Clinical imaging fluorescence apparatus for the endoscopic photodetection of early cancers by use of Photofrin II

A fluorescence imaging device applied to the detection of early cancer is described. The apparatus is based on the imaging of laser-induced fluorescence of a dye that localizes in a tumor with a higher concentration than in the surrounding normal tissue after iv injection. Tests carried out in the upper aerodigestive tract, the tracheobronchial tree, and the esophagus with Photofrin II (1 mg/kg of body weight) as the fluorescent agent are reported as examples. The fluorescence is induced by violet (410-nm) light from a continuous-wave (cw) krypton-ion laser. The fluorescence contrast between tumor and surrounding tissue is enhanced by real-time image processing. This is done by the simultaneous recording of the fluorescence image in two spectral domains (470-600 and 600-720 nm), after which these two images are digitized and manipulated with a mathematical operator (look-up table) at video frequency. Among the 7 photodetections performed in the tracheobronchial tree, 6 were successful, whereas it was the case for only 5 of the 15 lesions investigated in squamous mucosa (upper aerodigestive tract and esophagus). The sources of false positives and false negatives are evaluated in terms of the fluorescent dye, tissue optical properties, and illumination optics.     

Influence of squeezing bandwidths on resonance fluorescence

Abstract

We present an analytical technique for investigating the behaviour of a coherently driven atom damped by a squeezed vacuum with finite squeezing bandwidth. A master equation and analytic expressions for the fluorescent spectrum are derived for the simple case of a two-level atom exactly resonant with the frequencies of both the squeezed field and the driving field. Suitable choices of driving strength, phase and squeezing source lead to either sub-natural lincwidths of the Rabi sidebands or a sub-natural linewidth of the central spectral peak or indeed sub-natural linewidths of all three spectral peaks when squeezing bandwidths are taken into account. We also establish the remarkable result that in the strong driving regime, the width of the central peak of the fluorescent spectrum depends solely on the squeezing found at the Rabi sideband frequencies.     

Ultrabroadband photodetection based on graphene ink

We report photodetection in a very large spectral bandwidth, which encompasses ultraviolet, visible and near infrared, using graphene inks or graphene inks functionalized with either gold or silver nanoparticles, or gold nanoparticles further encapsulated with bovine serum albumin deposited on interdigitated electrodes fabricated on a silicon dioxide/silicon substrate. In contrast to gold-functionalized graphene inks, which have responsivities better than 1 mA W(-1) at a 0.1 V bias over the huge bandwidth extending from 215 to 2500 nm, Ag-functionalized inks show at least a four-fold increased responsivity, with a record value of 13.7 mA W(-1) in near infrared.     

Room-temperature photodetection dynamics of single GaN nanowires

We report on the photocurrent behavior of single GaN n-i-n nanowires (NWs) grown by plasma-assisted molecular-beam epitaxy on Si(111). These structures present a photoconductive gain in the range of 10(5)-10(8) and an ultraviolet (350 nm) to visible (450 nm) responsivity ratio larger than 6 orders of magnitude. Polarized light couples with the NW geometry with a maximum photoresponse for polarization along the NW axis. The photocurrent scales sublinearly with optical power, following a I ~ P(β) law (β < 1) in the measured range with β increasing with the measuring frequency. The photocurrent time response remains in the millisecond range, which is in contrast to the persistent (hours) photoconductivity effects observed in two-dimensional photoconductors. The photocurrent is independent of the measuring atmosphere, either in the air or in vacuum. Results are interpreted taking into account the effect of surface states and the total depletion of the NW intrinsic region.     

The asymptotic behaviour of the spectrum of resonance fluorescence

Abstract

The spectrum of resonance fluorescence from laser-driven multi-level systems is investigated and especially its asymptotic frequency dependence in the wings. Analytical results are presented and the transitions within multilevel systems are classified into three categories according to the asymptotic frequency dependence of their spectra. Under certain conditions an arbitrarily fast asymptotic fall-off behaviour of the spectrum is found which, for intermediately strong driving fields, leads to subnatural linewidths. Numerical examples are presented to illustrate the analytical results.     

Novel superconducting proximized heterostructures for ultrafast photodetection

Weak ferromagnet/superconductor (F/S) proximity bilayers have been fabricated and characterized for basic physics studies concerning the ultrafast carrier dynamics in layered materials. The normalized reflectivity change (ΔR/R) as a function of the time delay between the pump and the probe laser beams has been measured in F/S heterostructures formed by a low critical temperature superconductor (Nb) with a NiCu overlayer, and a high T_c superconductor (YBCO) covered by Au/NiCu and manganite (LSMO) overlayers. The attention is mainly focused to the investigation of nonequilibrium excitation dynamics inside different bilayers in the low temperature region. The presence of the weak ferromagnetic overlayer produces faster optical relaxation times with respect to sole superconducting films. The results are promising in view of potential applications as ultrafast kinetic inductance superconducting photodetectors as confirmed by preliminary time-resolved photoimpedance experiments on both Nb and NiCu(21 nm)/Nb(70 nm) samples.     

Fluorescence coincidence spectroscopy for single-molecule fluorescence resonance energy-transfer measurements

Single-molecule fluorescence resonance energy transfer (FRET) is commonly used to probe different conformations and conformational dynamics of single biomolecules. However, the analysis of raw burst traces is not always straightforward. The presence of a "zero peak" and the skewness of peaks at high and low FRET efficiencies in proximity ratio histograms make the accurate evaluation of the histogram a challenging task. This is further compounded by the difficulty associated with siting two fluorophores in optimal range of each other. Here we present an alternative method of analysis, based on handling coincident FRET photon bursts, that addresses these problems. In addition, we demonstrate methods to enhance coincidence levels and thus the accuracy of FRET determination: the use of dual-color excitation, including direct excitation of the acceptor fluorophore; the addition of a remote dye to the biomolecule, not involved in the FRET process; or a combination of the two. We show the advantages of dual excitation by studying several labeled double-stranded DNA samples as FRET models. This method extends the application of single-molecule FRET to more complicated biological systems where only a small fraction of complexes are fully assembled.     

Plasmon-enhanced resonance Raman scattering and fluorescence in Langmuir-Blodgett monolayers

Localized surface plasmon resonances in silver and gold nanostructures are engaged to enhance the inelastic Raman scattering and the fluorescence of a phopholipid containing a sulforhodamine 101 acid chloride dye known as Texas Red. The most efficient coupling and enhancement are attained when the excitation laser line is in resonance with both the chromophore and the plasmon absorption of the nanostructure, the case of surface-enhanced resonance Raman scattering, allowing single-molecule detection. The tagged phospholipid was incorporated into a single fatty acid Langmuir monolayer at varying concentrations and transferred onto an evaporated Ag nanoparticle film. Surface-enhanced fluorescence is achieved using shell-isolated silica-coated gold nanoparticles, an emission enhancement named SHINEF.     

Array of ZnO nanoparticle-sensitized ZnO nanorods for UV photodetection

Vertically aligned ZnO nanorod (NR) arrays have been successfully synthesized on ITO-glass substrate by hydrothermal growth. Chemical bath deposition method was used to deposit ZnO nanoparticles (NPs) onto the ZnO NRs. These structures were applied in fabricating ZnO NPs sensitized ultraviolet (UV) photodetectors (PDs). Incorporation of ZnO NPs onto ZnO NRs results in distinct improvement of optical properties of ZnO NRs, i.e., significant enhancement of emission as well as effective suppression of defects emission in ZnO. Furthermore, there is a noticeable blue-shift in absorption spectra compared with that of ZnO NRs structure. I–V characteristics show that the sensitized structure improved photocurrent almost twice that of unsensitized ZnO NRs. Consequently, these findings may open new opportunities for the integration of different ZnO nanostructures for application in UV region particularly fabrication of UV PDs.     

Transport and photodetection in self-assembled semiconductor quantum dots

A great step forward in science and technology was made when it was discovered that lattice mismatch can be used to grow highly ordered, artificial atom-like structures called self-assembled quantum dots. Several groups have in the meantime successfully demonstrated useful infrared photodetection devices which are based on this technology. The new physics is fascinating, and there is no doubt that many new applications will be found when we have developed a better understanding of the underlying physical processes, and in particular when we have learned how to integrate the exciting new developments made in nanoscopic addressing and molecular self-assembly methods with semiconducting dots. In this paper we examine the scientific and technical questions encountered in current state of the art infrared detector technology and suggest ways of overcoming these difficulties. Promoting simple physical pictures, we focus in particular on the problem of high temperature detector operation and discuss the origin of dark current, noise, and photoresponse.     

Homogeneous immunoassay based on two-photon excitation fluorescence resonance energy transfer

A two-photon excitable small organic molecule (abbreviated as TP-NH 2) with large two-photon absorption cross section and competitive fluorescence quantum yield was prepared, which emitted fluorescence in the visible region upon excitation at 800 nm. Using the TP-NH 2 molecule as an energy donor, a two-photon excitation fluorescence resonance energy-transfer (TPE-FRET) based homogeneous immunoassay method was proposed. The donor and the acceptor (DABS-Cl, a dark quencher) were labeled to bovine serum albumin (BSA) separately, and anti-BSA protein was determined by employing an antibody bridging assay scheme. Rabbit anti-BSA serum containing other biomolecules was intentionally used as the sample to introduce interference. A parallel assay was performed using the traditional one-photon excitation FRET model, which failed to carry out quantitative determination due to the serious background luminescence arising from those biomolecules in the sample. The TPE-FRET model showed its strong ability to overcome the problem of autofluorescence and provided satisfying analytical performance. Quite good sensitivity and wide linear range (0.05-2.5 nM) for anti-BSA protein was obtained. The results of this work suggest that TPE-FRET could be a promising technique for homogeneous assays excluding separation steps, especially in complicated biological sample matrixes.     

Tomographic imaging of ratiometric fluorescence resonance energy transfer in scattering media

A method to visualize and quantify fluorescence resonance energy transfer (FRET) in scattering media is proposed. It combines the ratiometric FRET method with fluorescence molecular tomography (FMT) in continuous wave (CW) mode. To evaluate the performance of the proposed method, experiments on a tissue-mimicking phantom are carried out. The results demonstrate that the proposed approach is capable of visualizing and quantifying the FRET distribution in scattering media, which implies the further application of the ratiometric assay in in vivo studies.     

Dendrimer-based nanoprobe for dual modality magnetic resonance and fluorescence imaging

A novel PAMAM dendrimer-based nanoprobe for dual magnetic resonance and fluorescence imaging modalities was synthesized. Fluorescence studies revealed that Gd(III) complexation to the probe has no effect on the quantum yield; however, increases in the dye content resulted in partial quenching. The potential of the new nanoprobe, G6-(Cy5.5)(1.25)(1B4M-Gd)(145), as a dual modality imaging agent was demonstrated in vivo by the efficient visualization of sentinel lymph nodes in mice by both MRI and fluorescence imaging modalities.     

Spectroscopic features of dual fluorescence/luminescence resonance energy-transfer molecular beacons

Molecular beacons have the potential to become a powerful tool in gene detection and quantification in living cells. Here we report a novel dual molecular beacons approach to reduce false-positive signals in detecting target nucleic acids in homogeneous assays. A pair of molecular beacons, each containing a fluorescence quencher and a reporter fluorophore, one with a donor and a second with an acceptor fluorophore, hybridize to adjacent regions on the same target resulting in fluorescence resonance energy transfer (FRET). The detection of a FRET signal leads to a substantially increased signal-to-background ratio compared with that seen in single molecular beacon assays and enables discrimination between fluorescence due to specific probe/target hybridization and a variety of possible false-positive events. Further, when a lanthanide chelate is used as a donor in a dual-probe assay, extremely high signal-to-background ratios can be achieved owing to the long lifetime and sharp emission peaks of the donor and the time-gated detection of acceptor fluorescence emission. These new approaches allow for the ultrasensitive detection of target molecules in a way that could be readily applied to real-time imaging of gene expression in living cells.     

Continuous monitoring of adriamycin in vivo using fiber optic-based fluorescence chemical sensor

A novel method using a fiber optic-based fluorescence chemical sensor (FOCS) was developed for the preparation of on-line continuous monitoring of a drug in animals. First, an accurate optical design was used to enhance the intensity of light from a 100-microm optic fiber so the fluorescence signal can be detected. Second, A new sol-gel method was used to fix the fluorescence substance 4-(N,N-dioctyl)amino-7-nitrobenz-2-oxa-1,3-diazole (D-70) on the tip of the fiber. The variation in the quenching means variation in the concentration of Adriamycin (ADM) in rabbit's blood. ADM is determined by FOCS based on fluorescence multiple quenching. In a simple animal model, the carotid artery was catheterized with a cannula, housing a 100-microm optic fiber. The average recovery of all the tested compounds within the set concentration range was 99.4-106.2%; the within-day and between-day repeatability values were acceptable between 6.6-11.4% and 5.9-11.7%. The method permitted detection limits as low as 0.057 microg x mL(-1) at a signal-to-noise ratio of 3. The fiber optic chemical sensor is potentially useful for monitoring blood concentrations of drugs and metabilities in the biomedical field.     

Measuring the absolute photodetection efficiency using photon number correlations

We present two methods for determining the absolute detection efficiency of photon-counting detectors directly from their singles rates under illumination from a nonclassical light source. One method is based on a continuous variable analog to coincidence counting in discrete photon experiments, but it does not actually rely on high detector time resolutions. The second method is based on difference detection, which is a typical detection scheme in continuous variable quantum optics experiments. Since no coincidence detection is required with either method, they are useful for detection efficiency measurements of photodetectors with detector time resolutions far too low to resolve coincidence events.     

Quantum dot-fluorescent protein pairs as novel fluorescence resonance energy transfer probes

Fluorescence resonance energy transfer (FRET) characteristics, including the efficiency, donor-acceptor distance, and binding strength of six fluorescent protein (FP)-quantum dot (QD) pairs were quantified, demonstrating that FPs are efficient acceptors for QD donors with up to 90% quenching of QD fluorescence and that polyhistidine coordination to QD core-shell surface is a straightforward and effective means of conjugating proteins to commercially available QDs. This provides a novel approach to developing QD-based FRET probes for biomedical applications.