Fluorescent Dye Solution Research Articles

UV/Visible Diffusion-Ordered Spectroscopy: A Simultaneous Probe of Molecular Size and Electronic Absorption.

Based on concepts from nuclear magnetic resonance, we have developed UV/vis diffusion-ordered spectroscopy, which simultaneously probes the size and electronic absorption spectrum of molecules and particles. We use simple flow technology to create a step-function concentration profile inside an optical sample cell, and by measuring the time-dependent absorption spectrum in an initially solvent-filled part of the sample volume, we obtain the diffusion coefficients and UV/vis spectra of the species present in the sample solution. From these data, we construct a two-dimensional spectrum with absorption wavelength on one axis and diffusion coefficient (or equivalently, size) on the other, in which the UV/vis spectrum of a mixture with different molecular sizes is separated into the spectra of the different species, sorted by size. We demonstrate this method on mixed solutions of fluorescent dyes, biomolecules, and the UV-absorbing components of coffee, caffeine, and chlorogenic acid, all with concentrations in the μM range.

On-Target Deposition from Two Engine-Powered Sprayers in Medium-Foliage-Density Citrus Canopies

Spray penetration into citrus canopies is critical for adequate coverage and deposition to ensure effective pest control. However, mismatch of air assistance to target canopy characteristics can lead to unintended spraying losses through overpenetration. To evaluate the effect of air assistance on on-target deposition, two sprayers (surrogates for airflow rates) were used to apply a fluorescent tracer dye solution @ a target concentration of 300 ppm to 16 medium-foliage-density tree blocks in a commercial mandarin orchard. The complete factorial experiment in three replications, also designed for validating a model-based spray decision support tool, comprised two forward travel speeds (1.6 and 4.8 km/h), two disc-core nozzles (TeeJet® D3-25 and D6-45), and either one or two nozzle rows to obtain a wide range of application rates (496 to 9719 L/ha). Dye deposition significantly decreased with canopy depth (p =< 0.001) by nearly seven times across the 3.4 m wide canopies but was not significant over 1.2 to 2.2 m sampling height (p = 0.867). Deposition obtained with the low-airflow-rate sprayer was significantly greater (p =< 0.001) than that obtained with the high-airflow-rate sprayer over the dose range likely due to too much air pushing out spray droplets. This study underscores the importance of matching the air assistance of orchard sprayers to the target canopy.

Concentration polarization around polyelectrolyte-coated electrodes. Model and observations

An investigation on the phenomenon of concentration polarization (CP) in conducting porous particles is presented in this work, considering both bare and polyelectrolyte-coated particles. The conducting nature of the porous structure brings about the induction of a surface (and hence volume) charge distribution by the applied external field. The polymer charge (with its counterions from solution) is superimposed on this field-induced component. From the solution of Poisson’s equation, the concentration and potential profiles are evaluated, and from them, the concentration polarization can be calculated. The results are presented as concentration perturbation as a function of time after application of the field, both for bare and coated particles. Experiments are also performed aimed at measuring the CP using a solution of fluorescent dye (rhodamine B). From the increase or decrease of fluorescence, the concentration perturbations are observed around the particle. Importantly, depletion of concentration is observed on both sides of the particle when this is bare. In contrast, if the particles are coated, the classical pattern of a pole of increased concentration and an opposite one of decreased concentration is found. Dielectric dispersion experiments in suspensions of bare and brush-coated particles confirm this fact.

Comparison of water-sensitive paper, Kromekote and Mylar collectors for droplet deposition with a visible fluorescent dye solution

The study was conducted at the University of Nebraska Pesticide Application and Technology Laboratory in North Platte, Nebraska in July 2015. Two application volume rates (100 and 200 l · ha−) and three nozzle types (XR, AIXR, TTI) were selected at two flow rates (0.8 and 1.6 l · min−) and at a single application speed of 7.7 km · h−. Each collector type [Mylar washed (MW), Mylar image analysis (MIA), water-sensitive paper (WSP), and Kromekote (KK)] was arranged in a randomized complete block design. Each nozzle treatment was replicated twice, providing six cards of each collector type for each nozzle treatment. A water + 0.4% v/v Rhodamine WT spray solution was applied, given the fluorescent and visible qualities of Rhodamine, which allows it to be applied over all the collector types. MW had the highest coverage at 18.3% across nozzle type, followed by WSP at 18%, KK at 12% and lastly by MIA at 4%. MW resulted in a 58% increase in coverage, WSP in a 56% increase, and KK only an increase of 39% when the volume rate was doubled from 100 l · ha− to 200 l · ha− across nozzle type. MW coverage was similar to KK for half of the nozzles (XR 11002, XR 11004, AIXR 11002). Droplet number density fixed effects were all significant for nozzle type and collector type (p < 0.001) as was the interaction of nozzle type and collector type (p < 0.001). Results from this study suggest a strong correlation to data produced with WSP and MW collectors, as there was full agreement between both types except for the TTI 11004. Using both collector types in the same study would allow for a visual understanding of the distribution of the spray, while also giving an idea of the concentration of that distribution.

Ultrafast pulse duration measurement method of near-infrared pulses for a broad range of wavelengths using two-photon absorption in a liquid and fluorescent dye solution.

A novel characterization method to measure the pulse duration of ultrafast near-IR pulses is introduced, which uses simple tabletop optics, is relatively inexpensive, and is expected to work in a broad wavelength range. Our diagnostic tool quantitatively characterizes the laser pulse duration of any near-IR wavelength assuming a Gaussian pulse shape with a linear chirp. We negatively prechirp near-IR pulses with a home-built broadband pulse compressor (BPC) and send this prechirped beam through a cell filled with a low-molar solution of a fluorescent dye in a liquid. After two-photon absorption, this dye fluoresces in the visible, and we record this visible signal as a function of the propagation distance in the liquid cell. We calibrate the group velocity dispersion (GVD) of our home-built BPC device against the known GVD of the compressor of our 800nm laser and confirm this value using geometric considerations. Now knowing the GVD of BPC and the recorded visible signal for various amounts of negative chirp, let us extract the smallest pulse duration of the near-IR pulse from this visible signal. As a useful corollary, our analysis also enables the direct measurement of the GVD for liquids and the indirect measurement of the absorption coefficient for liquids in the near-IR range, in contrast to indirect GVD measurements that rely on methods such as the double derivative of the refractive index.

Fluorescence Excitation-Emission-Matrix Imaging.

We present a 4-dimensional (4D) fluorescence imaging system in which each of the 65,536 pixels in the image array contains an excitation-emission-matrix spectrum with 31 excitation wavelengths and 8 emission wavelengths (x, y, λexc, λem). Hadamard-transform multiplexing of the excitation light from a 31-channel programmable light source allows for an increase in the data acquisition rate so that each 65,536-pixel image can be obtained within 8 s. The system is demonstrated and characterized using, first, a 4D image of 10 capillaries filled with four dye solutions and their binary and ternary mixtures, and, second, using a sequence of about 100 images of layered fluorescent dye solutions and their changing fluorescence as a function of temperature. Multivariate analysis using parallel factor analysis produces images of the spatial distribution of the fluorophores together with their relative intensity as a function of time.

Impact of Immediate and Delayed Photo-activation of Self-adhesive Resin Cements on Bonding Efficacy and Water Uptake Under Simulated Pulpal Pressure.

This study investigated the effect of immediate versus delayed photo-activation on the bonding performance and water uptake of self-adhesive (SA) resin cements under simulated pulpal pressure (SPP). The occlusal dentin surface was exposed in 66 extracted third molars. Resin composite cylinders were cemented to dentin under SPP, with either RelyX Unicem 2 (RU) (3M Oral Care, St Paul, MN, USA) or Maxcem Elite (MC) (Kerr, Orange, CA, USA). Each cement group was equally divided into three groups (n=8 each) according to the time elapsed between placement and photo-activation: immediate activation (IM), 30-second delayed activation (D30), or 120-second delayed activation (D120). Shear bond strength (SBS) was measured, and the type of failure was determined using a stereomicroscope. Three additional samples from each experimental subgroup were used for confocal laser scanning microscopy (CLSM) analysis. A fluorescent dye solution was added to the pulpal fluid reservoir, then a CLSM was used to detect the dye distribution within the tooth-restoration interface. Two-way analysis of variance (ANOVA) and the Tukey post-hoc test were used to analyze the SBS results (α=0.05). D30 resulted in a significantly higher mean SBS in the two cement groups than IM and D120 (p<0.05). RU showed significantly higher SBS values than MC regardless of the time of light activation (p<0.05). RU showed less dye uptake confined to the cement-dentin interface compared to the MC groups, which showed dye uptake throughout the entire thickness of the cement layer and gap formation at the interface, especially in the D120 group. The 30-second photo-activation delay group significantly improved the bond strength of SA cements. Delaying the photo-activation to 120 seconds increased pulpal fluid uptake by SA cements and compromised the integrity of the bonded interfaces.

Non-Destructive Measurement of Acetic Acid and Its Distribution in a Photovoltaic Module during Damp Heat Testing Using pH-Sensitive Fluorescent Dye Sensors

An optical pH sensor that enables the non-destructive measurement of acetic acid and its distribution in a photovoltaic module during damp heat (DH) testing is reported. The sensor was fabricated by impregnating a solution of a pH-sensitive fluorescent dye into a fluororesin membrane filter, which was then dried. While conducting the DH test, fluorescence spectra from 20 pH sensors were periodically recorded and converted into pH values using a predetermined calibration curve. As a result, we succeeded in measuring changes in pH with a DH test time of up to 2000 h, and it was possible to obtain information on the pH distribution in the module. We also confirmed no change in pH in a module with a silicone encapsulant free from acetic acid, and revealed that the sensor that we developed does not respond to moisture and heat, but only to acetic acid.

Mixing enhancement in T-junction microchannel with acoustic streaming induced by triangular structure.

The T-shaped microchannel system is used to mix similar or different fluids, and the laminar flow nature makes the mixing at the entrance junction region a challenging task. Acoustic streaming is a steady vortical flow phenomenon that can be produced in the microchannel by oscillating acoustic transducer around the sharp edge tip structure. In this study, the acoustic streaming is produced using a triangular structure with tip angles of 22.62°, 33.4°, and 61.91°, which is placed at the entrance junction region and mixes the inlets flow from two directions. The acoustic streaming flow patterns were investigated using micro-particle image velocimetry (μPIV) in various tip edge angles, flow rate, oscillation frequency, and amplitude. The velocity and vorticity profiles show that a pair of counter-rotating streaming vortices were created around the sharp triangle structure and raised the Z vorticity up to 10 times more than the case without acoustic streaming. The mixing experiments were performed by using fluorescent green dye solution and de-ionized water and evaluated its performance with the degree of mixing (M) at different amplitudes, flow rates, frequencies, and tip edge angles using the grayscale value of pixel intensity. The degree of mixing characterized was found significantly improved to 0.769 with acoustic streaming from 0.4017 without acoustic streaming, in the case of 0.008 μl/min flow rate and 38 V oscillation amplitude at y = 2.15 mm. The results suggested that the creation of acoustic streaming around the entrance junction region promotes the mixing of two fluids inside the microchannel, which is restricted by the laminar flow conditions.

Locally controlled two-photon excited fluorescence by correlated ultrafast intensity fluctuations

We propose a method for improving the imaging depth of two-photon excitation microscopy using correlated ultrafast intensity fluctuations within pulses. As a proof of principle, we experimentally demonstrate local control of two-photon excitation by using the ultrafast intensity cross-correlation generated by high-gain parametric down-conversion. We show that only the fluorescence intensity emitted from deep inside the fluorescent dye solution can be modulated by harnessing the correlation at ultrashort time scales. It is expected that the influence of the background photons can be suppressed by applying this technique to the two-photon excitation microscopy.

Microplastics in the snow cover of the south of Western Siberia

Plastics production is growing all over the world, but the total emission of microplastics is unfortunately not reducing. In the external environment, plastics degrade over time. This leads to the appearance of smaller particles or fibers which are microplastics at sizes of less than 5 mm. Atmospheric transfer is for microplastics one of the main ways of entering the environment. The purpose of this study is to identify microplastics in the snow cover of the south of Western Siberia, as well as preliminarily assess the possible sources of their supply. Snow samples are taken during a period of maximum snow accumulation (March, 2020) according to requirements developed by the Norwegian Institute for Air Research. The microparticles are visually evaluated using a digital microscope, Dino-Lite AM211, a fluorescent dye solution, and a Wood lamp. To assess the possible atmospheric distribution and deposition of the particles, an analysis of the synoptic conditions based on data of NCEP/NCAR, ERA5, and HYSPLIT is performed. The microscopic analysis has shown that microplastics (namely, films, fibers, and granules) have been detected in 16 of the 18 snow samples in the city of Barnaul and adjacent territories, as well as fibers and granules have been revealed in 3 snow samples from the Kasmala River basin.

The Effects of Cyclic Loading and Motion on the Implant-Cement Interface and Cement Mantle of PEEK and Cobalt-Chromium Femoral Total Knee Arthroplasty Implants: A Preliminary Study.

This study investigated the fixation of a cemented PEEK femoral TKA component. PEEK and CoCr implants were subjected to a walking gait cycle for 10 million cycles (MC), 100,000 cycles or 0 cycles (unloaded control). A method was developed to assess the fixation at the cement–implant interface, which exposed the implants to a fluorescent penetrant dye solution. The lateral condyles of the implants were then sectioned and viewed under fluorescence to investigate bonding at the cement–implant interface and cracking of the cement mantle. When tested for 100,000 cycles, debonding of the cement–implant interface occurred in both PEEK (61%) and CoCr (13%) implants. When the duration of testing was extended (10 MC), the percentage debonding was further increased for both materials to 88% and 61% for PEEK and CoCr, respectively. The unloaded PEEK specimens were 79% debonded, which suggests that, when PEEK femoral components are cemented, complete bonding may never occur. Analysis of cracks in the cement mantle showed an absence of full-thickness cracks in the unloaded control group. For the 100,000-cycle samples, on average, 1.3 and 0.7 cracks were observed for PEEK and CoCr specimens, respectively. After 10 MC, these increased to 24 for PEEK and 19 for CoCr. This was a preliminary study with a limited number of samples investigated, but shows that, after 10 MC under a walking gait, substantial debonding was visible for both PEEK and CoCr implants at the cement–implant interface and no significant difference in the number of cement cracks was found between the two materials.

ECM-based microfluidic gradient generator for tunable surface environment by interstitial flow.

We present an extracellular matrix (ECM)-based gradient generator that provides a culture surface with continuous chemical concentration gradients created by interstitial flow. The gelatin-based microchannels harboring gradient generators and in-channel micromixers were rapidly fabricated by sacrificial molding of a 3D-printed water-soluble sacrificial mold. When fluorescent dye solutions were introduced into the channel, the micromixers enhanced mixing of two solutions joined at the junction. Moreover, the concentration gradients generated in the channel diffused to the culture surface of the device through the interstitial space facilitated by the porous nature of the ECM. To check the functionality of the gradient generator for investigating cellular responses to chemical factors, we demonstrated that human umbilical vein endothelial cells cultured on the surface shrunk in response to the concentration gradient of histamine generated by interstitial flow from the microchannel. We believe that our device could be useful for the basic biological study of the cellular response to chemical stimuli and for the in vitro platform in drug testing.

Floral Traits Predict Frequency of Defecation on Flowers by Foraging Bumble Bees.

Flowers may become inoculated with pathogens that can infect bees and other critical pollinators, but the mechanisms of inoculation remain unclear. During foraging, bees may regurgitate or defecate directly onto flower parts, which could inoculate flowers with pollinator pathogens and lead to subsequent disease transmission to floral visitors. We tested if captive eastern bumble bees (Bombus impatiens) (Cresson) (Hymenoptera: Apidae) defecate on floral surfaces during foraging and if flower shape played a role in the probability of defecation and the quantity of feces deposited on floral surfaces. Captive Bombus impatiens were fed a solution of fluorescent dye and sucrose, then allowed to forage freely on flowers of a variety of shapes in a flight cage. Flowers were then examined under ultraviolet light for fluorescing fecal matter. We found that bumble bees did defecate on floral surfaces during foraging and that composite flowers with a large area of disk flowers were the most likely to have feces on them. Our results point to defecation by bumble bees during foraging as a potential mechanism for inoculation of flowers with pollinator pathogens and suggest that flower shape could play a significant role in inoculation.

Multipoint fluorescence correlation spectroscopy using spatial light modulator.

A multipoint holographic fluorescence correlation spectroscope (MP-hFCS) was successfully developed. The validity of the MP-hFCS was demonstrated using diffusion measurements of fluorescent dye solutions and of fluorescent proteins in single cells. Furthermore, the successful detection of the nuclear transport of a green fluorescent protein-tagged glucocorticoid receptor α indicates the possibility of being able to monitor directional molecular transport using the MP-hFCS. This allows multipoint analysis of the intermolecular interactions and molecular transport in living cells. Finally, the MP-hFCS can achieve multipoint diffusion measurements with high spatial and time resolution while maintaining a high photon detection sensitivity.

Calibration of an in-water multi-excitation fluorometer for the measurement of phytoplankton chlorophyll-a fluorescence quantum yield.

A multi-excitation fluorometer (MFL, JFE Advantech Co., Ltd.), originally designed to discriminate between phytoplankton species present within a population, has been redirected for use in fluorescence quantum yield (FQY) determination. While this calibration for apparent FQY requires no modification of the MFL, it is necessary to have an independent measurement of the spectral absorption coefficient of the subject fluid. Two different approaches to calibration were implemented. The primary method made use of reference fluorescent dye solutions of known quantum yield. The second method made use of acrylic fluorescent plaques and films. The two methods yielded consistent results, except in the 570 and 590 nm LED channels of the MFL. Application of the MFL in FQY determination is illustrated with an in situ Southern Ocean sample.

Investigation of controlled solvent exchange precipitation of fluorescent organic nanocrystals

The process of bottom-up organic nanocrystal precipitation is influenced by various parameters, like precursor concentration, flow rates (if using a mixing system) and presence of supporting substances, e.g. surfactants. Knowing that supporting substances are often undesired and harmful in the context of biomedical engineering and drug technology, designing an efficient, controllable and surfactant-free process is important. Organic nanocrystals can be used in delivery of poorly soluble drugs and in the optical detection of tumors when fluorescent nanocrystals are employed. Both drugs and fluorescent agents take the advantage of a small size, purity and possibilities of functionalization for targeting cancer cells. We investigated key process parameters in the precipitation of [2-[2-[4-(dimethylamino)phenyl]ethenyl]-6-methyl-4H-pyran-4-ylidene]-propanedinitrile (DCM) fluorescent nanocrystals in a hydrodynamic focusing device. We have identified the influence of fluorescent dye solution concentration and water/dye solution flow rate ratio on obtained product polydispersity, size, zeta potential, and fluorescence. We have compared the results with bulk precipitated crystals. Device-obtained nanocrystals present higher monodispersity, diameters in a range of 90–150 nm and low fluorescence compared to crude crystals. Moreover, for high flow rate ratio and high dilutions of the dye, the nanocrystals present both monodispersity and intense fluorescence, even higher than crude crystals obtained in the bulk mixing process.

Direct Observations of Graphene Dispersed in Solution by Twilight Fluorescence Microscopy.

Graphene and graphene oxide (GO) in solution were directly observed by a newly developed twilight fluorescence (TwiF) microscopy. A nanocarbon dispersion was mixed with a highly concentrated fluorescent dye solution and placed in a cell with a viewing glass at the bottom. TwiF microscopy images the nanocarbon material floating within a few hundred μm of the glass surface by utilizing two optical processes to provide a faintly illuminating backlight and visualizes GO as either a dark image by absorption and energy transfer processes or a bright image by alternation of fluorophore chemistry and autofluorescence. Individual graphene and GO sheets ranging from submicron to submillimeter widths were clearly imaged at different wavelengths, which were selectable based on the dye used. Graphene could be differentiated from GO coexisting in the same solution. Partial transparency revealed layering and network structures. Motions in tumbling flow were recognized in real time. An effect of changing the solvent and the process of adhesion on the glass surface were followed in situ.

地下水トレーサとしての蛍光染料の分析と試料溶液の保管法の検討

蛍光染料は分析や保管の過程で濃度や蛍光強度が変化する可能性があり，これが定量結果に影響を与える可能性がある。本研究では，分析や保管において蛍光濃度・強度が変化する可能性がある要因を列挙して実際にその影響を評価し，さらにその対策について検討した。分析や前処理においては，溶液のpH，溶液の温度，天然有機物の共存，ろ過が分析結果に影響を与えるが，それぞれホウ砂水溶液でのpH緩衝，サンプル温度の調整，地下水の蛍光・吸光挙動把握，適切なろ紙と使用法選定により影響を低減できる。保管においては遮光により変質する可能性を低減できるが，微生物活動による変質には留意する必要がある。

Wide-field time-gated photoluminescence microscopy for fast ultrahigh-sensitivity imaging of photoluminescent probes.

Fluorescence microscopy is a fundamental technique for the life sciences, where biocompatible and photostable photoluminescence probes in combination with fast and sensitive imaging systems are continually transforming this field. A wide-field time-gated photoluminescence microscopy system customised for ultrasensitive imaging of unique nanoruby probes with long photoluminescence lifetime is described. The detection sensitivity derived from the long photoluminescence lifetime of the nanoruby makes it possible to discriminate signals from unwanted autofluorescence background and laser backscatter by employing a time-gated image acquisition mode. This mode enabled several-fold improvement of the photoluminescence imaging contrast of discrete nanorubies dispersed on a coverslip. It enabled recovery of the photoluminescence signal emanating from discrete nanorubies when covered by a layer of an organic fluorescent dye, which were otherwise invisible without the use of spectral filtering approaches. Time-gated imaging also facilitated high sensitivity detection of nanorubies in a biological environment of cultured cells. Finally, we monitor the binding kinetics of nanorubies to a functionalised substrate, which exemplified a real-time assay in biological fluids. 3D-pseudo colour images of nanorubies immersed in a highly fluorescent dye solution. Nanoruby photoluminescence is subdued by that of the dye in continuous excitation/imaging (left), however it can be recovered by time-gated imaging (right). At the bottom is schematic diagram of nanoruby assay in a biological fluid.