sized NETs also showed higher photothermal damage than blood vessels containing nano-sized NETs. The theoretical model presented can be used in guiding the fabrication of NETs with patient-specific optical properties to allow for personalized treatment based on the depth and size of blood vessels as well as the pigmentation of the individual’s skin.

Photoacoustic endoscopy offers in vivo examination of the visceral tissue using endogenous contrast, but its typical B-scan rate is ∼10  Hz, restricted by the speed of the scanning unit and the laser pulse repetition rate. Here, we present a transvaginal fast-scanning optical-resolution photoacoustic endoscope with a 250-Hz B-scan rate over a 3-mm scanning range. Using this modality, we not only illustrated the morphological differences of vasculatures among the human ectocervix, uterine body, and sublingual mucosa but also showed the longitudinal and cross-sectional differences of cervical vasculatures in pregnant women. This technology is promising for screening the visceral pathological changes associated with angiogenesis.

The Monte Carlo (MC) method is widely recognized as the gold standard for modeling light propagation inside turbid media. Due to the stochastic nature of this method, MC simulations suffer from inherent stochastic noise. Launching large numbers of photons can reduce noise but results in significantly greater computation times, even with graphics processing units (GPU)-based acceleration. We develop a GPU-accelerated adaptive nonlocal means (ANLM) filter to denoise MC simulation outputs. This filter can effectively suppress the spatially varying stochastic noise present in low-photon MC simulations and improve the image signal-to-noise ratio (SNR) by over 5 dB. This is equivalent to the SNR improvement of running nearly 3.5  ×   more photons. We validate this denoising approach using both homogeneous and heterogeneous domains at various photon counts. The ability to preserve rapid optical fluence changes is also demonstrated using domains with inclusions. We demonstrate that this GPU-ANLM filter can shorten simulation runtimes in most photon counts and domain settings even combined with our highly accelerated GPU MC simulations. We also compare this GPU-ANLM filter with the CPU version and report a threefold to fourfold speedup. The developed GPU-ANLM filter not only can enhance three-dimensional MC photon simulation results but also be a valuable tool for noise reduction in other volumetric images such as MRI and CT scans.

The scattering properties of biological tissue are highly dependent on the structure size, refractive index, and wavelength of the incident light. Furthermore, these scattering characteristics are strongly influenced by movements of the scattering objects. A method is developed to determine the angular- and spectral-resolved scattering properties that enabled the characterization of biological nano- and microscaled cell structures. Nanosecond pulses from a spectrally filtered supercontinuum light source are captured and time-resolved to depress background noise and minimize disruptive effects of the biological cells. The scattering characteristics of a monolayer of mouse fibroblast L929 cells are measured at defined wavelengths in a standard cell culture plate. Because of the size and distribution of the scattering structures, a Fourier transform-based Mie scattering scheme is used to analyze the data. The system is tested to detect structural changes of mouse fibroblast L929 cells before and after poisoning with Triton X100. The final result is the development of a contamination-free method to study pathological changes in cell cultures, necrosis, or other cell-damaging effects.

To characterize the optical clearing treatments in human colorectal tissues and possibly to differentiate between treatments of normal and pathological tissues, we have used a simple indirect method derived from Mie scattering theory to estimate the kinetics of the reduced scattering coefficient. A complementary method to estimate the kinetics of the scattering coefficient is also used so that the kinetics of the anisotropy factor and of the refractive index are also calculated. Both methods rely only on the thickness and collimated transmittance measurements made during treatment. The results indicate the expected time dependencies for the optical properties of both tissues: an increase in the refractive index and anisotropy factor and a decrease in the scattering coefficients. The similarity in the kinetics obtained for normal and pathological tissues indicates that optical clearing treatments can be applied also in pathological tissues to produce similar effects. The estimated time dependencies using experimental spectral data in the range from 400 to 1000 nm allowed us to compare the kinetics of the optical properties between different wavelengths.

Premature cervical remodeling is a critical precursor of spontaneous preterm birth, and the remodeling process is characterized by an increase in tissue hydration. Nevertheless, current clinical measurements of cervical remodeling are subjective and detect only late events, such as cervical effacement and dilation. Here, we present a photoacoustic endoscope that can quantify tissue hydration by measuring near-infrared cervical spectra. We quantify the water contents of tissue-mimicking hydrogel phantoms as an analog of cervical connective tissue. Applying this method to pregnant women in vivo, we observed an increase in the water content of the cervix throughout pregnancy. The application of this technique in maternal healthcare may advance our understanding of cervical remodeling and provide a sensitive method for predicting preterm birth.

While there exist many Monte Carlo (MC) programs for solving the radiative transfer equation (RTE) in biological tissues, we have identified a need for an open-source MC program that is sufficiently user-friendly for use in an education environment, in which detailed knowledge of compiling or UNIX command-line cannot be assumed. Therefore, we introduce MCmatlab, an open-source codebase thus far consisting of (a) a fast three-dimensional MC RTE solver and (b) a finite-element heat diffusion and Arrhenius-based thermal tissue damage simulator, both run in MATLAB. The kernel for both of these solvers is written in parallelized C and implemented as MATLAB MEX functions, combining the speed of C with the familiarity and versatility of MATLAB. We compare the RTE solver to Steven Jacques’ mcxyz, which it is inspired by, and present example results generated by the thermal model. MCmatlab is easy to install and use and can be used by students and experienced researchers alike for simulating tissue light propagation and, optionally, thermal damage.

Skin injury response to near-infrared (NIR) laser radiation between the minimum visible lesion threshold and ablation onset is not well understood. This study utilizes a 1070-nm diode-pumped Yb-fiber laser to explore the response of excised porcine skin to high-energy exposures in the suprathreshold injury region without inducing ablation. Concurrent high-speed videography is employed to determine a dichotomous response for three progressive damage categories: observable surface distortion, surface bubble formation due to contained intracutaneous water vaporization, and surface bubble rupture during exposure. Median effective dose (ED₅₀) values are calculated in these categories for 3- and 100-ms pulses with beam diameters (1  /  e²) of 3 mm (28, 35, and 49  J  /  cm²) and 7 mm (96, 141, and 212  J  /  cm²), respectively. Double-pulse cases are secondarily investigated. Experimental data are compared with the maximum permissible exposure limits and ablation onset simulated by a one-dimensional multiphysics model. Logistic regression analysis predicted injury events with ∼90  %   of accuracy. The distinction of skin response into progressive damage categories expands the current understanding of high-energy laser safety while underlining the unique biophysical effects during induced water phase change in tissue. These results prove to be useful in the diagnosis and treatment of NIR laser injuries.

Photoacoustic responses induced by laser-excited photothermal bubbles (PTBs) in colloidal gold solutions are relevant to the theranostics quality in biomedical applications. Confined to the complexity of nonstationary, multiscale events, and multiphysical parameters of PTBs, systematic studies of the photoacoustic effects remain obscure. Photoacoustic effects mediated by PTB dynamics and a physical mechanism are studied based on a proof-of-principle multimodal platform integrating side-scattering imaging, time-resolved optical response, and acoustic detection. Results show excitation energy, nanoparticle (NP) size, and NP concentration have strong influence on photoacoustic responses. Under the characteristic enhancement regime, the photoacoustic signal amplitude increases linearly with excitation energy and increases quadratically with the NP diameter. As for the effects of the NP concentration (characterized by absorption coefficient), a higher photoacoustic signal amplitude is generally induced by a dense NP distribution. However, with an increase in the NP size, the shielding effect of NP swarm prevents the increase of photoacoustic responses. This study presents experimental evidence of some key physical phenomena governing the PTB-induced photoacoustic signal generation in gold NP suspensions, which may help enrich theranostic approaches in clinical applications by rationalizing operation parameters.

Skin cancer is one of the most common cancers, including melanoma and nonmelanoma cancer. Melanoma can be easily detected by the observation of abnormal moles, but nonmelanoma signs and symptoms are not apparent in the early stages. We use the Stokes–Mueller matrix decomposition method to detect nonmelanoma at the early stage by decomposing the characteristics of polarized light interacting with normal and cancerous tissues. With this decomposition method, we extract nine optical parameters from biological tissues, namely the LB orientation angle (α), the LB phase retardance (β), the CB optical rotation angle (γ), the LD orientation angle (θ_d), the linear dichroism (D), the circular dichroism (R), the degrees of linear depolarization (e₁ and e₂), the degree of circular depolarization (e₃), and the depolarization index (Δ). The healthy skin and the induced nonmelanoma skin cancer of mice are analyzed and compared based on their optical parameters. We find distinctive ranges of values for normal skin tissue and nonmelanoma skin cancer, in which β and D in cancerous tissue are larger and nonmelanoma skin becomes less depolarized. This research creates an innovative solid foundation for the diagnosis of skin cancer in the future.

Determining the optical polarization properties of a skin lesion is a proposed method to differentiate melanoma from other skin lesions. We developed an in vivo Stokes polarimetry probe that fires a laser of known polarization at the skin and measures the Stokes parameters of the backscattered light in one shot. From these measured Stokes parameters, we can calculate the degree of polarization (DOP). Through testing on rough skin phantoms, a correlation between backscattered DOP and skin roughness was identified for both linear and circular input polarization, the latter of which was found to be more useful. In a pilot clinical trial of 69 skin lesions in vivo, it was found that the mean DOP for melanoma (linear input on melanoma: 0.46  ±  0.09) was greater than that of other lesions (linear input on all other lesions: 0.28  ±  0.01). This separation is greater for circular polarized input light, and it is likely that circular polarized light’s greater sensitivity to surface roughness contributes to this result. In addition, all skin lesions demonstrated a stronger depolarizing effect on circular polarized light than linear polarized light. We have identified DOP as a potentially useful measurement to identify melanoma among other types of skin lesions.

Sarcoidosis and tuberculoid leprosy (TL) are prototypes of granulomatous inflammation in dermatology, which embody one of the histopathology limitations in distinguishing some diseases. Recent advances in the use of nonlinear optical microscopy in skin have enabled techniques, such as second-harmonic generation (SHG), to become powerful tools to study the physical and biochemical properties of skin. We use SHG images to analyze the collagen network, to distinguish differences between sarcoidosis and TL granulomas. SHG images obtained from skin biopsies of 33 patients with TL and 24 with sarcoidosis retrospectively were analyzed using first-order statistics (FOS) and second-order statistics, such as gray-level co-occurrence matrix (GLCM). Among the four parameters evaluated (optical density, entropy, contrast, and second angular moment), only contrast demonstrated statistical significance, being higher in sarcoidosis (p  =  0.02; 4908.31 versus 2822.17). The results may indicate insufficient differentiating power for most tested FOS and GLCM parameters in classifying sarcoidosis and TL granulomas, when used individually. But in combination with histopathology (H&E and complementary stains, such as silver and fast acid stains), SHG analysis, like contrast, can contribute to distinguishing between these diseases. This study can provide a way to evaluate collagen distribution in granulomatous diseases.

Glycosaminoglycan (GAG) loss is an early marker of osteoarthritis, which is a clinical late stage disease that affects millions of people worldwide. The goal of our study was to evaluate the ability of a fiber-based fluorescence lifetime imaging (FLIm) technique to detect GAG loss in articular cartilage. Native bovine cartilage explants (n  =  20) were exposed to 0 (control), 0.5 (low), or 1  U  /  mL (high) concentrations of chondroitinase ABC (cABC) to create samples with different levels of GAG loss. FLIm assessment (excitation: 355 nm; detection: channel 1: 375 to 410 nm, channel 2: 450 to 485 nm, channel 3: 530 to 565 nm) was conducted on depth-resolved cross-sections of the cartilage sample. FLIm images, validated with histology, revealed that loss of GAG resulted in a decrease of fluorescence lifetime values in channel 2 (Δ  =  0.44  ns, p  <  0.05) and channel 3 (Δ  =  0.75  ns, p  <  0.01) compared to control samples (channel 2: 6.34 ns; channel 3: 5.22 ns). Fluorescence intensity ratio values were lower in channel 1 (37%, p  <  0.0001) and channel 2 (31% decrease, p  <  0.0001) and higher in channel 3 (23%, p  <  0.0001) relative to control samples. These results show that FLIm can detect the loss of GAG in articular cartilage and support further investigation into the feasibility of in vivo FLIm arthroscopy.

Impaired oxygen delivery and/or consumption in the retinal tissue underlies the pathophysiology of many retinal diseases. However, the essential tools for measuring oxygen concentration in retinal capillaries and studying oxygen transport to retinal tissue are still lacking. We show that two-photon phosphorescence lifetime microscopy can be used to map absolute partial pressures of oxygen (pO₂) in the retinal capillary plexus. Measurements were performed at various retinal depths in anesthetized mice under systemic normoxic and hyperoxic conditions. We used a newly developed two-photon phosphorescent oxygen probe, based on a two-photon absorbing platinum tetraphthalimidoporphyrin, and commercially available optics without correction for optical aberrations of the eye. The transverse and axial distances within the tissue volume were calibrated using a model of the eye’s optical system. We believe this is the first demonstration of in vivo depth-resolved imaging of pO₂ in retinal capillaries. Application of this method has the potential to advance our understanding of oxygen delivery on the microvascular scale and help elucidate mechanisms underlying various retinal diseases.

The imaging speed of optical resolution photoacoustic microscopy (OR-PAM) using pulsed excitation is fundamentally limited by the range ambiguity condition, which defines the maximum laser pulse repetition frequency (PRF). To operate at this theoretical upper limit and maximize acquisition speed, a custom-built fiber laser capable of operating at a PRF of up to 2 MHz was combined with a fast laser scanning optical OR-PAM system based on a stationary fiber-optic ultrasound sensor. A large area (10  mm  ×  10  mm) of the mouse ear was imaged within 8 s, when acquiring 16 million A-lines and operating the laser at a PRF of 2 MHz. This corresponds to a factor of four improvement in imaging speed compared to the fastest OR-PAM system previously reported. The ability to operate at high-imaging frame rates also allows the capture of hemodynamic events such as blood flow. It is considered that this system offers opportunities for high throughput imaging and visualizing dynamic physiological events using OR-PAM.

Frequency-doubled femtosecond Er-doped fiber laser is a low-cost and portable excitation source suitable for multiphoton endoscopy. The frequency-doubled wavelength at 780 nm is used to excite the intrinsic fluorescence signal. The frequency-doubling with a periodically poled MgO  :  LiNbO₃ (PPLN) is integrated in the distal end of the imaging head to achieve fiber connection. The imaging speed is further improved by optimizing the excitation laser source. A 0.3-mm length of PPLN crystal is selected and the Er-doped fiber laser is manipulated to match its bandwidth with the acceptance bandwidth of the PPLN. Through this optimization, a reduced pulsewidth of 80 fs of the frequency-doubled pulse is achieved. All-fiber dispersion compensation and pulse compression by single mode fiber is conducted, which makes the fiber laser directly fiber-coupled to the imaging head. An imaging speed of 4  frames  /  s is demonstrated on ex vivo imaging of unstained biological tissues, which is 10 times faster than our previous study using a 1-mm-long PPLN. The results show that miniature multiphoton endoscopy using frequency-doubled Er-doped fiber laser has great potential for clinical applications.

Free hemoglobin (FHB) concentration is considered a prospect quality indicator for erythrocyte suspensions (ES) under storage. Storage lesions alter the optical properties of ES and can be monitored by diffuse reflectance spectroscopy. Due to storage lesions, erythrocytes lyse and release hemoglobin into the extracellular medium. The purpose of the study is to model and assess the quality of ES units in a blood bank with diffuse reflectance measurements together with hematological variables reflecting absorption and scattering characteristics of ES. FHB concentrations were modeled based on the increased scattering in the extracellular medium. A semiempirical model was used for relating optical properties of ES to the diffuse reflectance measurements. The attenuation in the blood bag was computed and its influence was discarded via normalization, in accordance with Monte Carlo simulations. In the experiments, 40 ES units were measured multiple times during prolonged storage of 70 days. A generalized linear model was used for modeling the training set, and, in the validation, the highest correlation coefficient between predicted and actual FHB concentrations was 0.89. Predicting the actual value was accurate at a maximum level of R²  =  0.80. The error rate of the model in diagnosing the true quality was about 10%.

We report on the label-free real-time optical monitoring of DNA hybridization upon exposure to a flow of complementary DNA at different concentrations. The biosensor is composed of a silicon nitride integrated unbalanced Mach–Zehnder interferometer (MZI), with an integrated arrayed waveguide grating as a spectral filter. This MZI has been shown to have both sufficient multiplexing capability and limit of detection on the order of ^{10  −  6} RIU. Probe DNA, consisting of a 36-mer fragment is covalently immobilized on the silicon nitride integrated biosensor. The wavelength shift is monitored upon complementary DNA targets being flown over the sensor. Concentrations of 1 pM can be easily detected. Also, an alternative route to modify the sensor surface with carboxylic groups using the photochemical reaction of fatty acids is proposed and preliminary XPS results are presented. Moreover, preliminary results for DNA obtained from a rolling circle amplification (RCA-DNA) process and spiked in a realistic amplification buffer are presented.