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We report the design, development, and characterization of a novel multi-spectral fluorescence lifetime measurement device incorporating solid-state detectors and automated gain control. For every excitation pulse (∼1 µJ, 600 ps), this device records complete fluorescence decay from multiple spectral channels simultaneously within microseconds, using a dedicated UV enhanced avalanche photodetector and analog to digital convert (2.5 GS/s) in each channel. Fast (

We report the design, development, and characterization of the first multispectral fluorescence lifetime imaging platform designed for clinical application based on avalanche photodiodes (APD). Consistent and accurate measurement of fluorescence lifetimes is achieved. Fast dynamic gain adjustment via APD bias modulation is implemented independently for each spectral channel, maximizing photon economy and greatly improving signal quality. Compared to conventional MCP-PMT based systems, the new platform presents 5-fold reduction of lifetime measurement variability under identical experiment conditions. This improved photon economy leads to 35 times faster coverage of imaging field in clinical practice with identical tissue exposure, making the new platform better suited for clinical application.

Endogenous and exogenous fluorescence emission from biological samples encodes complementary information. Here we report, to the best of our knowledge, the first results from an optical imaging platform with interleaved excitation and detection of exogenous and endogenous fluorescence from tissue samples using a single flexible multimode fiber that delivers the excitation beam and collects the emitted light. A custom-built reflective optical chopper wheel with synchronized rotation temporally multiplexes an autofluorescence lifetime imaging apparatus with an intensity-based fluorescence module tailored to imaging green fluorescent protein. We demonstrate the functionality of such platform imaging dyes of varying fluorescence signatures and resolving cellularized areas on bio-engineered tissue constructs.

Due to loss of tactile feedback the assessment of tumor margins during robotic surgery is based only on visual inspection, which is neither significantly sensitive nor specific. Here we demonstrate time-resolved fluorescence spectroscopy (TRFS) as a novel technique to complement the visual inspection of oral cancers during transoral robotic surgery (TORS) in real-time and without the need for exogenous contrast agents. TRFS enables identification of cancerous tissue by its distinct autofluorescence signature that is associated with the alteration of tissue structure and biochemical profile. A prototype TRFS instrument was integrated synergistically with the da Vinci Surgical robot and the combined system was validated in swine and human patients. Label-free and real-time assessment and visualization of tissue biochemical features during robotic surgery procedure, as demonstrated here, not only has the potential to improve the intraoperative decision making during TORS but also other robotic procedures without modification of conventional clinical protocols.

We present a scheme for conversion of pulsed light from the infrared to the red spectral region, using an aperiodically poled ferroelectric crystal within a resonant cavity in which two cascaded nonlinear optical processes occur when pumped with a pulsed Nd:YAG laser. This device emits 9 ns pulses of over 1 mJ at 710 nm and is a viable source for future biomedical applications. All Rights Reserved © 2015 Universidad Nacional Autonoma de Mexico, Centro de Ciencias Aplicadas y Desarrollo Tecnologico. This is an open access item distributed under the Creative Commons CC License BY-NC-ND 4.0.

Fiber-based imaging of tissue autofluorescence using ultraviolet (UV) excitation is a highly flexible tool used to probe structure and composition. In this Letter, we report, to the best of our knowledge, the first results from a single-fiber imaging system employing a custom double-clad fiber to acquire multispectral fluorescence lifetime images at two distinct spatial resolutions. We characterize the lateral point spread function and fluorescent background of the system and show how enhanced resolution can identify features such as trabeculae in ex vivo murine bone samples.

Fluorescence lifetime technique has demonstrated potential for analysis of atherosclerotic lesions and for complementing existing intravascular imaging modalities such as intravascular ultrasound (IVUS) in identifying lesions at high risk of rupture. This study presents a multimodal catheter system integrating a 40 MHz commercial IVUS and fluorescence lifetime imaging (FLIm) using fast helical motion scanning (400 rpm, 0.75 mm/s), able to acquire in vivo in pulsatile blood flow the autofluorescence emission of arterial vessels with high precision (5.08 ± 0.26 ns mean average lifetime over 13 scans). Co-registered FLIm and IVUS data allowed 3D visualization of both biochemical and morphological vessel properties. Current study supports the development of clinically compatible intravascular diagnostic system integrating FLIm and demonstrates, to our knowledge, the first in vivo intravascular application of a fluorescence lifetime imaging technique. Keywords: time resolved fluorescence spectroscopy, fluorescence lifetime imaging, intravascular ultrasound (IVUS), intravascular catheter, atherosclerosis 1. Introduction Coronary artery disease is the leading cause of death in western societies [1]. Although many diagnostic tools have been developed to help predict which patients will suffer cardiovascular complications, current diagnostic techniques have significant limitations [2]. New techniques, including intravascular methods, are needed in order to make a significant impact on this public health challenge. Point time-resolved fluorescence spectroscopy (TRFS) and fluorescence lifetime imaging microscopy (FLIM) have demonstrated ability to assess a plurality of diagnostic features associated with arterial wall pathologies including atherosclerotic plaques at risk of rupture [3]. However, the overall TRFS and FLIM potential for atherosclerotic plaques analysis was primarily evaluated ex-vivo in arterial tissue specimens with intima directly exposed to the fiberoptic probe interrogation. Practical intravascular application of these techniques requires development of TRFS systems and catheters capable of rapid acquisition of robust time-resolved fluorescence data in vivo from the luminal surface of the artery in the presence of pulsa-tile blood flow. A novel implementation of a TRFS apparatus with the ability to rapidly (

The molecular basis of nonlinear optical (NLO) chiral effects in the amide I region of type I collagen was investigated using sum-frequency generation vibrational spectroscopy; chiral and achiral tensor elements were separated using different input/output beam polarization conditions. Spectra were obtained from native rat tail tendon (RTT) collagen and from cholesteric liquid crystal-like (LC) type I collagen films. Although RTT and LC collagen both possess long-range order, LC collagen lacks the complex hierarchical organization of RTT collagen. Their spectra were compared to assess the role of such organization in NLO chirality. No significant differences were observed between RTT and LC with respect to chiral or achiral spectra. These findings suggest that amide I NLO chiral effects in type I collagen assemblies arise predominantly from the chiral organization of amide chromophores within individual collagen molecules, rather than from supramolecular structures. The study suggests that sum-frequency generation vibrational spectroscopy may be uniquely valuable in exploring fundamental aspects of chiral nonlinearity in complex macromolecular structures.

The progression of atherosclerosis in coronary vessels involves distinct pathological changes in the vessel wall. These changes manifest in the formation of a variety of plaque sub-types. The ability to detect and distinguish these plaques, especially thin-cap fibroatheromas (TCFA) may be relevant for guiding percutaneous coronary intervention as well as investigating new therapeutics. In this work we demonstrate the ability of fluorescence lifetime imaging (FLIm) derived parameters (lifetime values from sub-bands 390/40 nm, 452/45 nm and 542/50 nm respectively) for generating classification maps for identifying eight different atherosclerotic plaque sub-types in ex vivo human coronary vessels. The classification was performed using a support vector machine based classifier that was built from data gathered from sixteen coronary vessels in a previous study. This classifier was validated in the current study using an independent set of FLIm data acquired from four additional coronary vessels with a new rotational FLIm system. Classification maps were compared to co-registered histological data. Results show that the classification maps allow identification of the eight different plaque sub-types despite the fact that new data was gathered with a different FLIm system. Regions with diffuse intimal thickening (n=10), fibrotic tissue (n=2) and thick-cap fibroatheroma (n=1) were correctly identified on the classification map. The ability to identify different plaque types using FLIm data alone may serve as a powerful clinical and research tool for studying atherosclerosis in animal models as well as in humans.

Fluorescence lifetime imaging has been shown to be a robust technique for biochemical and functional characterization of tissues and to present great potential for intraoperative tissue diagnosis and guidance of surgical procedures. We report a technique for real-time mapping of fluorescence parameters (i.e. lifetime values) onto the location from where the fluorescence measurements were taken. This is achieved by merging a 450 nm aiming beam generated by a diode laser with the excitation light in a single delivery/collection fiber and by continuously imaging the region of interest with a color CMOS camera. The interrogated locations are then extracted from the acquired frames via color-based segmentation of the aiming beam. Assuming a Gaussian profile of the imaged aiming beam, the segmentation results are fitted to ellipses that are dynamically scaled at the full width of three automatically estimated thresholds (50%, 75%, 90%) of the Gaussian distribution's maximum value. This enables the dynamic augmentation of the white-light video frames with the corresponding fluorescence decay parameters. A fluorescence phantom and fresh tissue samples were used to evaluate this method with motorized and hand-held scanning measurements. At 640x512 pixels resolution the area of interest augmented with fluorescence decay parameters can be imaged at an average 34 frames per second. The developed method has the potential to become a valuable tool for real-time display of optical spectroscopy data during continuous scanning applications that subsequently can be used for tissue characterization and diagnosis.

Fiber based fluorescence lifetime imaging has shown great potential for intraoperative diagnosis and guidance of surgical procedures. Here we describe a novel method addressing a significant challenge for the practical implementation of this technique, i.e., the real-time display of the quantified biochemical or functional tissue properties superimposed on the interrogated area. Specifically, an aiming beam (450 nm) generated by a continuous-wave laser beam was merged with the pulsed fluorescence excitation light in a single delivery/collection fiber and then imaged and segmented using a color-based algorithm. We demonstrate that this approach enables continuous delineation of the interrogated location and dynamic augmentation of the acquired frames with the corresponding fluorescence decay parameters. The method was evaluated on a fluorescence phantom and fresh tissue samples. Current results demonstrate that 34 frames per second can be achieved for augmenting videos of 640 $\times $ 512 pixels resolution. Also we show that the spatial resolution of the fluorescence lifetime map depends on the tissue optical properties, the scanning speed, and the frame rate. The dice similarity coefficient between the fluorescence phantom and the reconstructed maps was estimated to be as high as 93%. The reported method could become a valuable tool for augmenting the surgeon's field of view with diagnostic information derived from the analysis of fluorescence lifetime data in real-time using handheld, automated, or endoscopic scanning systems. Current method provides also a means for maintaining the tissue light exposure within safety limits. This study provides a framework for using an aiming beam with other point spectroscopy applications.

Detection of atherosclerotic plaque vulnerability has critical clinical implications for avoiding sudden death in patients with high risk of plaque rupture. We report on multimodality imaging of ex-vivo human carotid plaque samples using a system that integrates fluorescence lifetime imaging (FLIM), ultrasonic backscatter microscopy (UBM), and photoacoustic imaging (PAI). Biochemical composition is differentiated with a high temporal resolution and sensitivity at the surface of the plaque by the FLIM subsystem. 3D microanatomy of the whole plaque is reconstructed by the UBM. Functional imaging associated with optical absorption contrast is evaluated from the PAI component. Simultaneous recordings of the optical, ultrasonic, and photoacoustic data present a wealth of complementary information concerning the plaque composition, structure, and function that are related to plaque vulnerability. This approach is expected to improve our ability to study atherosclerotic plaques. The multimodal system presented here can be translated into a catheter based intraluminal system for future clinical studies.

To explore integrated solar energy harvesting as a power source for low power systems, an array of energy scavenging photodiodes based on a passive-pixel architecture for CMOS imagers has been fabricated together with storage capacitors implemented using on-chip interconnect in a 0.35-mum bulk process. Integrated vertical plate capacitors enable dense energy storage without limiting optical efficiency. Tests were conducted with both a white light source and a green laser. Measurements indicate that 225 muW/mm2 output power may be generated by white light with an intensity of 20 kLUX.

We report a novel technique for continuous acquisition, processing and display of fluorescence lifetimes enabling real-time tissue diagnosis through a single hand held or biopsy fiber-optic probe. A scanning multispectral time-resolved fluorescence spectroscopy (ms-TRFS) with self-adjustable photon detection range was developed to account for the dynamic changes of fluorescence intensity typically encountered in clinical application. A fast algorithm was implemented in the ms-TRFS software platform, providing up to 15 Hz continuous display of fluorescence lifetime values. Potential applications of this technique, including biopsy guidance, and surgical margins delineation were demonstrated in proof-of-concept experiments. Current results showed accurate display of fluorescence lifetimes values and discrimination of distinct fluorescence markers and tissue types in real-time (< 100 ms per data point).

We report a scanning imaging system that enables high speed multispectral fluorescence lifetime imaging (FLIm) of coronary arteries. This system combines a custom low profile (3 Fr) imaging catheter using a 200 μm core side viewing UV-grade silica fiber optic, an acquisition system able to measure fluorescence decays over four spectral bands at 20 kHz and a fast data analysis and display module. In vivo use of the system has been optimized, with particular emphasis on clearing blood from the optical pathway. A short acquisition time (5 seconds for a 20 mm long coronary segment) enabled data acquisition during a bolus saline solution injection through the 7 Fr catheter guide. The injection parameters were precisely controlled using a power injector and optimized to provide good image quality while limiting the bolus injection duration and volume (12 cc/s, 80 cc total volume). The ability of the system to acquire data in vivo was validated in healthy swine by imaging different sections of the left anterior descending (LAD) coronary. A stent coated with fluorescent markers was placed in the LAD and imaged, demonstrating the ability of the system to discriminate in vivo different fluorescent features and structures from the vessel background fluorescence using spectral and lifetime information. Intensity en face images over the four bands of the instrument were available within seconds whereas lifetime images were computed in 2 minutes, providing efficient feedback during the procedure. This successful demonstration of FLIm in coronaries enables future study of atherosclerotic cardiovascular diseases.

Fluorescence lifetime imaging (FLIM) has demonstrated potential for robust assessment of atherosclerotic plaques biochemical composition and for complementing conventional intravascular ultrasound (IVUS), which provides information on plaque morphology. The success of such a bi-modal imaging modality depends on accurate segmentation of the IVUS images and proper angular registration between these two modalities. This paper reports a novel IVUS segmentation methodology addressing this issue. The image preprocessing consisted of denoising, using the Wiener filter, followed by image smoothing, implemented through the application of the alternating sequential filter on the edge separability metric images. Extraction of the lumen/intima and media/adventitia boundaries was achieved by tracing the gray-scale peaks over the A-lines of the IVUS preprocessed images. Cubic spline interpolation, in both cross-sectional and longitudinal directions, ensured boundary smoothness and continuity. The detection of the guide-wire artifact in both modalities is used for angular registration. Intraluminal studies were conducted in 13 ex vivo segments of human coronaries. The IVUS segmentation accuracy was assessed against independent manual tracings, providing 91.82% sensitivity and 97.55% specificity. The proposed methodology makes the bi-modal FLIM and IVUS approach feasible for comprehensive intravascular diagnosis by providing co-registered biochemical and morphological information of atherosclerotic plaques.

Vinyl-addition poly(norbornene) copolymers functionalized with nonlinear optical chromophore side groups have been prepared using (η6-toluene)Ni(C6F5)2, and their electrooptic properties have been characterized. The nickel complex used to polymerize the norbornene monomers is tolerant to many functional groups found in nonlinear optical chromophores although nitriles and amines other than trisubstituted amines strongly inhibit the reaction. A vinyl-addition copolymer of hexylnorbornene and a norbornene-functionalized Disperse Red 1 chromophore was scaled up and studied in detail. Initial studies indicate that electric field poling is effective but that relaxation of polar order in the poly(norbornene) is faster than in a comparable methacrylate copolymer.

We report the development and validation of a hybrid intravascular diagnostic system combining multispectral fluorescence lifetime imaging (FLIm) and intravascular ultrasound (IVUS) for cardiovascular imaging applications. A prototype FLIm system based on fluorescence pulse sampling technique providing information on artery biochemical composition was integrated with a commercial IVUS system providing information on artery morphology. A customized 3-Fr bimodal catheter combining a rotational side-view fiberoptic and a 40-MHz IVUS transducer was constructed for sequential helical scanning (rotation and pullback) of tubular structures. Validation of this bimodal approach was conducted in pig heart coronary arteries. Spatial resolution, fluorescence detection efficiency, pulse broadening effect, and lifetime measurement variability of the FLIm system were systematically evaluated. Current results show that this system is capable of temporarily resolving the fluorescence emission simultaneously in multiple spectral channels in a single pullback sequence. Accurate measurements of fluorescence decay characteristics from arterial segments can be obtained rapidly (e.g., 20 mm in 5 s), and accurate co-registration of fluorescence and ultrasound features can be achieved. The current finding demonstrates the compatibility of FLIm instrumentation with in vivo clinical investigations and its potential to complement conventional IVUS during catheterization procedures.

The risk of atherosclerosis plaque rupture cannot be assessed by the current imaging systems and thus new multi-modal technologies are under investigation. This includes combining a new fluorescence lifetime imaging (FLIm) technique, which is sensitive to plaque biochemical features, with conventional intravascular ultrasound (IVUS), which provides information on plaque morphology. In this study we present an automated method allowing for the co-registration of imaging data acquired based on these two techniques. Intraluminal studies were conducted in ex-vivo segments of human coronaries with a multimodal catheter integrating a commercial IVUS (40 MHz) and a rotational side-viewing fiber based multispectral FLIm system (355 nm excitation, 390±20, 452±22 and 542±25 nm acquisition wavelengths). The proposed method relies on the lumen/intima boundary extraction from the IVUS polar images. Image restoration is applied for the noise reduction and edge enhancement, while gray-scale peak tracing over the A-lines of the IVUS polar images is applied for the lumen boundary extraction. The detection of the guide-wire artifact is used for the angular registration between FLIm and IVUS data, after which the lifetime values can be mapped onto the segmented lumen/intima interface. The segmentation accuracy has been assessed against manual tracings, providing 0.120±0.054 mm mean Hausdorff distance. This method makes the bi-modal FLIm and IVUS approach feasible for comprehensive intravascular diagnostic by providing co-registered biochemical and morphological information about atherosclerotic plaques.

We present the development of a source of deep-red radiation for photoacoustic imaging. This source, which is based on two cascaded wavelength conversion processes in aperiodically poled lithium niobate, emits 10 nanosecond pulses of over 500 µJ at 710 nm. Photoacoustic images were obtained from phantoms designed to mimic the optical and acoustic properties of oral tissue. Results indicate this device is a viable source of optical pulses for photoacoustic applications.

Electrooptic modulators that consist of an optical fiber waveguide coupled to an electrooptic waveguide are reviewed. Desirable attributes of these devices are that the optical fiber is uninterrupted and the interaction with the electrooptic region occurs only where the optical properties are modulated. In this paper we review in-line fiber evanescent field modulators that we have implemented with electrooptic polymers and compound semiconductor quantum wells. We show that the beam propagation method can accurately simulate the behavior measured in these devices.

The transfer function of an external modulator is the critical factor that determines the spurious free dynamic range, signal power gain, and noise figure of a wide-band analog RF-photonic link. We present an in-line asymmetric directional coupler modulator with distributed optical loss capable of providing a transfer function linearized to the fourth order for multioctave bandwidths. This modulator compares favorably with multiple modulator suboctave linearization techniques consisting of series symmetric directional coupler and series Mach-Zehnder modulators and superoctave parallel Mach-Zehnder, and cascaded sections containing series Mach-Zehndersymmetric directional coupler modulators. The optimum bias point for the asymmetric directional coupler modulator is determined by device dimensions and material parameters with relative insensitivity to errors introduced during fabrication. Optical insertion loss imposes significant limitations on modulator performance. An implementation of an in-line asymmetric directional coupler modulator is discussed that eliminates excess loss due to fiber coupling in order to achieve a large RF power gain and competitive noise figure compared to the linearization configurations.

This work reports a multimodal system for label-free tissue diagnosis combining fluorescence lifetime imaging (FLIm), ultrasound backscatter microscopy (UBM), and photoacoustic imaging (PAI). This system provides complementary biochemical, structural and functional features allowing for enhanced in vivo detection of oral carcinoma. Results from a hamster oral carcinoma model (normal, precancer and carcinoma) are presented demonstrating the ability of FLIm to delineate biochemical composition at the tissue surface, UBM and related radiofrequency parameters to identify disruptions in the tissue microarchitecture and PAI to map optical absorption associated with specific tissue morphology and physiology.

Fluorescence lifetime technique has demonstrated potential for analysis of atherosclerotic lesions and for complementing existing intravascular imaging modalities such as intravascular ultrasound (IVUS) in identifying lesions at high risk of rupture. This study presents a multimodal catheter system integrating a 40 MHz commercial IVUS and fluorescence lifetime imaging (FLIm) using fast helical motion scanning (400 rpm, 0.75 mm/s), able to acquire in vivo in pulsatile blood flow the autofluorescence emission of arterial vessels with high precision (5.08 ± 0.26 ns mean average lifetime over 13 scans). Co-registered FLIm and IVUS data allowed 3D visualization of both biochemical and morphological vessel properties. Current study supports the development of clinically compatible intravascular diagnostic system integrating FLIm and demonstrates, to our knowledge, the first in vivo intravascular application of a fluorescence lifetime imaging technique.

Tissue diagnostic features generated by a bimodal technique integrating scanning time-resolved fluorescence spectroscopy (TRFS) and ultrasonic backscatter microscopy (UBM) are investigated in an in vivo hamster oral carcinoma model. Tissue fluorescence is excited by a pulsed nitrogen laser and spectrally and temporally resolved using a set of filters/dichroic mirrors and a fast digitizer, respectively. A 41-MHz focused transducer (37-μm axial, 65-μm lateral resolution) is used for UBM scanning. Representative lesions of the different stages of carcinogenesis show that fluorescence characteristics complement ultrasonic features, and both correlate with histological findings. These results demonstrate that TRFS-UBM provide a wealth of co-registered, complementary data concerning tissue composition and structure as it relates to disease status. The direct co-registration of the TRFS data (sensitive to surface molecular changes) with the UBM data (sensitive to cross-sectional structural changes and depth of tumor invasion) is expected to play an important role in pre-operative diagnosis and intra-operative determination of tumor margins.

Photoswitchable spiropyran has been conjugated to the crowned ring system DO3A, which improves its solubility in dipolar and polar media and stabilizes the merocyanine isomer. Adding the lanthanide ion gadolinium(III) to the macrocyclic ring system leads to a photoresponsive magnetic resonance imaging contrast agent that displays an increased spin-lattice relaxation time (T1) upon visible light stimulation. In this work, the photoresponse of this photochromic molecule to weak light illumination using blue and green light emitting diodes was investigated, simulating the emission spectra from bioluminescent enzymes. Photon emission rate of the light emitting diodes was changed, from 1.75 × 1016 photons·s−1 to 2.37 × 1012 photons·s−1. We observed a consistent visible light-induced isomerization of the merocyanine to the spiropyran form with photon fluxes as low as 2.37 × 1012 photons·s−1 resulting in a relaxivity change of the compound. This demonstrates the potential for use of the described imaging probes in low light level applications such as sensing bioluminescence enzyme activity. The isomerization behavior of gadolinium(III)-ion complexed and non-complexed spiropyran-DO3A was analyzed in water and ethanol solution in response to low light illumination and compared to the emitted photon emission rate from over-expressed Gaussia princeps luciferase.

This study describes a scanning time-resolved fluorescence spectroscopy (TRFS) system designed to continuously acquire fluorescence emission and to reconstruct fluorescence lifetime images (FLIM) from a luminal surface by using a catheter-based optical probe with rotary joint and pull-back device. The ability of the system to temporally and spectrally resolve the fluorescence emission from tissue was validated using standard dyes and tissue phantoms (e.g., ex vivo pig aorta phantom). Current results demonstrate that this system is capable to reliably resolve the fluorescence emission of multiple fluorophores located in the lumen; and suggest its potential for intravascular detection of distinct biochemical features of atherosclerotic plaques.

We describe the fabrication and performance of a thin-film electro-optic Fabry-Perot modulator operating in reflection, at wavelengths compatible with silicon detectors, fabricated on a silicon substrate. A nonlinear polymer is used with a low dielectric constant that permits an electrical bandwidth in excess of 100 MHz with low modulating energy. Existing thin-film fabrication techniques and materials are used to demonstrate device performance. We report modulation of approximately 1.5% and describe foreseen improvements in nonlinear polymers and technology which promise in excess of 5% modulation with a 5 V modulating voltage. >

The authors describe a self-homodyne technique to measure the modulation efficiency of an optical phase modulator. The phase modulator is driven by a sine wave and the baseband spectrum is measured and analyzed. Combinations of the ratio of harmonics are used to estimate V/sub /spl pi// of a commercially available modulator with a resolution of 0.8%. The method offers simplicity because it uses a single laser, does not require sensitive optical adjustments, and lends itself to the measurement of V/sub /spl pi// as a function of modulation frequency and optical carrier wavelength.

This paper presents an endoscopic configuration for measurements of tissue autofluorescence using two-photon excitation and time-correlated single photon counting detection through a double-clad photonic crystal fiber (DC-PCF) without pre-chirping of laser pulses. The instrument performance was evaluated by measurements of fluorescent standard dyes, biological fluorophores (collagen and elastin), and tissue specimens (muscle, cartilage, tendon). Current results demonstrate the ability of this system to accurately retrieve the fluorescence decay profile and lifetime of these samples. This simple setup, which offers larger penetration depth than one-photon-based techniques, may be combined with morphology-yielding techniques such as photoacoustic and ultrasound imaging.

We report the development and validation of an intravascular rotary catheter that enables bi-modal interrogation of arterial pathologies based on fast-frame time-resolved fluorescence spectroscopy (TRFS) and intravascular ultrasound (IVUS). The catheter is based on a parallel design that allows for independent rotation of the ultrasonic and optical channels within an 8 Fr outer diameter catheter sheath and integrates a low volume flushing channel for blood removal in the optical pathways. In current configuration, the two channels consist of a) a standard 8 Fr IVUS catheter with single element transducer (15 MHz) and b) a side-viewing UV-grade silica/silica fiber optic (400 μm core). The catheter is terminated by a small (0.82 mm internal diameter) polyimide tube to keep the fiber stable within the sheath. To clear the field of view from blood, a saline solution can be flushed in a sheath channel, concentric with the fiber optic, through the tube and in a radial opening aligned with the fiber's optical beam. The flushing function was optimized with a computational fluid dynamics (CFD) model pursued in a parallel study. The ability of the catheter to operate in intraluminal setting in blood flow, the effect of probe-to-tissue distance on optical signal and ability to generate co-registered TRFS and IVUS data were demonstrated in blood vessel phantoms. Current results demonstrate the feasibility of the described catheter for parallel interrogation of vessel walls based on TRFS and IVUS and to generate robust TRFS data. These results facilitate further development of a bi-modal TRFS-IVUS technique for intravascular diagnosis of atherosclerotic cardiovascular diseases including vulnerable plaques.

A 2.5- mu m-thick corona-poled copolymer film was used to double approximately 13-fs-long, 50-nJ-energy, 625-nm-wavelength pulses at a 8-kHz repetition rate. The large effective d coefficient results in a relatively high conversion efficiency for such a thin film. A maximum efficiency of 0.13% has been measured. Long-term stable operation was obtained with efficiencies of 0.025% at the damage threshold of approximately 160 GW/cm/sup 2/. The damage threshold is remarkably high for organic materials and was found to be peak intensity rather than average intensity dependent. >

We demonstrate a GaAs/AlGaAs multiple-quantum-well in-line fiber optic intensity modulator. Based on evanescent wave coupling between a GaAs/AlGaAs multiple-quantum-well waveguide and a single-mode fiber, this device concept combines the inherent advantages of in-line fiber devices with high-speed GaAs integrated optoelectronics. The GaAs waveguide uses distributed Bragg mirror layers to phase match to the low-index fiber. Intensity modulation of the transmitted light through the fiber is achieved by tuning the effective propagation index of the GaAs waveguide via the quantum-confined Stark effect. Initial structures show a modulation contrast (ΔT/T) of more than 53%, with an applied voltage of 5 V.

Integrating photodiodes with logic and exploiting on-die interconnect capacitance for energy storage can enable new, low-cost energy harvesting wireless systems. To further explore the tradeoffs between optical efficiency and capacitive energy storage for integrated photodiodes, an array of photovoltaics with various diffractive storage capacitors was designed in TSMC's 90 nm CMOS technology. Transient effects from interfacing the photodiodes with switching regulators were examined. A quantitative comparison between 90 nm and 0.35 μm CMOS logic processes for energy harvesting capabilities was carried out. Measurements show an increase in power generation for the newer CMOS technology, however at the cost of reduced output voltage.

A novel signal processing algorithm for quantifying structural disorder in biological tissue using second harmonic generation (SHG) imaging is described. Both the magnitude and the pattern of disorder in collagenous tissues can be determined with this method. Mathematical models are used to determine the range of disordered states over which the algorithm can be used, because highly disordered biological samples do not generate second harmonic signals. The method is validated by measuring disorder in heated fascicles using SHG and showing that results are significantly correlated with morphometric determination. Applicability of the method to tissue pathology is demonstrated by analysis of a mouse model of intervertebral disk injury. Disks were subjected to tensile or compressive forces in vivo for one week. Structural disorder in the annulus fibrosus was measured by SHG scanning and by standard morphometric analysis. Values for disorder obtained by SHG scanning were significantly correlated with values obtained by morphometry (p

The molecular origins of second order nonlinear effects in collagen fibril assemblies have been identified with sum-frequency generation, infrared and Raman vibrational spectroscopies.

Polysiloxane and conductive polymers were integrated onto a side-polished optical fiber to implement a variable optical attenuator. A conductive polymer thermo-optically altered the refractive index of elastomer placed in close proximity to a fiber core.

Transitions in order/disorder phase states of biological tissues were analyzed using polarization modulated second harmonic generation imaging. The degree of structural disorder was significantly correlated with severity of pathological changes.

The noncentrosymmetric molecular structures responsible for the second order nonlinear effects of collagen fibrils are identified by sum frequency vibrational spectroscopy. CH 2 and C=O functional groups dominate the achiral and chiral spectra, respectively.

An experimental method for quantifying disorder within the anulus fibrosus is described based on polarization-modulated second harmonic generation imaging (PM-SHG-I). This method is demonstrated by imaging the anular lamellar architecture of a mouse model of compressive loading. Results were consistent with those obtained in an earlier study where organization was quantified directed secants image analysis on photomicrographs. In this study the orientation within individual lamellia is quantified by average orientation of the collagen molecules within a defined volume of a single lamellar as measured by the PM-SHG-I. Lamellar boundaries can be identified through the SHG intensity images, and confirmed through co-registration with photomicrographs of the same region. The orientation within the lamellar is quantified by the polarization angle of the maximum second harmonic intensity. PM-SHG-I offers several advantages as compared with the method of directed secants: first, it is nondestructive, allowing repeated measurements of the same tissue; second, images are captured on the order of seconds and capable of obtaining information up to a depth of 200-300 microns, thus allowing for real-time assessment of load damage; third, organization is measured at a much higher resolution, as it is based on disorder within the molecular arrays of a single lamella.

Collagen possesses a strong second order nonlinear susceptibility; when it is irradiated with intense laser light, some of the reflected and transmitted light will have twice the frequency of the incident beam, a phenomenon known as second harmonic generation (SHG). Polarization modulation of an ultra-short pulse laser beam can be used to simultaneously measure collagen fiber orientation, SHG intensity, and a parameter related to the second order non-linear susceptibility. This technique has made it possible to discriminate among patterns of fibrillar orientation in many tissues. In the present study the role that organizational complexity plays in the relationship between nonlinear optical properties and collagen structure is investigated. As a component of tissues and organs, collagen’s structure and function is inextricably intertwined with that of the many other matrix components; to what extent do these noncollagenous components affect its nonlinear properties? To answer this, we investigated SHG in two different collagenous tissues, liver and cartilage; in addition we looked at the effect of progressive pathological changes in these tissues on SHG. At the other end of the spectrum, we studied collagen organized at the minimal level of complexity necessary for SHG detection: fibrils generated from solutions containing only a single type of collagen. Data obtained from these studies suggest that collagen’s strong nonlinear susceptibility, a property no other biologically significant macromolecule shares to the same degree, may serve as more than the basis of a novel imaging device for soft tissue. Collagen’s nonlinear optical properties in conjunction with its vast capacity for self-initiated conformational change--through self-assembly, site recognition, post-translational modification, and the like -make it an attractive candidate molecule for any of several demanding engineering applications, such as nanopatterning.