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Significant numbers of liver biopsies fail to yield representative tissue samples. This study was conducted to evaluate the ability of LED-based diffuse reflectance spectroscopy to discriminate tumors from liver parenchyma. Ex vivo spectra were acquired from malignant lesions and liver parenchyma of 32 patients who underwent liver resection using a white light source and several LEDs. Integrated spectra of two combined LEDs with emission peaks at 470 nm and 515 nm were classified with 98.4% sensitivity and 99.2% specificity. The promising results could yield to a simple handheld and cost-efficient tool for real-time tissue differentiation implemented in a biopsy needle.
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In situ fluorescence lifetime imaging microscopy (FLIM) in an endoscopic configuration of the endogenous biomarker nicotinamide adenine dinucleotide (NADH) has a great potential for malignant tissue diagnosis. Moreover, two-photon nonlinear excitation provides intrinsic optical sectioning along with enhanced imaging depth. We demonstrate, for the first time to our knowledge, nonlinear endogenous FLIM in a fibered microscope with proximal detection, applied to NADH in cultured cells, as a first step to a nonlinear endomicroscope, using a double-clad microstructured fiber with convenient fiber length (> 3 m) and excitation pulse duration (≈50 fs). Fluorescence photons are collected by the fiber inner cladding and we show that its contribution to the impulse response function (IRF), which originates from its intermodal and chromatic dispersions, is small (< 600 ps) and stable for lengths up to 8 m and allows for short lifetime measurements. We use the phasor representation as a quick visualization tool adapted to the endoscopy speed requirements.
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We demonstrate a fiber source with the best performance from an ultrafast fiber oscillator to date. The ring-cavity Mamyshev oscillator produces ~50-nJ and ~40-fs pulses. The peak power is an order of magnitude higher than that of previous lasers with similar fiber mode area. This performance is achieved by designing the oscillator to support parabolic pulse formation which enables the management of unprecedented nonlinear phase shifts. Experimental results are limited by available pump power. Numerical simulations reveal key aspects of the pulse evolution, and realistically suggest that (after external compression) peak powers that approach 10 MW are possible from ordinary single-mode fiber. The combination of practical features such as environmental stability, established previously, with the performance described here make the Mamyshev oscillator extremely attractive for applications.
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In the present paper we show the optoacoustic (OA) response of two solutions of gold nanorods dispersed in distilled water (0.8 mg/ml) and hosted in tissue-like phantoms by using small arrays of high-power diode lasers [corrected] at 870 and 905 nm as excitation sources. The high-power diode lasers [corrected] are coupled to a 7-to-1 optical fiber bundle with output diameter of 675 µm. Each solution of gold nanorods exhibits an absorption peak close to the operating wavelength, i.e. ~860 nm and ~900 nm, respectively, to optimize the generation of OA signals. The phantoms are made of agar, intralipid and hemoglobin to simulate a soft biological tissue with reduced properties of scattering. Three 3-mm diameter tubes done in the phantoms at different depths (0.9 cm, 1.8 cm, and 2.7 cm) have been filled with gold nanorods. In this way, OA signals with appreciable SNR are generated at different depths in the phantoms. The high OA response exhibited by gold nanorods suggests their application in OA spectroscopy as exogenous contrast agents to detect and monitor emerging diseases like metastasis and arteriosclerotic plaques.
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[This corrects the article on p. 1430 in vol. 8, PMID: 28663839.].
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Successful breast conserving surgery consists of complete removal of the tumor while sparing healthy surrounding tissue. Despite currently available imaging and margin assessment tools, recognizing tumor tissue at a resection margin during surgery is challenging. Diffuse reflectance spectroscopy (DRS), which uses light for tissue characterization, can potentially guide surgeons to prevent tumor positive margins. However, inter-patient variation and changes in tissue physiology occurring during the resection might hamper this light-based technology. Here we investigate how inter-patient variation and tissue status (in vivo vs ex vivo) affect the performance of the DRS optical parameters. In vivo and ex vivo measurements of 45 breast cancer patients were obtained and quantified with an analytical model to acquire the optical parameters. The optical parameter representing the ratio between fat and water provided the best discrimination between normal and tumor tissue, with an area under the receiver operating characteristic curve of 0.94. There was no substantial influence of other patient factors such as menopausal status on optical measurements. Contrary to expectations, normalization of the optical parameters did not improve the discriminative power. Furthermore, measurements taken in vivo were not significantly different from the measurements taken ex vivo. These findings indicate that DRS is a robust technology for the detection of tumor tissue during breast conserving surgery.
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Photothermal therapy has shown to be a promising technique for local treatment of tumors. However, the main challenge for this technique is the availability of localized heat sources to minimize thermal damage in the surrounding healthy tissue. In this work, we demonstrate the use of optical fiber microheaters for inducing thermal lesions in soft tissue. The proposed devices incorporate carbon nanotubes or gold nanolayers on the tips of optical fibers for enhanced photothermal effects and heating of ex vivo biological tissues. We report preliminary results of small size photothermal lesions induced on mice liver tissues. The morphology of the resulting lesions shows that optical fiber microheaters may render useful for delivering highly localized heat for photothermal therapy.
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A dual-beam optical trap is used to trap and manipulate dielectric particles. When the refractive index of these particles is comparable to that of the surrounding medium, equilibrium trapping locations within the system shift from stable to unstable depending on fiber separation and particle size. This is due to to the relationship between gradient and scattering forces. We experimentally and computationally study the transitions between stable and unstable trapping of poly(methyl methacrylate) beads for a range of parameters relevant to experimental setups involving giant unilamellar vesicles. We present stability maps for various fiber separations and particle sizes, and find that careful attention to particle size and configuration is necessary to obtain reproducible quantitative results for soft matter stretching experiments.
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Optical tweezers play an important role in biological applications. However, it is difficult for traditional optical tweezers based on objective lenses to work in a three-dimensional (3D) solid far away from the substrate. In this work, we develop a fiber based optical trapping system, namely inclined dual fiber optical tweezers, that can simultaneously apply and measure forces both in water and in a 3D polyacrylamide gel matrix. In addition, we demonstrate in situ, non-invasive characterization of local mechanical properties of polyacrylamide gel by measurements on an embedded bead. The fiber optical tweezers measurements agree well with those of atomic force microscopy (AFM). The inclined dual fiber optical tweezers provide a promising and versatile tool for cell mechanics study in 3D environments.
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Early detection of structural or functional changes in dysplastic epithelia may be crucial for improving long-term patient care. Recent work has explored myriad non-invasive or minimally invasive "optical biopsy" techniques for diagnosing early dysplasia, such as high-resolution microendoscopy, a method to resolve sub-cellular features of apical epithelia, as well as broadband sub-diffuse reflectance spectroscopy, a method that evaluates bulk health of a small volume of tissue. We present a multimodal fiber-based microendoscopy technique that combines high-resolution microendoscopy, broadband (450-750 nm) sub-diffuse reflectance spectroscopy (sDRS) at two discrete source-detector separations (374 and 730 µm), and sub-diffuse reflectance intensity mapping (sDRIM) using a 635 nm laser. Spatial resolution, magnification, field-of-view, and sampling frequency were determined. Additionally, the ability of the sDRS modality to extract optical properties over a range of depths is reported. Following this, proof-of-concept experiments were performed on tissue-simulating phantoms made with poly(dimethysiloxane) as a substrate material with cultured MDA-MB-468 cells. Then, all modalities were demonstrated on a human melanocytic nevus from a healthy volunteer and on resected colonic tissue from a murine model. Qualitative in vivo image data is correlated with reduced scattering and absorption coefficients.
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A single human red blood cell was optically stretched along two counter-propagating fiber-optic Bessel-like beams in an integrated lab-on-a-chip structure. The beam enabled highly localized stretching of RBC, and it induced a nonlinear mechanical deformation to finally reach an irreversible columnar shape that has not been reported. We characterized and systematically quantified this optically induced mechanical deformation by the geometrical aspect ratio of stretched RBC and the irreversible stretching time. The proposed RBC mechanism can realize a versatile and compact opto-mechanical platform for optical diagnosis of biological substances in the single cell level.
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We demonstrate a miniaturized single beam fiber optical trapping probe based on a high numerical aperture graded index (GRIN) micro-objective lens. This enables optical trapping at a distance of 200µm from the probe tip. The fiber trapping probe is characterized experimentally using power spectral density analysis and an original approach based on principal component analysis for accurate particle tracking. Its use for biomedical microscopy is demonstrated through optically mediated immunological synapse formation.
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In this paper, a textile-based sensing principle for long term photopletysmography (PPG) monitoring is presented. Optical fibers were embroidered into textiles such that out-coupling and in-coupling of light was possible. The "light-in light-out" properties of the textile enabled the spectroscopic characterization of human tissue. For the optimization of the textile sensor, three different carrier fabrics and different fiber modifications were compared. The sample with best light coupling efficiency was successfully used to measure heart rate and SpO2 values of a subject. The latter was determined by using a modified Beer-Lambert law and measuring the light attenuation at two different wavelengths (632 nm and 894 nm). Moreover, the system was adapted to work in reflection mode which makes the sensor more versatile. The measurements were additionally compared with commercially available system and showed good correlation.
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The detailed mechanisms associated with the influence of scattering and absorption properties on the fluorescence intensity sampled by a single optical fiber have recently been elucidated based on Monte Carlo simulated data. Here we develop an experimental single fiber fluorescence (SFF) spectroscopy setup and validate the Monte Carlo data and semi-empirical model equation that describes the SFF signal as a function of scattering. We present a calibration procedure that corrects the SFF signal for all system-related, wavelength dependent transmission efficiencies to yield an absolute value of intrinsic fluorescence. The validity of the Monte Carlo data and semi-empirical model is demonstrated using a set of fluorescent phantoms with varying concentrations of Intralipid to vary the scattering properties, yielding a wide range of reduced scattering coefficients (µ's = 0-7 mm (-1)). We also introduce a small modification to the model to account for the case of µ's = 0 mm (-1) and show its relation to the experimental, simulated and theoretically calculated value of SFF intensity in the absence of scattering. Finally, we show that our method is also accurate in the presence of absorbers by performing measurements on phantoms containing red blood cells and correcting for their absorption properties.
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We present a novel algorithm to detect contact with tissue and automate data acquisition. Contact fiber-optic probe systems are useful in noninvasive applications and real-time analysis of tissue properties. However, applications of these technologies are limited to procedures with visualization to ensure probe-tissue contact and individual user techniques can introduce variability. The software design exploits the system previously designed by our group as an optical method to automatically detect tissue contact and trigger acquisition. This method detected tissue contact with 91% accuracy, detected removal from tissue with 83% accuracy and reduced user variability by > 8%. Without the need for additional hardware, this software algorithm can easily integrate into any fiber-optic system and expands applications where visualization is difficult.
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Multi diameter single fiber reflectance (MDSFR) spectroscopy is a non-invasive optical technique based on using multiple fibers of different diameters to determine both the reduced scattering coefficient (µs') and a parameter γ that is related to the angular distribution of scattering, where γ = (1-g2)/(1-g1) and g1 and g2 the first and second moment of the phase function, respectively. Here we present the first in vivo MDSFR measurements of µs'(λ) and γ(λ) and their wavelength dependence. MDSFR is performed on nineteen mice in four tissue types including skin, liver, normal tongue and in an orthotopic oral squamous cell carcinoma. The wavelength-dependent slope of µs'(λ) (scattering power) is significantly higher for tongue and skin than for oral cancer and liver. The reduced scattering coefficient at 800 nm of oral cancer is significantly higher than of normal tongue and liver. Gamma generally increases with increasing wavelength; for tumor it increases monotonically with wavelength, while for skin, liver and tongue γ(λ) reaches a plateau or even decreases for longer wavelengths. The mean γ(λ) in the wavelength range 400-850 nm is highest for liver (1.87 ± 0.07) and lowest for skin (1.37 ± 0.14). Gamma of tumor and normal tongue falls in between these values where tumor exhibits a higher average γ(λ) (1.72 ± 0.09) than normal tongue (1.58 ± 0.07). This study shows the potential of using light scattering spectroscopy to optically characterize tissue in vivo.
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Quantitative determination of fluorophore content from fluorescence measurements in turbid media, such as tissue, is complicated by the influence of scattering properties on the collected signal. This study utilizes a Monte Carlo model to characterize the relationship between the fluorescence intensity collected by a single fiber optic probe (F(SF)) and the scattering properties. Simulations investigate a wide range of biologically relevant scattering properties specified independently at excitation (λ(x)) and emission (λ(m)) wavelengths, including reduced scattering coefficients in the range µ'(s)(λ(x)) ∈ [0.1 - 8]mm(-1) and µ'(s)(λ(m)) ∈ [0.25 - 1] × µ'(s)(λ(x)). Investigated scattering phase functions (P(θ)) include both Henyey-Greenstein and Modified Henyey-Greenstein forms, and a wide range of fiber diameters (d(f) ∈ [0.2 - 1.0] mm) was simulated. A semi-empirical model is developed to estimate the collected F(SF) as the product of an effective sampling volume, and the effective excitation fluence and the effective escape probability within the effective sampling volume. The model accurately estimates F(SF) intensities (r=0.999) over the investigated range of µ'(s)(λ(x)) and µ'(s)(λ(m)), is insensitive to the form of the P(θ), and provides novel insight into a dimensionless relationship linking F(SF) measured by different d(f).
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This paper presents a relationship between the intensity collected by a single fiber reflectance device (R(SF)) and the fiber diameter (d(fib)) and the reduced scattering coefficient ( µs') and phase function (p(θ)) of a turbid medium. Monte Carlo simulations are used to identify and model a relationship between R(SF) and dimensionless scattering ( µs'dfib). For µs'dfib > 10 we find that R(SF) is insensitive to p(θ). A solid optical phantom is constructed with µs' ≈ 220 mm-1 and is used to convert R(SF) of any turbid medium to an absolute scale. This calibrated technique provides accurate estimates of µs' over a wide range ([0.05 - 8] mm(-1)) for a range of d(fib) ([0.2 - 1] mm).