Modified dual lifetime referencing method for simultaneous optical determination and sensing of two analytes

Simultaneous fluorometric sensing of two analytes becomes possible using a modified dual lifetime referencing (m-DLR) method. In this scheme, two luminescent indicators are needed that have overlapping absorption and emission spectra but largely different decay times. They are excited by a single light source, and both emissions are measured simultaneously. In the frequency domain m-DLR method, the phase of the short-lived fluorescence of a first indicator is referenced against that of the long-lived luminescence of the second indicator. The analytical information is obtained by measurement of the phase shifts at two modulation frequencies. The method is demonstrated to work for the case of dually sensing oxygen and carbon dioxide. It benefits from simple instrumentation and optical setup. The approach is perceived to be of wide applicability. Examples include (1) analysis of two luminescent analytes, (2) analytical determinations that make use of two probes, and (3) sensing of two species such as carbon dioxide and oxygen (as demonstrated here), or oxygen and chlorophyll, provided the luminophores meet the condition of having largely different decay times and overlapping absorption and emission spectra.     

Dual lifetime referencing as applied to a chloride optical sensor

A membrane with an optical response to chloride has been developed that contains two luminophores that display two largely different decay times. The first luminophore (the "reference") is a chloride-insensitive ruthenium metal-ligand complex possessing a decay time in the microsecond range. The second luminophore is the short-lived chloride-quenchable fluorescent probe lucigenin. Both are contained in a hydrogel matrix and are excited by a blue LED emitting sinusoidally modulated light. Under these conditions, the chloride-dependent fluorescence intensity of lucigenin can be converted in an analyte-dependent fluorescence phase shift that depends on the ratio of the two luminescence intensities and can be measured at modulation frequencies of typically 45 kHz. The dynamic range of this sensor can be adjusted by either varying the ratio of the two luminophores or selecting a particular optical filter combination.     

Fluorescence lifetime imaging with near-field scanning optical microscopy

Near-field scanning optical microscopy (NSOM) is a high-resolution scanning probe technique capable of obtaining simultaneous optical and topographic images with spatial resolution of tens of nanometers. We have integrated time-correlated single-photon counting and NSOM to obtain images of fluorescence lifetimes with high spatial resolution. The technique can be used to measure either full fluorescence lifetime decays at individual spots with a spatial resolution of <100 nm or NSOM fluorescence images using fluorescence lifetime as a contrast mechanism. For imaging, a pulsed Ti:sapphire laser was used for sample excitation and fluorescent photons were time correlated and sorted into two time delay bins. The intensity in these bins can be used to estimate the fluorescence lifetime at each pixel in the image. The technique is demonstrated on thin films of poly(9,9'-dioctylfluorene) (PDOF). The fluorescence of PDOF is the results of both inter- and intrapolymer emitting species that can be easily distinguished in the time domain. Fluorescence lifetime imaging with near-field scanning optical microscopy demonstrates how photochemical degradation of the polymer leads to a quenching of short-delay intrachain emission and an increase in the long-delay photons associated with interpolymer emitting species. The images also show how intra- and interpolymer species are uniformly distributed in the films.     

Design and evaluation of a THz time domain imaging system using standard optical design software

A terahertz (THz) time domain imaging system is analyzed and optimized with standard optical design software (ZEMAX). Special requirements to the illumination optics and imaging optics are presented. In the optimized system, off-axis parabolic mirrors and lenses are combined. The system has a numerical aperture of 0.4 and is diffraction limited for field points up to 4 mm and wavelengths down to 750 microm. ZEONEX is used as the lens material. Higher aspherical coefficients are used for correction of spherical aberration and reduction of lens thickness. The lenses were manufactured by ultraprecision machining. For optimization of the system, ray tracing and wave-optical methods were combined. We show how the ZEMAX Gaussian beam analysis tool can be used to evaluate illumination optics. The resolution of the THz system was tested with a wire and a slit target, line gratings of different period, and a Siemens star. The behavior of the temporal line spread function can be modeled with the polychromatic coherent line spread function feature in ZEMAX. The spectral and temporal resolutions of the line gratings are compared with the respective modulation transfer function of ZEMAX. For maximum resolution, the system has to be diffraction limited down to the smallest wavelength of the spectrum of the THz pulse. Then, the resolution on time domain analysis of the pulse maximum can be estimated with the spectral resolution of the center of gravity wavelength. The system resolution near the optical axis on time domain analysis of the pulse maximum is 1 line pair/mm with an intensity contrast of 0.22. The Siemens star is used for estimation of the resolution of the whole system. An eight channel electro-optic sampling system was used for detection. The resolution on time domain analysis of the pulse maximum of all eight channels could be determined with the Siemens star to be 0.7 line pairs/mm.     

Hydrogel-based piezoresistive pH sensors: investigations using FT-IR attenuated total reflection spectroscopic imaging

The strong swelling ability of the pH-responsive poly(acrylic acid)/poly(vinyl alcohol) (PAA/PVA) hydrogel makes the development of a new type of sensor possible, which combines piezoresistive-responsive elements as mechanoelectrical transducers and the phase transition behavior of hydrogels as a chemomechanical transducer. The sensor consists of a pH-responsive PAA/PVA hydrogel and a standard pressure sensor chip. However, a time-dependent sensor output voltage mirrors only the physical swelling process of the hydrogel but not the corresponding chemical reactions. Therefore, an investigation of the swelling behavior of this hydrogel is essential for the optimization of sensor design. In this work, Fourier transform infrared (FT-IR) spectroscopic imaging was used to study the swelling of the hydrogel under in situ conditions. In particular, laterally and time-resolved FT-IR images were obtained in the attenuated total reflection mode and the entire data set of more than 80,000 FT-IR spectra was evaluated by principal component analysis (PCA). The first and third principal components (PCs) indicate the swelling process. Molecular changes within the carboxyl groups were observed in the second and fourth PC and identified as key processes for the swelling behavior. It was found that time-dependent molecular changes are similar to the electrical sensor output signal. The results of the FT-IR spectroscopic images render an improved chemical sensor possible and demonstrate that in situ FT-IR imaging is a powerful method for the characterization of molecular processes within chemical-sensitive materials.     

Fast optical pH manipulation and imaging

We describe a complete system for optical pH manipulation and imaging. The system consists of a photoactive Ruthenium complex capable of inducing a change of more than 5 pH units at the nanosecond time scale. A compatible imaging system acquires microscopic pH images at 1200 fps using a nonexpensive commercial digital camera and an LED illumination system. We use the system as a superb tool to investigate flow in Flow Injection Analysis (FIA) models.     

Fluorescence lifetime standards for time and frequency domain fluorescence spectroscopy

A series of fluorophores with single-exponential fluorescence decays in liquid solution at 20 degrees C were measured independently by nine laboratories using single-photon timing and multifrequency phase and modulation fluorometry instruments with lasers as excitation source. The dyes that can serve as fluorescence lifetime standards for time-domain and frequency-domain measurements are all commercially available, are photostable under the conditions of the measurements, and are soluble in solvents of spectroscopic quality (methanol, cyclohexane, water). These lifetime standards are anthracene, 9-cyanoanthracene, 9,10-diphenylanthracene, N-methylcarbazole, coumarin 153, erythrosin B, N-acetyl-l-tryptophanamide, 1,4-bis(5-phenyloxazol-2-yl)benzene, 2,5-diphenyloxazole, rhodamine B, rubrene, N-(3-sulfopropyl)acridinium, and 1,4-diphenylbenzene. At 20 degrees C, the fluorescence lifetimes vary from 89 ps to 31.2 ns, depending on fluorescent dye and solvent, which is a useful range for modern pico- and nanosecond time-domain or mega- to gigahertz frequency-domain instrumentation. The decay times are independent of the excitation and emission wavelengths. Dependent on the structure of the dye and the solvent, the excitation wavelengths used range from 284 to 575 nm, the emission from 330 to 630 nm. These lifetime standards may be used to either calibrate or test the resolution of time- and frequency-domain instrumentation or as reference compounds to eliminate the color effect in photomultiplier tubes. Statistical analyses by means of two-sample charts indicate that there is no laboratory bias in the lifetime determinations. Moreover, statistical tests show that there is an excellent correlation between the lifetimes estimated by the time-domain and frequency-domain fluorometries. Comprehensive tables compiling the results for 20 (fluorescence lifetime standard/solvent) combinations are given.     

Segmented multimode to dual port slot couplers as compact optical sensors with improved sensitivity

A segmented silicon based multimode to dual port slot structure on silicon-on-insulator platform is proposed which can be used as a refractive-index sensing device. The introduction of segmentation leads to tuning the effective index of the device which results in increasing compactness of the sensing device. Although the structure supporting TM mode is more compact than TE mode, but TE mode is considered here as vertical slots in the output section enhances optical signal in the slots for TE mode only. By considering dual output, the device length is reduced further as dual self-imaging length is less compared to single self-imaging distance for symmetrical multimode section input. The surface sensitivity of the structure has a typical value of～2249?nm/RIU. Relative sensitivity can be calculated from the ratio of field amplitudes of the arms of the dual output. Matrix method and 2D FDTD is used for the entire analysis.?     

Integrated semiconductor optical sensors for cellular and neural imaging

We review integrated optical sensors for functional brain imaging, localized index-of-refraction sensing as part of a lab-on-a-chip, and in vivo continuous monitoring of tumor and cancer stem cells. We present semiconductor-based sensors and imaging systems for these applications. Measured intrinsic optical signals and tissue optics simulations indicate the need for high dynamic range and low dark-current neural sensors. Simulated and measured reflectance spectra from our guided resonance filter demonstrate the capability for index-of-refraction sensing on cellular scales, compatible with integrated biosensors. Finally, we characterized a thermally evaporated emission filter that can be used to improve sensitivity for in vivo fluorescence sensing.     

A design method of spatiotemporal optical pulse using level-set based time domain topology optimization

Recently, two emerging areas of photonics research, ultrafast photonics, and nanophotonics have started to come together. One of the main problems in this field is the precise control of spatial and temporal profiles of the optical pulses. In this paper, we propose a design method for user-specified spatiotemporal optical pulses using a level set-based time-domain topology optimization method. In the proposed method, the optimization problem is formulated based on time domain Maxwell equations so that the spatiotemporal optical pulses can be treated directly. The objective function is defined using the envelope information of the pulses, and an efficient way to calculate this information, based on calculations of the complex electromagnetic field, is introduced. A level set-based topology optimization method is applied to obtain optimized configurations. Using the proposed method, the spatiotemporal user-specified pulse profiles can be designed by modifying the structural details of the nanostructures through which the pulses propagate. As a simple example, we demonstrate that the optimized structures focus optical pulses into a single or multiple focal points with a user-specified pulse-width. The results show that the proposed method is able to design highly controlled spatiotemporal optical pulses by engineering the nanophotonic structure.     

Fluorescence lifetime imaging by using time-gated data acquisition

The use of the time gating technique for lifetime reconstruction in the Fourier domain is a novel technique. Time gating provides sufficient data points in the time domain for reliable application of the Fourier transform, which is essential for the time deconvolution of the system of the integral equations employed in the reconstruction. The Fourier domain telegraph equation is employed to model the light transport, which allows a sufficiently broad interval of frequencies to be covered. Reconstructed images contain enough information needed for recovering the lifetime distribution in a sample for any given frequency within the megahertz-gigahertz band. The use of this technique is essential for recovering time-dependent information in fluorescence imaging. This technique was applied in reconstruction of the lifetime distribution of four tubes filled with Rhodamine 6G embedded inside a highly scattering slab. Relatively accurate fluorescence lifetime reconstruction demonstrates the effectiveness and the potential of the proposed technique.     

Wide-field fluorescence lifetime imaging with optical sectioning and spectral resolution applied to biological samples

Abstract. Wide-field fluorescence lifetime imaging with spectral resolution and optical sectioning has been performed to achieve five-dimensional fluorescence microscopy. Spectral filtering has been shown to have the potential to provide functional information about biological tissue by simultaneously measuring the spectral/lifetime signature of the sample. The potential to use multispectral imaging to separate cellular components spatially by their different emission wavelengths has also been demonstrated thus reducing artefacts in the calculated lifetime maps. The instrument is based on diode-pumped solid-state laser technology and an ultrafast gated optical image intensifier. Also reported is the use of a picosecond blue laser diode as the excitation source to produce a fluorescence lifetime microscope with a footprint of less than 0.25m².     

Sidelobe suppression in ultrasound imaging using dual apodization with cross-correlation

This paper introduces a novel sidelobe and clutter suppression method in ultrasound imaging called dual apodization with cross-correlation or DAX. DAX dramatically improves the contrast-to-noise ratio (CNR) allowing for easier visualization of anechoic cysts and blood vessels. This technique uses dual apodization or weighting strategies that are effective in removing or minimizing clutter and efficient in terms of computational load and hardware/software needs. This dual apodization allows us to determine the amount of mainlobe versus clutter contribution in a signal by cross-correlating RF data acquired from 2 apodization functions. Simulation results using a 128 element 5 MHz linear array show an improvement in CNR of 139% compared with standard beamformed data with uniform apodization in a 3 mm diameter anechoic cylindrical cyst. Experimental CNR using a tissue-mimicking phantom with the same sized cyst shows an improvement of 123% in a DAX processed image.     

Thin-film optical sensors with silicon-compatible materials

Antireflection filters based on multilayer stacks of dielectric and polysilicon films on monocrystalline silicon combined with charge collection in different (poly)Si layers can be used to realize sensors with a programmable spectral response controlled by weighted summing of the photocurrents detected in the polysilicon and the substrate. Thus, employing both interference and selective absorption of light yields increased photoelectric efficiency and improved flexibility of spectral control and enables on-chip integration of the detector(s) with the signal conditioning and processing circuits. The potential of thin-film color sensors has been evaluated for this purpose. However, for practical implementation of such structures the problems associated with the realization of reliable photodetectors in polysilicon must also be considered. Phosphorus passivation of the grain-boundary states has been employed to yield polysilicon photodiodes with improved electrical characteristics and reliable light and color detection. We present the design methods of thin-film color sensors employing silicon-compatible materials only. The measurement results of a fabricated structure fully demonstrate that such sensors can be realized with good spectral selectivity.     

海洋涡旋地震成像的时域有限差分仿真

传统的水声建模方法只考虑水中声波的折射影响,忽略了水层间的声反射,因此不能对反射波地震勘探的海水分层结构成像结果进行分析。文章首先建立了基于时域有限差分的低频宽带脉冲信号传输预报模型,该模型考虑海水分层结构的声反射影响,通过数值求解波动方程计算低频宽带脉冲信号近距离传播过程中的时域波形;其次将某次海上多道地震测量的水文条件作为输入环境参数,计算了近海面地震勘探接收阵所接收的气枪低频脉冲信号时域波形;最后按照反射波地震勘探处理流程,对接收阵信号在共反射点进行相干累加和地震成像,所得到的海水分层结构成像结果与海上多道测量成像结果符合较好。仿真结果验证了水层声反射对海水分层结构反射波地震成像的影响。     

Surface imaging using holographic optical tweezers

We present an imaging technique using an optically trapped cigar-shaped probe controlled using holographic optical tweezers. The probe is raster scanned over a surface, allowing an image to be taken in a manner analogous to scanning probe microscopy (SPM), with automatic closed loop feedback control provided by analysis of the probe position recorded using a high speed CMOS camera. The probe is held using two optical traps centred at least 10 μm from the ends, minimizing laser illumination of the tip, so reducing the chance of optical damage to delicate samples. The technique imparts less force on samples than contact SPM techniques, and allows highly curved and strongly scattering samples to be imaged, which present difficulties for imaging using photonic force microscopy. To calibrate our technique, we first image a known sample--the interface between two 8 μm polystyrene beads. We then demonstrate the advantages of this technique by imaging the surface of the soft alga Pseudopediastrum. The scattering force of our laser applied directly onto this sample is enough to remove it from the surface, but we can use our technique to image the algal surface with minimal disruption while it is alive, not adhered and in physiological conditions. The resolution is currently equivalent to confocal microscopy, but as our technique is not diffraction limited, there is scope for significant improvement by reducing the tip diameter and limiting the thermal motion of the probe.     

An optical fiber sensor for remote pH sensing and imaging

A fiber-optical probe for pH sensing and real-time imaging is successfully fabricated by connecting a polymer imaging fiber and a gradient index (GRIN) lens rod which was modified with a sensing film. By employing an improved metallographic microscope, an optical system is designed to cooperate with the probe. This novel technique has high-quality imaging capabilities for observing remote samples while measuring pH. The linear range of the probe is pH 1.2-3.5. This technique overcomes the difficulty that high-quality images cannot be obtained when directly using conventional imaging bundles for pH sensing and imaging. As preliminary applications, the corrosion behavior of an iron screw and the reaction process of rust were investigated in buffer solutions of pH 2.0 and 2.9, respectively. The experiment demonstrated that the pH values of the analytes' surface were higher than that of buffer solutions due to the chemical reaction. It provides great potential for applications in optical multifunctional detection, especially in chemical sensing and biosensing.     

Fluorescent ion-selective nanosensors for intracellular analysis with improved lifetime and size

We describe the synthesis and characterization of sodium-selective polymeric nanosensors that improves upon the lifetime and size of previous fiberless nanosensors. Sonication is used to form the polymer nanospheres that contain all the components needed for ion sensing. Even though the size is small (approximately 120 nm), the lifetime of these sensors in solution is on the order of a week. The surface coating has also been optimized for stability, biocompatibility, and ease of chemical modification.     

Strain monitoring in composite-strengthened concrete structures using optical fibre sensors

In this paper, the mechanical behaviour of the composite-strengthened concrete structures is addressed. Optical fibre sensor presents a great deal of potential in monitoring the structural health condition of civil infrastructure elements after strengthening by externally bonded composite materials. The use of embedded optical fibre sensor for strain and temperature monitoring enables to reveal the status of the composite-strengthened structure in real-time remotely. In this paper, an experimental investigation on the composite-strengthened concrete structures with the embedment of fibre-optic Bragg grating (FBG) sensors is presented. Single- and multiplexed-point strain measuring techniques were used to measure strains of the structures. Frequency modulated continuous wave (FMCW) method was used to measure strains in different points of the structure with using only one single optical fibre. All strains measured from the sensors were compared to conventional surface mounted strain gauges. Experimental results show that the use of the embedded FBG sensor can measure strain accurately and provide information to the operator that the structure is subjected to debond or micro-crack failure. Multiplexed FBG strain sensors enable to measure strain in different locations by occupying only one tiny optical fibre. Reduction of strength in composite laminate is resulted if the embedded optical fibre is aligned perpendicular to the load-bearing direction of the structure.