Design and characterization of an optimized simultaneous color and near-infrared fluorescence rigid endoscopic imaging system

NASA Astrophysics Data System (ADS)

Venugopal, Vivek; Park, Minho; Ashitate, Yoshitomo; Neacsu, Florin; Kettenring, Frank; Frangioni, John V.; Gangadharan, Sidhu P.; Gioux, Sylvain

2013-12-01

We report the design, characterization, and validation of an optimized simultaneous color and near-infrared (NIR) fluorescence rigid endoscopic imaging system for minimally invasive surgery. This system is optimized for illumination and collection of NIR wavelengths allowing the simultaneous acquisition of both color and NIR fluorescence at frame rates higher than 6.8 fps with high sensitivity. The system employs a custom 10-mm diameter rigid endoscope optimized for NIR transmission. A dual-channel light source compatible with the constraints of an endoscope was built and includes a plasma source for white light illumination and NIR laser diodes for fluorescence excitation. A prism-based 2-CCD camera was customized for simultaneous color and NIR detection with a highly efficient filtration scheme for fluorescence imaging of both 700- and 800-nm emission dyes. The performance characterization studies indicate that the endoscope can efficiently detect fluorescence signal from both indocyanine green and methylene blue in dimethyl sulfoxide at the concentrations of 100 to 185 nM depending on the background optical properties. Finally, we performed the validation of this imaging system in vivo during a minimally invasive procedure for thoracic sentinel lymph node mapping in a porcine model.

Scalable ion-photon quantum interface based on integrated diffractive mirrors

NASA Astrophysics Data System (ADS)

Ghadimi, Moji; Blūms, Valdis; Norton, Benjamin G.; Fisher, Paul M.; Connell, Steven C.; Amini, Jason M.; Volin, Curtis; Hayden, Harley; Pai, Chien-Shing; Kielpinski, David; Lobino, Mirko; Streed, Erik W.

2017-12-01

Quantum networking links quantum processors through remote entanglement for distributed quantum information processing and secure long-range communication. Trapped ions are a leading quantum information processing platform, having demonstrated universal small-scale processors and roadmaps for large-scale implementation. Overall rates of ion-photon entanglement generation, essential for remote trapped ion entanglement, are limited by coupling efficiency into single mode fibers and scaling to many ions. Here, we show a microfabricated trap with integrated diffractive mirrors that couples 4.1(6)% of the fluorescence from a 174Yb+ ion into a single mode fiber, nearly triple the demonstrated bulk optics efficiency. The integrated optic collects 5.8(8)% of the π transition fluorescence, images the ion with sub-wavelength resolution, and couples 71(5)% of the collected light into the fiber. Our technology is suitable for entangling multiple ions in parallel and overcomes mode quality limitations of existing integrated optical interconnects.

Two-photon imaging in living brain slices.

PubMed

Mainen, Z F; Maletic-Savatic, M; Shi, S H; Hayashi, Y; Malinow, R; Svoboda, K

1999-06-01

Two-photon excitation laser scanning microscopy (TPLSM) has become the tool of choice for high-resolution fluorescence imaging in intact neural tissues. Compared with other optical techniques, TPLSM allows high-resolution imaging and efficient detection of fluorescence signal with minimal photobleaching and phototoxicity. The advantages of TPLSM are especially pronounced in highly scattering environments such as the brain slice. Here we describe our approaches to imaging various aspects of synaptic function in living brain slices. To combine several imaging modes together with patch-clamp electrophysiological recordings we found it advantageous to custom-build an upright microscope. Our design goals were primarily experimental convenience and efficient collection of fluorescence. We describe our TPLSM imaging system and its performance in detail. We present dynamic measurements of neuronal morphology of neurons expressing green fluorescent protein (GFP) and GFP fusion proteins as well as functional imaging of calcium dynamics in individual dendritic spines. Although our microscope is a custom instrument, its key advantages can be easily implemented as a modification of commercial laser scanning microscopes. Copyright 1999 Academic Press.

Contact-lens type of micromachined hydrogenated amorphous Si fluorescence detector coupled with microfluidic electrophoresis devices

NASA Astrophysics Data System (ADS)

Kamei, Toshihiro; Wada, Takehito

2006-09-01

A 5.8-μm-thick SiO2/Ta2O5 multilayer optical interference filter was monolithically integrated and micromachined on a hydrogenated amorphous Si (a-Si :H) pin photodiode to form a fluorescence detector. A microfluidic electrophoresis device was mounted on a detection platform comprising a fluorescence-collecting half-ball lens and the micromachined fluorescence detector. The central aperture of the fluorescence detector allows semiconductor laser light to pass up through the detector and to irradiate an electrophoretic separation channel. The limit of detection is as low as 7nM of the fluorescein solution, and high-speed DNA fragment sizing can be achieved with high separation efficiency. The micromachined a-Si :H fluorescence detector exhibits high sensitivity for practical fluorescent labeling dyes as well as integration flexibility on various substances, making it ideal for application to portable microfluidic bioanalysis devices.

Measuring partial fluorescence yield using filtered detectors.

PubMed

Boyko, T D; Green, R J; Moewes, A; Regier, T Z

2014-07-01

Typically, X-ray absorption near-edge structure measurements aim to probe the linear attenuation coefficient. These measurements are often carried out using partial fluorescence yield techniques that rely on detectors having photon energy discrimination improving the sensitivity and the signal-to-background ratio of the measured spectra. However, measuring the partial fluorescence yield in the soft X-ray regime with reasonable efficiency requires solid-state detectors, which have limitations due to the inherent dead-time while measuring. Alternatively, many of the available detectors that are not energy dispersive do not suffer from photon count rate limitations. A filter placed in front of one of these detectors will make the energy-dependent efficiency non-linear, thereby changing the responsivity of the detector. It is shown that using an array of filtered X-ray detectors is a viable method for measuring soft X-ray partial fluorescence yield spectra without dead-time. The feasibility of this technique is further demonstrated using α-Fe2O3 as an example and it is shown that this detector technology could vastly improve the photon collection efficiency at synchrotrons and that these detectors will allow experiments to be completed with a much lower photon flux reducing X-ray-induced damage.

Improvement of fluorescence intensity of nitrogen vacancy centers in self-formed diamond microstructures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Furuyama, S.; Yaita, J.; Kondo, M.

2015-10-19

We present umbrella-shaped diamond microstructures with metal mirrors at the bottom in order to improve the amount of collected photons from nitrogen vacancy centers. The metal mirrors at the bottom are self-aligned to the umbrella-shaped diamond microstructures which are selectively grown through holes created on a metal mask. By the finite-difference time-domain simulations, we found that the umbrella-shaped microstructures, which have an effect similar to solid immersion lens, could collect photons more efficiently than bulk or pillar-shaped microstructures. Improvement of the fluorescence intensity by factors of from 3 to 5 is shown experimentally.

Comparison of various excitation and detection schemes for dye-doped polymeric whispering gallery mode micro-lasers.

PubMed

Siegle, Tobias; Kellerer, Jonas; Bonenberger, Marielle; Krämmer, Sarah; Klusmann, Carolin; Müller, Marius; Kalt, Heinz

2018-02-05

We compare different excitation and collection configurations based on free-space optics and evanescently coupled tapered fibers for both lasing and fluorescence emission from dye-doped doped polymeric whispering gallery mode (WGM) micro-disk lasers. The focus of the comparison is on the lasing threshold and efficiency of light collection. With the aid of optical fibers, we localize the pump energy to the cavity-mode volume and reduce the necessary pump energy to achieve lasing by two orders of magnitude. When using fibers for detection, the collection efficiency is enhanced by four orders of magnitude compared to a free-space read-out perpendicular to the resonator plane. By enhancing the collection efficiency we are able to record a pronounced modulation of the dye fluorescence under continuous wave (cw) pumping conditions evoked by coupling to the WGMs. Alternatively to fibers as a collection tool, we present a read-out technique based on the detection of in-plane radiated light. We show that this method is especially beneficial in an aqueous environment as well as for size-reduced micro-lasers where radiation is strongly pronounced. Furthermore, we show that this technique allows for the assignment of transverse electric (TE) and transverse magnetic (TM) polarization to the observed fundamental modes in a water environment by performing polarization-dependent photoluminescence (PL) spectroscopy. We emphasize the importance of the polarization determination for sensing applications and verify expected differences in the bulk refractive index sensitivity for TE and TM WGMs experimentally.

A multimodal spectroscopy system for real-time disease diagnosis

NASA Astrophysics Data System (ADS)

Šćepanović, Obrad R.; Volynskaya, Zoya; Kong, Chae-Ryon; Galindo, Luis H.; Dasari, Ramachandra R.; Feld, Michael S.

2009-04-01

The combination of reflectance, fluorescence, and Raman spectroscopy—termed multimodal spectroscopy (MMS)—provides complementary and depth-sensitive information about tissue composition. As such, MMS is a promising tool for disease diagnosis, particularly in atherosclerosis and breast cancer. We have developed an integrated MMS instrument and optical fiber spectral probe for simultaneous collection of all three modalities in a clinical setting. The MMS instrument multiplexes three excitation sources, a xenon flash lamp (370-740 nm), a nitrogen laser (337 nm), and a diode laser (830 nm), through the MMS probe to excite tissue and collect the spectra. The spectra are recorded on two spectrograph/charge-coupled device modules, one optimized for visible wavelengths (reflectance and fluorescence) and the other for the near-infrared (Raman), and processed to provide diagnostic parameters. We also describe the design and calibration of a unitary MMS optical fiber probe 2 mm in outer diameter, containing a single appropriately filtered excitation fiber and a ring of 15 collection fibers, with separate groups of appropriately filtered fibers for efficiently collecting reflectance, fluorescence, and Raman spectra from the same tissue location. A probe with this excitation/collection geometry has not been used previously to collect reflectance and fluorescence spectra, and thus physical tissue models ("phantoms") are used to characterize the probe's spectroscopic response. This calibration provides probe-specific modeling parameters that enable accurate extraction of spectral parameters. This clinical MMS system has been used recently to analyze artery and breast tissue in vivo and ex vivo.

Lithographically-fabricated channel arrays for confocal x-ray fluorescence microscopy and XAFS

NASA Astrophysics Data System (ADS)

Woll, Arthur R.; Agyeman-Budu, David; Choudhury, Sanjukta; Coulthard, Ian; Finnefrock, Adam C.; Gordon, Robert; Hallin, Emil; Mass, Jennifer

2014-03-01

Confocal X-ray Fluorescence Microscopy (CXRF) employs overlapping focal regions of two x-ray optics—a condenser and collector—to directly probe a 3D volume. The minimum-achievable size of this probe volume is limited by the collector, for which polycapillaries are generally the optic of choice. Recently, we demonstrated an alternative collection optic for CXRF, consisting of an array of micron-scale collimating channels, etched in silicon, and arranged like spokes of a wheel directed towards a single source position. The optic, while successful, had a working distance of only 0.2 mm and exhibited relatively low total collection efficiency, limiting its practical application. Here, we describe a new design in which the collimating channels are formed by a staggered array of pillars whose side-walls taper away from the channel axis. This approach improves both collection efficiency and working distance, while maintaining excellent spatial resolution. We illustrate these improvements with confocal XRF data obtained at the Cornell High Energy Synchrotron Source (CHESS) and the Advanced Photon Source (APS) beamline 20-ID-B.

Molecular imaging needles: dual-modality optical coherence tomography and fluorescence imaging of labeled antibodies deep in tissue

PubMed Central

Scolaro, Loretta; Lorenser, Dirk; Madore, Wendy-Julie; Kirk, Rodney W.; Kramer, Anne S.; Yeoh, George C.; Godbout, Nicolas; Sampson, David D.; Boudoux, Caroline; McLaughlin, Robert A.

2015-01-01

Molecular imaging using optical techniques provides insight into disease at the cellular level. In this paper, we report on a novel dual-modality probe capable of performing molecular imaging by combining simultaneous three-dimensional optical coherence tomography (OCT) and two-dimensional fluorescence imaging in a hypodermic needle. The probe, referred to as a molecular imaging (MI) needle, may be inserted tens of millimeters into tissue. The MI needle utilizes double-clad fiber to carry both imaging modalities, and is interfaced to a 1310-nm OCT system and a fluorescence imaging subsystem using an asymmetrical double-clad fiber coupler customized to achieve high fluorescence collection efficiency. We present, to the best of our knowledge, the first dual-modality OCT and fluorescence needle probe with sufficient sensitivity to image fluorescently labeled antibodies. Such probes enable high-resolution molecular imaging deep within tissue. PMID:26137379

Enhanced-locality fiber-optic two-photon-fluorescence live-brain interrogation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fedotov, I. V.; Doronina-Amitonova, L. V.; Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125

2014-02-24

Two-photon excitation is shown to substantially enhance the locality of fiber-based optical interrogation of strongly scattering biotissues. In our experiments, a high-numerical-aperture, large-core-are fiber probe is used to deliver the 200-fs output of a 100-MHz mode-locked ytterbium fiber laser to samples of live mouse brain, induce two-photon fluorescence of nitrogen–vacancy centers in diamond markers in brain sample. Fiber probes with a high numerical aperture and a large core area are shown to enable locality enhancement in fiber-laser–fiber-probe two-photon brain excitation and interrogation without sacrificing the efficiency of fluorescence response collection.

An integrated centrifugo-opto-microfluidic platform for arraying, analysis, identification and manipulation of individual cells.

PubMed

Burger, R; Kurzbuch, D; Gorkin, R; Kijanka, G; Glynn, M; McDonagh, C; Ducrée, J

2015-01-21

In this work we present a centrifugal microfluidic system enabling highly efficient collective trapping and alignment of particles such as microbeads and cells, their multi-colour fluorescent detection and subsequent manipulation by optical tweezers. We demonstrate array-based capture and imaging followed by "cherry-picking" of individual particles, first for fluorescently labelled polystyrene (PS) beads and then for cells. Different cell lines are discriminated based on intracellular as well as surface-based markers.

Nanoscale volume confinement and fluorescence enhancement with double nanohole aperture

PubMed Central

Regmi, Raju; Al Balushi, Ahmed A.; Rigneault, Hervé; Gordon, Reuven; Wenger, Jérôme

2015-01-01

Diffraction ultimately limits the fluorescence collected from a single molecule, and sets an upper limit to the maximum concentration to isolate a single molecule in the detection volume. To overcome these limitations, we introduce here the use of a double nanohole structure with 25 nm gap, and report enhanced detection of single fluorescent molecules in concentrated solutions exceeding 20 micromolar. The nanometer gap concentrates the light into an apex volume down to 70 zeptoliter (10−21 L), 7000-fold below the diffraction-limited confocal volume. Using fluorescence correlation spectroscopy and time-correlated photon counting, we measure fluorescence enhancement up to 100-fold, together with local density of optical states (LDOS) enhancement of 30-fold. The distinctive features of double nanoholes combining high local field enhancement, efficient background screening and relative nanofabrication simplicity offer new strategies for real time investigation of biochemical events with single molecule resolution at high concentrations. PMID:26511149

A PDMS-based cylindrical hybrid lens for enhanced fluorescence detection in microfluidic systems.

PubMed

Lin, Bor-Shyh; Yang, Yu-Ching; Ho, Chong-Yi; Yang, Han-Yu; Wang, Hsiang-Yu

2014-02-13

Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS) to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 μg/mL and 0.05 μg/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light.

Snapshot imaging Fraunhofer line discriminator for detection of plant fluorescence

NASA Astrophysics Data System (ADS)

Gupta Roy, S.; Kudenov, M. W.

2015-05-01

Non-invasive quantification of plant health is traditionally accomplished using reflectance based metrics, such as the normalized difference vegetative index (NDVI). However, measuring plant fluorescence (both active and passive) to determine photochemistry of plants has gained importance. Due to better cost efficiency, lower power requirements, and simpler scanning synchronization, detecting passive fluorescence is preferred over active fluorescence. In this paper, we propose a high speed imaging approach for measuring passive plant fluorescence, within the hydrogen alpha Fraunhofer line at ~656 nm, using a Snapshot Imaging Fraunhofer Line Discriminator (SIFOLD). For the first time, the advantage of snapshot imaging for high throughput Fraunhofer Line Discrimination (FLD) is cultivated by our system, which is based on a multiple-image Fourier transform spectrometer and a spatial heterodyne interferometer (SHI). The SHI is a Sagnac interferometer, which is dispersion compensated using blazed diffraction gratings. We present data and techniques for calibrating the SIFOLD to any particular wavelength. This technique can be applied to quantify plant fluorescence at low cost and reduced complexity of data collection.

Real-time hyperspectral fluorescence imaging of pancreatic β-cell dynamics with the image mapping spectrometer

PubMed Central

Elliott, Amicia D.; Gao, Liang; Ustione, Alessandro; Bedard, Noah; Kester, Robert; Piston, David W.; Tkaczyk, Tomasz S.

2012-01-01

Summary The development of multi-colored fluorescent proteins, nanocrystals and organic fluorophores, along with the resulting engineered biosensors, has revolutionized the study of protein localization and dynamics in living cells. Hyperspectral imaging has proven to be a useful approach for such studies, but this technique is often limited by low signal and insufficient temporal resolution. Here, we present an implementation of a snapshot hyperspectral imaging device, the image mapping spectrometer (IMS), which acquires full spectral information simultaneously from each pixel in the field without scanning. The IMS is capable of real-time signal capture from multiple fluorophores with high collection efficiency (∼65%) and image acquisition rate (up to 7.2 fps). To demonstrate the capabilities of the IMS in cellular applications, we have combined fluorescent protein (FP)-FRET and [Ca2+]i biosensors to measure simultaneously intracellular cAMP and [Ca2+]i signaling in pancreatic β-cells. Additionally, we have compared quantitatively the IMS detection efficiency with a laser-scanning confocal microscope. PMID:22854044

Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging

NASA Astrophysics Data System (ADS)

Pian, Qi; Yao, Ruoyang; Sinsuebphon, Nattawut; Intes, Xavier

2017-07-01

Spectrally resolved fluorescence lifetime imaging and spatial multiplexing have offered information content and collection-efficiency boosts in microscopy, but efficient implementations for macroscopic applications are still lacking. An imaging platform based on time-resolved structured light and hyperspectral single-pixel detection has been developed to perform quantitative macroscopic fluorescence lifetime imaging (MFLI) over a large field of view (FOV) and multiple spectral bands simultaneously. The system makes use of three digital micromirror device (DMD)-based spatial light modulators (SLMs) to generate spatial optical bases and reconstruct N by N images over 16 spectral channels with a time-resolved capability (∼40 ps temporal resolution) using fewer than N2 optical measurements. We demonstrate the potential of this new imaging platform by quantitatively imaging near-infrared (NIR) Förster resonance energy transfer (FRET) both in vitro and in vivo. The technique is well suited for quantitative hyperspectral lifetime imaging with a high sensitivity and paves the way for many important biomedical applications.

Continuous determination of gaseous ammonia in the ambient atmosphere using fluorescence derivatization

NASA Astrophysics Data System (ADS)

Abbas, Rana; Tanner, Roger L.

A method for continuous determination of ambient ammonia levels employing o-phthalaldehyde fluorescence derivatization is described. A simplified Venturi scrubber and gas-liquid separator have been employed for reproducible measurements of ⩾ 0.1 ppb ambient ammonia with less than 2 min time resolution. The scrubbing efficiency of the ammonia gas collection system was determined to be 29 ± 1 %. During 4 d in August 1979 ambient ammonia levels at the Brookhaven National Laboratory site averaged about 1.5 ± 1.1 ppb during afternoon daylight hours.

Optical fluorescence spectroscopy to detect hepatic necrosis after normothermic ischemia: animal model

NASA Astrophysics Data System (ADS)

Romano, Renan A.; Vollet-Filho, Jose D.; Pratavieira, Sebastião.; Fernandez, Jorge L.; Kurachi, Cristina; Bagnato, Vanderlei S.; Castro-e-Silva, Orlando; Sankarankutty, Ajith K.

2015-06-01

Liver transplantation is a well-established treatment for liver failure. However, the success of the transplantation procedure depends on liver graft conditions. The tissue function evaluation during the several transplantation stages is relevant, in particular during the organ harvesting, when a decision is made concerning the viability of the graft. Optical fluorescence spectroscopy is a good option because it is a noninvasive and fast technique. A partial normothermic hepatic ischemia was performed in rat livers, with a vascular occlusion of both median and left lateral lobes, allowing circulation only for the right lateral lobe and the caudate lobe. Fluorescence spectra under excitation at 532 nm (doubled frequency Nd:YAG laser) were collected using a portable spectrometer (USB2000, Ocean Optics, USA). The fluorescence emission was collected before vascular occlusion, after ischemia, and 24 hours after reperfusion. A morphometric histology analysis was performed as the gold standard evaluation - liver samples were analyzed, and the percentage of necrotic tissue was obtained. The results showed that changes in the fluorescence emission after ischemia can be correlated with the amount of necrosis evaluated by a morphometric analysis, the Pearson correlation coefficient of the generated model was 0.90 and the root mean square error was around 20%. In this context, the laser-induced fluorescence spectroscopy technique after normothermic ischemia showed to be a fast and efficient method to differentiate ischemic injury from viable tissues.

Investigating the association between photosynthetic efficiency and generation of biophotoelectricity in autotrophic microbial fuel cells

PubMed Central

Ciniciato, Gustavo P. M. K.; Ng, Fong-Lee; Phang, Siew-Moi; Jaafar, Muhammad Musoddiq; Fisher, Adrian C.; Yunus, Kamran; Periasamy, Vengadesh

2016-01-01

Microbial fuel cells operating with autotrophic microorganisms are known as biophotovoltaic devices. It represents a great opportunity for environmentally-friendly power generation using the energy of the sunlight. The efficiency of electricity generation in this novel system is however low. This is partially reflected by the poor understanding of the bioelectrochemical mechanisms behind the electron transfer from these microorganisms to the electrode surface. In this work, we propose a combination of electrochemical and fluorescence techniques, giving emphasis to the pulse amplitude modulation fluorescence. The combination of these two techniques allow us to obtain information that can assist in understanding the electrical response obtained from the generation of electricity through the intrinsic properties related to the photosynthetic efficiency that can be obtained from the fluorescence emitted. These were achieved quantitatively by means of observed changes in four photosynthetic parameters with the bioanode generating electricity. These are the maximum quantum yield (Fv/Fm), alpha (α), light saturation coefficient (Ek) and maximum rate of electron transfer (rETRm). The relationship between the increases in the current density collected by the bioanode to the decrease of the rETRm values in the photosynthetic pathway for the two microorganisms was also discussed. PMID:27502051

Investigating the association between photosynthetic efficiency and generation of biophotoelectricity in autotrophic microbial fuel cells

NASA Astrophysics Data System (ADS)

Ciniciato, Gustavo P. M. K.; Ng, Fong-Lee; Phang, Siew-Moi; Jaafar, Muhammad Musoddiq; Fisher, Adrian C.; Yunus, Kamran; Periasamy, Vengadesh

2016-08-01

Microbial fuel cells operating with autotrophic microorganisms are known as biophotovoltaic devices. It represents a great opportunity for environmentally-friendly power generation using the energy of the sunlight. The efficiency of electricity generation in this novel system is however low. This is partially reflected by the poor understanding of the bioelectrochemical mechanisms behind the electron transfer from these microorganisms to the electrode surface. In this work, we propose a combination of electrochemical and fluorescence techniques, giving emphasis to the pulse amplitude modulation fluorescence. The combination of these two techniques allow us to obtain information that can assist in understanding the electrical response obtained from the generation of electricity through the intrinsic properties related to the photosynthetic efficiency that can be obtained from the fluorescence emitted. These were achieved quantitatively by means of observed changes in four photosynthetic parameters with the bioanode generating electricity. These are the maximum quantum yield (Fv/Fm), alpha (α), light saturation coefficient (Ek) and maximum rate of electron transfer (rETRm). The relationship between the increases in the current density collected by the bioanode to the decrease of the rETRm values in the photosynthetic pathway for the two microorganisms was also discussed.

Anomalous doping of a molecular crystal monitored with confocal fluorescence microscopy: Terrylene in a p-terphenyl crystal

NASA Astrophysics Data System (ADS)

Białkowska, Magda; Deperasińska, Irena; Makarewicz, Artur; Kozankiewicz, Bolesław

2017-09-01

Highly terrylene doped single crystals of p-terphenyl, obtained by co-sublimation of both components, showed bright spots in the confocal fluorescence images. Polarization of the fluorescence excitation spectra, blinking and bleaching, and saturation behavior allowed us to attribute them to single molecules of terrylene anomalously embedded between two neighbor layers of the host crystal, in the (a,b) plane. Such an orientation of terrylene molecules results in much more efficient absorption and collection of the fluorescence photons than in the case of previously investigated molecules embedded in the substitution sites. The above conclusion was supported by quantum chemistry calculations. We postulate that the kind of doping considered in this work should be possible in other molecular crystals where the host molecules are organized in a herringbone pattern.

Utilizing Diffuse Reflection to Increase the Efficiency of Luminescent Solar Concentrators

NASA Astrophysics Data System (ADS)

Bowser, Seth; Weible, Seth; Solomon, Joel; Schrecengost, Jonathan; Wittmershaus, Bruce

A luminescent solar concentrator (LSC) consists of a high index solid plate containing a fluorescent material that converts sunlight into fluorescence. Utilizing total internal reflection, the LSC collects and concentrates the fluorescence at the plate's edges where it is converted into electricity via photovoltaic solar cells. The lower production costs of LSCs make them an attractive alternative to photovoltaic solar cells. To optimize an LSC's efficiency, a white diffusive surface (background) is positioned behind it. The background allows sunlight transmitted in the first pass to be reflected back through the LSC providing a second chance for absorption. Our research examines how the LSC's performance is affected by changing the distance between the white background and the LSC. An automated linear motion apparatus was engineered to precisely measure this distance and the LSC's electrical current, simultaneously. LSC plates, with and without the presence of fluorescent material and in an isolated environment, showed a maximum current at a distance greater than zero. Further experimentation has proved that the optimal distance results from the background's optical properties and how the reflected light enters the LSC. This material is based upon work supported by the National Science Foundation under Grant Number NSF-ECCS-1306157.

Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses.

PubMed

Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

2016-01-01

Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination.

Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses

PubMed Central

Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

2016-01-01

Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination. PMID:26877781

Applications of Fluorescence Spectroscopy for dissolved organic matter characterization in wastewater treatment plants

NASA Astrophysics Data System (ADS)

Goffin, Angélique; Guérin, Sabrina; Rocher, Vincent; Varrault, Gilles

2016-04-01

Dissolved organic matter (DOM) influences wastewater treatment plants efficiency (WTTP): variations in its quality and quantity can induce a foaming phenomenon and a fouling event inside biofiltration processes. Moreover, in order to manage denitrification step (control and optimization of the nitrate recirculation), it is important to be able to estimate biodegradable organic matter quantity before biological treatment. But the current methods used to characterize organic matter quality, like biological oxygen demand are laborious, time consuming and sometimes not applicable to directly monitor organic matter in situ. In the context of MOCOPEE research program (www.mocopee.com), this study aims to assess the use of optical techniques, such as UV-Visible absorbance and more specifically fluorescence spectroscopy in order to monitor and to optimize process efficiency in WWTP. Fluorescence excitation-emission matrix (EEM) spectroscopy was employed to prospect the possibility of using this technology online and in real time to characterize dissolved organic matter in different effluents of the WWTP Seine Centre (240,000 m3/day) in Paris, France. 35 sewage water influent samples were collected on 10 days at different hours. Data treatment were performed by two methods: peak picking and parallel factor analysis (PARAFAC). An evolution of DOM quality (position of excitation - emission peaks) and quantity (intensity of fluorescence) was observed between the different treatment steps (influent, primary treatment, biological treatment, effluent). Correlations were found between fluorescence indicators and different water quality key parameters in the sewage influents. We developed different multivariate linear regression models in order to predict a variety of water quality parameters by fluorescence intensity at specific excitation-emission wavelengths. For example dissolved biological oxygen demand (r2=0,900; p<0,0001) and ammonium concentration (r2=0,898; p<0,0001) present good correlation with specific fluorescence peaks and indicators. These indicators derived from 3D spectrofluorescence could be used in order to characterize DOM online and thus to optimize process efficiency in WWTP.

A matter of collection and detection for intraoperative and noninvasive near-infrared fluorescence molecular imaging: To see or not to see?

PubMed Central

Zhu, Banghe; Rasmussen, John C.; Sevick-Muraca, Eva M.

2014-01-01

Purpose: Although fluorescence molecular imaging is rapidly evolving as a new combinational drug/device technology platform for molecularly guided surgery and noninvasive imaging, there remains no performance standards for efficient translation of “first-in-humans” fluorescent imaging agents using these devices. Methods: The authors employed a stable, solid phantom designed to exaggerate the confounding effects of tissue light scattering and to mimic low concentrations (nM–pM) of near-infrared fluorescent dyes expected clinically for molecular imaging in order to evaluate and compare the commonly used charge coupled device (CCD) camera systems employed in preclinical studies and in human investigational studies. Results: The results show that intensified CCD systems offer greater contrast with larger signal-to-noise ratios in comparison to their unintensified CCD systems operated at clinically reasonable, subsecond acquisition times. Conclusions: Camera imaging performance could impact the success of future “first-in-humans” near-infrared fluorescence imaging agent studies. PMID:24506637

A matter of collection and detection for intraoperative and noninvasive near-infrared fluorescence molecular imaging: To see or not to see?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhu, Banghe; Rasmussen, John C.; Sevick-Muraca, Eva M., E-mail: Eva.Sevick@uth.tmc.edu

2014-02-15

Purpose: Although fluorescence molecular imaging is rapidly evolving as a new combinational drug/device technology platform for molecularly guided surgery and noninvasive imaging, there remains no performance standards for efficient translation of “first-in-humans” fluorescent imaging agents using these devices. Methods: The authors employed a stable, solid phantom designed to exaggerate the confounding effects of tissue light scattering and to mimic low concentrations (nM–pM) of near-infrared fluorescent dyes expected clinically for molecular imaging in order to evaluate and compare the commonly used charge coupled device (CCD) camera systems employed in preclinical studies and in human investigational studies. Results: The results show thatmore » intensified CCD systems offer greater contrast with larger signal-to-noise ratios in comparison to their unintensified CCD systems operated at clinically reasonable, subsecond acquisition times. Conclusions: Camera imaging performance could impact the success of future “first-in-humans” near-infrared fluorescence imaging agent studies.« less

Optical Sensing of Ecosystem Carbon Fluxes Combining Spectral Reflectance Indices with Solar Induced Fluorescence

NASA Astrophysics Data System (ADS)

Huemmrich, K. F.; Middleton, E.; Corp, L. A.; Campbell, P. K.; Kustas, W. P.

2014-12-01

Optical sampling of spectral reflectance and solar induced fluorescence provide information on the physiological status of vegetation that can be used to infer stress responses and estimates of production. Multiple repeated observations are required to observe the effects of changing environmental conditions on vegetation. This study examines the use of optical signals to determine inputs to a light use efficiency (LUE) model describing productivity of a cornfield where repeated observations of carbon flux, spectral reflectance and fluorescence were collected. Data were collected at the Optimizing Production Inputs for Economic and Environmental Enhancement (OPE3) fields (39.03°N, 76.85°W) at USDA Beltsville Agricultural Research Center. Agricultural Research Service researchers measured CO2 fluxes using eddy covariance methods throughout the growing season. Optical measurements were made from the nearby tower supporting the NASA FUSION sensors. The sensor system consists of two dual channel, upward and downward looking, spectrometers used to simultaneously collect high spectral resolution measurements of reflected and fluoresced light from vegetation canopies. Estimates of chlorophyll fluorescence, combined with measures of vegetation pigment content and the Photosynthetic Reflectance Index (PRI) derived from the spectral reflectance are compared with CO2 fluxes over diurnal periods for multiple days. PRI detects changes in Xanthophyll cycle pigments using reflectance at 531 nm compared to a reference band at 570 nm. The relationships among the different optical measurements indicate that they are providing different types of information on the vegetation and that combinations of these measurements provide improved retrievals of CO2 fluxes than any index alone.

Optical Sensing of Ecosystem Carbon Fluxes Combining Spectral Reflectance Indices with Solar Induced Fluorescence

NASA Astrophysics Data System (ADS)

Huemmrich, K. F.; Corp, L.; Campbell, P. K.; Cook, B. D.; Middleton, E.; Cheng, Y.; Zhang, Q.; Russ, A.; Kustas, W. P.

2013-12-01

Optical sampling of spectral reflectance and solar induced fluorescence provide information on the physiological status of vegetation that can be used to infer stress responses and estimates of production. Multiple repeated observations can observe the effects of changing environmental conditions on vegetation. This study examines the use of optical signals to determine inputs to a light use efficiency (LUE) model describing productivity of a cornfield where repeated observations of carbon flux, spectral reflectance and fluorescence were collected. Data were collected at the Optimizing Production Inputs for Economic and Environmental Enhancement (OPE3) fields (39.03°N, 76.85°W) at USDA Beltsville Agricultural Research Center. Agricultural Research Service researchers measured CO2 fluxes using eddy covariance methods throughout the growing season. Optical measurements were made from the nearby tower supporting the NASA FUSION sensors. This sensor system consists of two dual channel, upward and downward looking, spectrometers used to simultaneously collect high spectral resolution measurements of reflected and fluoresced light from vegetation canopies. Estimates of chlorophyll fluorescence, combined with measures of vegetation pigment content and the Photosynthetic Reflectance Index (PRI) derived from the spectral reflectance are compared with CO2 fluxes over diurnal periods for multiple days. PRI detects changes in Xanthophyll cycle pigments using reflectance at 531 nm compared to a reference band at 570 nm. The relationships among the different optical measurements indicate that they are providing different types of information on the vegetation and that combinations of these measurements provide improved retrievals of CO2 fluxes than any index alone.

Donor-Acceptor-Collector Ternary Crystalline Films for Efficient Solid-State Photon Upconversion.

PubMed

Ogawa, Taku; Hosoyamada, Masanori; Yurash, Brett; Nguyen, Thuc-Quyen; Yanai, Nobuhiro; Kimizuka, Nobuo

2018-06-25

It is pivotal to achieve efficient triplet-triplet annihilation based photon upconversion (TTA-UC) in the solid-state for enhancing potentials of renewable energy production devices. However, the UC efficiency of solid materials is largely limited by low fluorescence quantum yields that originate from the aggregation of TTA-UC chromophores, and also by severe back energy transfer from the acceptor singlet state to the singlet state of the triplet donor in the condensed state. In this work, to overcome these issues, we introduce a highly fluorescent singlet energy collector as the third component of donor-doped acceptor crystalline films, in which dual energy migration, i.e., triplet energy migration for TTA-UC and succeeding singlet energy migration for transferring energy to a collector, takes place. To demonstrate this scheme, a highly fluorescent singlet energy collector was added as the third component of donor-doped acceptor crystalline films. An anthracene-based acceptor containing alkyl chains and a carboxylic moiety is mixed with the triplet donor Pt(II) octaethylporphyrin (PtOEP) and the energy collector 2,5,8,11-tetra- tert-butylperylene (TTBP) in solution, and spin-coating of the mixed solution gives acceptor films of nanofibrous crystals homogeneously doped with PtOEP and TTBP. Interestingly, delocalized singlet excitons in acceptor crystals are found to diffuse effectively over the distance of ~37 nm. Thanks to this high diffusivity, only 0.5 mol% of doped TTBP can harvest most of the singlet excitons, which successfully doubles the solid-state fluorescent quantum yield of acceptor/TTBP blend films to 76%. Furthermore, since the donor PtOEP and the collector TTBP are separately isolated in the nanofibrous acceptor crystals, the singlet back energy transfer from the collector to the donor is effectively avoided. Such efficient singlet energy collection and inhibited back energy transfer processes result in a large increase of UC efficiency up to 9.0%, offering rational design principles towards ultimately efficient solid-state upconverters.

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence.

PubMed

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2012-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes.

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence

PubMed Central

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2013-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes. PMID:24179476

Efficient triplet harvesting by fluorescent molecules through exciplexes for high efficiency organic light-emitting diodes

NASA Astrophysics Data System (ADS)

Park, Young-Seo; Kim, Kwon-Hyeon; Kim, Jang-Joo

2013-04-01

Efficient triplet harvesting from exciplexes by reverse intersystem crossing (RISC) is reported using a fluorescent molecular system composed of the 4,4',4″-tris(N-carbazolyl)-triphenylamine and bis-4,6-(3,5-di-3-pyridylphenyl)-2-methylpyrimidine. The exciplex forming material system shows the efficient delayed fluorescence emission. As a result, almost 100% PL efficiency at 35 K and 10% external quantum efficiency at 195 K are achieved from the exciplex. The delayed fluorescence of the exciplex clearly demonstrates that a significant proportion of the triplet exciplexes is harvested through the RISC.

Rapid and qualitative fluorescence-based method for the assessment of PHA production in marine bacteria during batch culture.

PubMed

Elain, Anne; Le Fellic, Magali; Corre, Yves-Marie; Le Grand, Adélaïde; Le Tilly, Véronique; Audic, Jean-Luc; Bruzaud, Stéphane

2015-10-01

The expansion of polyhydroxyalkanoates (PHAs) into the biodegradable polymers market is mainly prevented by their production process which is still complicated with a low efficiency, resulting in relatively expensive products. In this study, we developed a method that used the lipophilic fluorescent probe Nile Red (1 mg l(-1) solution in DMSO) directly into the culture broth to stain the PHA inclusions inside bacterial cells followed by detection of the emitted fluorescence by both microscopic and spectrometric techniques. Epifluorescence microscopy provides a rapid tool to distinguish producing from non-producing bacterial species and the relative fluorescence intensity (FI) determined at the maximum of emission spectra in the wavelength region of 560-710 nm (λ(ex): 543 nm), allows a fast assessment of the cultural conditions that may enhance PHA production yield. During two-step cultivation in 500-ml flasks with glucose as the sole carbon source, the method aimed to select bacterial strains efficient for PHA synthesis among a marine collection. Subsequently, the NR assay was used to determine the C0/N0 ratio of the producing media that may improve the polymer yield as well as to follow the time course of fermentation. Characterization by GC-MS and DSC confirmed the production of the P(3-HB) homopolymer.

Utilizing a Tower Based System for Optical Sensing of Ecosystem Carbon Fluxes

NASA Astrophysics Data System (ADS)

Huemmrich, K. F.; Corp, L. A.; Middleton, E.; Campbell, P. K. E.; Landis, D.; Kustas, W. P.

2015-12-01

Optical sampling of spectral reflectance and solar induced fluorescence provide information on the physiological status of vegetation that can be used to infer stress responses and estimates of production. Multiple repeated observations are required to observe the effects of changing environmental conditions on vegetation. This study examines the use of optical signals to determine inputs to a light use efficiency (LUE) model describing productivity of a cornfield where repeated observations of carbon flux, spectral reflectance and fluorescence were collected. Data were collected at the Optimizing Production Inputs for Economic and Environmental Enhancement (OPE3) fields (39.03°N, 76.85°W) at USDA Beltsville Agricultural Research Center. Agricultural Research Service researchers measured CO2 fluxes using eddy covariance methods throughout the growing season. Optical measurements were made from the nearby tower supporting the NASA FUSION sensors. The sensor system consists of two dual channel, upward and downward looking, spectrometers used to simultaneously collect high spectral resolution measurements of reflected and fluoresced light from vegetation canopies at multiple view angles. Estimates of chlorophyll fluorescence, combined with measures of vegetation pigment content and the Photosynthetic Reflectance Index (PRI) derived from the spectral reflectance are compared with CO2 fluxes over diurnal periods for multiple days. The relationships among the different optical measurements indicate that they are providing different types of information on the vegetation and that combinations of these measurements provide improved retrievals of CO2 fluxes than any index alone

Optical fiber-based setup for in vivo measurement of the delayed fluorescence lifetime of oxygen sensors

NASA Astrophysics Data System (ADS)

Piffaretti, Filippo M.; Santhakumar, Kanappan; Forte, Eddy; van den Bergh, Hubert E.; Wagnières, Georges A.

2011-03-01

A new optical-fiber-based spectrofluorometer for in vivo or in vitro detection of delayed fluorescence is presented and characterized. This compact setup is designed so that it can be readily adapted for future clinical use. Optical excitation is done with a nitrogen laser-pumped, tunable dye laser, emitting in the UV-vis part of the spectrum. Excitation and luminescence signals are carried to and from the biological tissues under investigation, located out of the setup enclosure, by a single optical fiber. These measurements, as well as measurements performed without a fiber on in vitro samples in a thermostable quartz cell, in a controlled-atmosphere enclosure, are possible due to the efficient collection of the laser-induced luminescence light which is collected and focused on the detector with a high aperture parabolic mirror. The detection is based on a gated photomultiplier which allows for time-resolved measurements of the delayed fluorescence intensity. Thus, relevant luminescence lifetimes, typically in the sub-microsecond-to-millisecond range, can be measured with near total rejection of the sample's prompt fluorescence. The instrument spectral and temporal resolution, as well as its sensitivity, is characterized and measurement examples are presented. The primary application foreseen for this setup is the monitoring and adjustment of the light dose delivered during photodynamic therapy.

Compact instrument for fluorescence image-guided surgery

NASA Astrophysics Data System (ADS)

Wang, Xinghua; Bhaumik, Srabani; Li, Qing; Staudinger, V. Paul; Yazdanfar, Siavash

2010-03-01

Fluorescence image-guided surgery (FIGS) is an emerging technique in oncology, neurology, and cardiology. To adapt intraoperative imaging for various surgical applications, increasingly flexible and compact FIGS instruments are necessary. We present a compact, portable FIGS system and demonstrate its use in cardiovascular mapping in a preclinical model of myocardial ischemia. Our system uses fiber optic delivery of laser diode excitation, custom optics with high collection efficiency, and compact consumer-grade cameras as a low-cost and compact alternative to open surgical FIGS systems. Dramatic size and weight reduction increases flexibility and access, and allows for handheld use or unobtrusive positioning over the surgical field.

Highly efficient phosphorescent, TADF, and fluorescent OLEDs (Conference Presentation)

NASA Astrophysics Data System (ADS)

Kim, Jang-Joo; Kim, Kwon-Hyeon; Moon, Chang-Ki; Shin, Hyun

2016-09-01

High efficiency OLEDs based on phosphorescent, thermally activated delayed fluorescent (TADF) and fluorescent emitters will be presented. We will show that EQEs over 60% is achievable if OLEDs are fabricated using organic semiconductors with the refractive indices of 1.5 and fully horizontal emitting dipoles without any extra light extracting structure. We will also show that reverse intersystem crossing RISC rate plays an important role to reduce the efficiency roll-off in efficient TADF and fluorescent OLEDs and a couple to methods will be presented to increase the RISC rate in the devices.

Epi-Fluorescence Microscopy

PubMed Central

Webb, Donna J.; Brown, Claire M.

2012-01-01

Epi-fluorescence microscopy is available in most life sciences research laboratories, and when optimized can be a central laboratory tool. In this chapter, the epi-fluorescence light path is introduced and the various components are discussed in detail. Recommendations are made for incident lamp light sources, excitation and emission filters, dichroic mirrors, objective lenses, and charge-coupled device (CCD) cameras in order to obtain the most sensitive epi-fluorescence microscope. The even illumination of metal-halide lamps combined with new “hard” coated filters and mirrors, a high resolution monochrome CCD camera, and a high NA objective lens are all recommended for high resolution and high sensitivity fluorescence imaging. Recommendations are also made for multicolor imaging with the use of monochrome cameras, motorized filter turrets, individual filter cubes, and corresponding dyes that are the best choice for sensitive, high resolution multicolor imaging. Images should be collected using Nyquist sampling and should be corrected for background intensity contributions and nonuniform illumination across the field of view. Photostable fluorescent probes and proteins that absorb a lot of light (i.e., high extinction co-efficients) and generate a lot of fluorescence signal (i.e., high quantum yields) are optimal. A neuronal immune-fluorescence labeling protocol is also presented. Finally, in order to maximize the utility of sensitive wide-field microscopes and generate the highest resolution images with high signal-to-noise, advice for combining wide-field epi-fluorescence imaging with restorative image deconvolution is presented. PMID:23026996

Etching-dependent fluorescence quenching of Ag-dielectric-Au three-layered nanoshells: The effect of inner Ag nanosphere

NASA Astrophysics Data System (ADS)

Zhu, Jian; Xu, Zai-jie; Weng, Guo-jun; Zhao, Jing; Li, Jian-jun; Zhao, Jun-wu

2018-07-01

In this report, Ag-dielectric-Au three-layered nanoshells with controlled inner core size were synthesized. The fluorescence emission of the rhodamine 6G (R6G) could be quenched by the three-layered nanoshells distinctly. What's more, the fluorescence quenching efficiency could be further improved by tuning the etching of inner Ag nanosphere. The maximum fluorescence quenching efficiency is obtained when the separate layer just appears between the inner Ag core and the outer Au shell. Whereas the fluorescence quenching efficiency is weakened when no gaps take place around the inner Ag core or the separate layer is too thick and greater than 13 nm. The fluorescence quenching properties of the Ag-dielectric-Au three-layered nanoshells with different initial sizes of the Ag nanoparticles are also studied. The maximum fluorescence quenching efficiency is obtained when the three-layered nanoshells are synthesized based on the Ag nanoparticles with 60 nm, which is better than others two sizes (42 and 79 nm). Thus we believe that the size of initial Ag nanospheres also greatly affects the optimized fluorescence quenching efficiency. These results about fluorescence quenching properties of Ag-dielectric-Au three-layered nanoshells present a potential for design and fabrication of fluorescence nanosensors based on tuning the geometry of the inner core and the separate layer.

Detection of reactive oxygen species in mainstream cigarette smoke by a fluorescent probe

NASA Astrophysics Data System (ADS)

Liu, Li; Xu, Shi-jie; Li, Song-zhan

2009-07-01

A mass of reactive oxygen species(ROS) are produced in the process of smoking. Superfluous ROS can induce the oxidative stress in organism, which will cause irreversible damage to cells. Fluorescent probe is taken as a marker of oxidative stress in biology and has been applied to ROS detection in the field of biology and chemistry for high sensitivity, high simplicity of data collection and high resolution. As one type of fluorescent probe, dihydrorhodamine 6G (dR6G) will be oxidized to the fluorescent rhodamine 6G, which could be used to detect ROS in mainstream cigarette smoke. We investigated the action mechanism of ROS on dR6G, built up the standard curve of R6G fluorescence intensity with its content, achieved the variation pattern of R6G fluorescence intensity with ROS content in mainstream cigarette smoke and detected the contents of ROS from the 4 types of cigarettes purchased in market. The result shows that the amount of ROS has close relationship with the types of tobacco and cigarette production technology. Compared with other detecting methods such as electronic spin resonance(ESR), chromatography and mass spectrometry, this detection method by the fluorescent probe has higher efficiency and sensitivity and will have wide applications in the ROS detection field.

Evaluation of a novel personal nanoparticle sampler.

PubMed

Zhou, Yue; Irshad, Hammad; Tsai, Chuen-Jinn; Hung, Shao-Ming; Cheng, Yung-Sung

2014-02-01

This work investigated the performance in terms of collection efficiency and aspiration efficiency of a personal sampler capable of collecting ultrafine particles (nanoparticles) in the occupational environment. This sampler consists of a cyclone for respirable particle classification, micro-orifice impactor stages with an acceleration nozzle to achieve nanoparticle classification and a backup filter to collect nanoparticles. Collection efficiencies of the cyclone and impactor stages were determined using monodisperse polystyrene latex and silver particles, respectively. Calibration of the cyclone and impactor stages showed 50% cut-off diameters of 3.95 μm and 94.7 nm meeting the design requirements. Aspiration efficiencies of the sampler were tested in a wind tunnel with wind speeds of 0.5, 1.0, and 1.5 m s(-1). The test samplers were mounted on a full size mannequin with three orientations toward the wind direction (0°, 90°, and 180°). Monodisperse oleic acid aerosols tagged with sodium fluorescein in the size range of 2 to 10 μm were used in the test. For particles smaller than 2 μm, the fluorescent polystyrene latex particles were generated by using nebulizers. For comparison of the aspiration efficiency, a NIOSH two-stage personal bioaerosol sampler was also tested. Results showed that the orientation-averaged aspiration efficiency for both samplers was close to the inhalable fraction curve. However, the direction of wind strongly affected the aspiration efficiency. The results also showed that the aspiration efficiency was not affected by the ratio of free-stream velocity to the velocity through the sampler orifice. Our evaluation showed that the current design of the personal sampler met the designed criteria for collecting nanoparticles ≤100 nm in occupational environments.

Simplified, rapid, and inexpensive estimation of water primary productivity based on chlorophyll fluorescence parameter Fo.

PubMed

Chen, Hui; Zhou, Wei; Chen, Weixian; Xie, Wei; Jiang, Liping; Liang, Qinlang; Huang, Mingjun; Wu, Zongwen; Wang, Qiang

2017-04-01

Primary productivity in water environment relies on the photosynthetic production of microalgae. Chlorophyll fluorescence is widely used to detect the growth status and photosynthetic efficiency of microalgae. In this study, a method was established to determine the Chl a content, cell density of microalgae, and water primary productivity by measuring chlorophyll fluorescence parameter Fo. A significant linear relationship between chlorophyll fluorescence parameter Fo and Chl a content of microalgae, as well as between Fo and cell density, was observed under pure-culture conditions. Furthermore, water samples collected from natural aquaculture ponds were used to validate the correlation between Fo and water primary productivity, which is closely related to Chl a content in water. Thus, for a given pure culture of microalgae or phytoplankton (mainly microalgae) in aquaculture ponds or other natural ponds for which the relationship between the Fo value and Chl a content or cell density could be established, Chl a content or cell density could be determined by measuring the Fo value, thereby making it possible to calculate the water primary productivity. It is believed that this method can provide a convenient way of efficiently estimating the primary productivity in natural aquaculture ponds and bringing economic value in limnetic ecology assessment, as well as in algal bloom monitoring. Copyright © 2017 Elsevier GmbH. All rights reserved.

Rise-Time of FRET-Acceptor Fluorescence Tracks Protein Folding

PubMed Central

Lindhoud, Simon; Westphal, Adrie H.; van Mierlo, Carlo P. M.; Visser, Antonie J. W. G.; Borst, Jan Willem

2014-01-01

Uniform labeling of proteins with fluorescent donor and acceptor dyes with an equimolar ratio is paramount for accurate determination of Förster resonance energy transfer (FRET) efficiencies. In practice, however, the labeled protein population contains donor-labeled molecules that have no corresponding acceptor. These FRET-inactive donors contaminate the donor fluorescence signal, which leads to underestimation of FRET efficiencies in conventional fluorescence intensity and lifetime-based FRET experiments. Such contamination is avoided if FRET efficiencies are extracted from the rise time of acceptor fluorescence upon donor excitation. The reciprocal value of the rise time of acceptor fluorescence is equal to the decay rate of the FRET-active donor fluorescence. Here, we have determined rise times of sensitized acceptor fluorescence to study the folding of double-labeled apoflavodoxin molecules and show that this approach tracks the characteristics of apoflavodoxinʼs complex folding pathway. PMID:25535076

Real-time PCR detection of Plasmodium directly from whole blood and filter paper samples

PubMed Central

2011-01-01

Background Real-time PCR is a sensitive and specific method for the analysis of Plasmodium DNA. However, prior purification of genomic DNA from blood is necessary since PCR inhibitors and quenching of fluorophores from blood prevent efficient amplification and detection of PCR products. Methods Reagents designed to specifically overcome PCR inhibition and quenching of fluorescence were evaluated for real-time PCR amplification of Plasmodium DNA directly from blood. Whole blood from clinical samples and dried blood spots collected in the field in Colombia were tested. Results Amplification and fluorescence detection by real-time PCR were optimal with 40× SYBR® Green dye and 5% blood volume in the PCR reaction. Plasmodium DNA was detected directly from both whole blood and dried blood spots from clinical samples. The sensitivity and specificity ranged from 93-100% compared with PCR performed on purified Plasmodium DNA. Conclusions The methodology described facilitates high-throughput testing of blood samples collected in the field by fluorescence-based real-time PCR. This method can be applied to a broad range of clinical studies with the advantages of immediate sample testing, lower experimental costs and time-savings. PMID:21851640

Photonic Crystal Enhanced Fluorescence for Early Breast Cancer Biomarker Detection

PubMed Central

Cunningham, Brian T.; Zangar, Richard C.

2013-01-01

Photonic crystal surfaces offer a compelling platform for improving the sensitivity of surface-based fluorescent assays used in disease diagnostics. Through the complementary processes of photonic crystal enhanced excitation and enhanced extraction, a periodic dielectric-based nanostructured surface can simultaneously increase the electric field intensity experienced by surface-bound fluorophores and increase the collection efficiency of emitted fluorescent photons. Through the ability to inexpensively fabricate photonic crystal surfaces over substantial surface areas, they are amenable to single-use applications in biological sensing, such as disease biomarker detection in serum. In this review, we will describe the motivation for implementing high-sensitivity, multiplexed biomarker detection in the context of breast cancer diagnosis. We will summarize recent efforts to improve the detection limits of such assays though the use of photonic crystal surfaces. Reduction of detection limits is driven by low autofluorescent substrates for photonic crystal fabrication, and detection instruments that take advantage of their unique features. PMID:22736539

Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo.

PubMed

Engelbrecht, Christoph J; Johnston, Richard S; Seibel, Eric J; Helmchen, Fritjof

2008-04-14

We present a small, lightweight two-photon fiberscope and demonstrate its suitability for functional imaging in the intact brain. Our device consists of a hollow-core photonic crystal fiber for efficient delivery of near-IR femtosecond laser pulses, a spiral fiber-scanner for resonant beam steering, and a gradient-index lens system for fluorescence excitation, dichroic beam splitting, and signal collection. Fluorescence light is remotely detected using a standard photomultiplier tube. All optical components have 1 mm dimensions and the microscope's headpiece weighs only 0.6 grams. The instrument achieves micrometer resolution at frame rates of typically 25 Hz with a field-of-view of up to 200 microns. We demonstrate functional imaging of calcium signals in Purkinje cell dendrites in the cerebellum of anesthetized rats. The microscope will be easily portable by a rat or mouse and thus should enable functional imaging in freely behaving animals.

Ligand Assisted Stabilization of Fluorescence Nanoparticles; an Insight on the Fluorescence Characteristics, Dispersion Stability and DNA Loading Efficiency of Nanoparticles.

PubMed

Rhouati, Amina; Hayat, Akhtar; Mishra, Rupesh K; Bueno, Diana; Shahid, Shakir Ahmad; Muñoz, Roberto; Marty, Jean Louis

2016-07-01

This work reports on the ligand assisted stabilization of Fluospheres® carboxylate modified nanoparticles (FCMNPs), and subsequently investigation on the DNA loading capacity and fluorescence response of the modified particles. The designed fluorescence bioconjugate was characterized with enhanced fluorescence characteristics, good stability and large surface area with high DNA loading efficiency. For comparison purpose, bovine serum albumin (BSA) and polyethylene glycol (PEG) with three different length strands were used as cross linkers to modify the particles, and their DNA loading capacity and fluorescence characteristics were investigated. By comparing the performance of the particles, we found that the most improved fluorescence characteristics, enhanced DNA loading and high dispersion stability were obtained, when employing PEG of long spacer arm length. The designed fluorescence bioconjugate was observed to maintain all its characteristics under varying pH over an extended period of time. These types of bioconjugates are in great demand for fluorescence imaging and in vivo fluorescence biomedical application, especially when most of the as synthesized fluorescence particles cannot withstand to varying in vivo physiological conditions with decreases in fluorescence response and DNA loading efficiency.

Rapid protein concentration, efficient fluorescence labeling and purification on a micro/nanofluidics chip.

PubMed

Wang, Chen; Ouyang, Jun; Ye, De-Kai; Xu, Jing-Juan; Chen, Hong-Yuan; Xia, Xing-Hua

2012-08-07

Fluorescence analysis has proved to be a powerful detection technique for achieving single molecule analysis. However, it usually requires the labeling of targets with bright fluorescent tags since most chemicals and biomolecules lack fluorescence. Conventional fluorescence labeling methods require a considerable quantity of biomolecule samples, long reaction times and extensive chromatographic purification procedures. Herein, a micro/nanofluidics device integrating a nanochannel in a microfluidics chip has been designed and fabricated, which achieves rapid protein concentration, fluorescence labeling, and efficient purification of product in a miniaturized and continuous manner. As a demonstration, labeling of the proteins bovine serum albumin (BSA) and IgG with fluorescein isothiocyanate (FITC) is presented. Compared to conventional methods, the present micro/nanofluidics device performs about 10(4)-10(6) times faster BSA labeling with 1.6 times higher yields due to the efficient nanoconfinement effect, improved mass, and heat transfer in the chip device. The results demonstrate that the present micro/nanofluidics device promises rapid and facile fluorescence labeling of small amount of reagents such as proteins, nucleic acids and other biomolecules with high efficiency.

A high-throughput direct fluorescence resonance energy transfer-based assay for analyzing apoptotic proteases using flow cytometry and fluorescence lifetime measurements.

PubMed

Suzuki, Miho; Sakata, Ichiro; Sakai, Takafumi; Tomioka, Hiroaki; Nishigaki, Koichi; Tramier, Marc; Coppey-Moisan, Maïté

2015-12-15

Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

Method Of Signal Amplification In Multi-Chromophore Luminescence Sensors

DOEpatents

Levitsky, Igor A.; Krivoshlykov, Sergei G.

2004-02-03

A fluorescence-based method for highly sensitive and selective detection of analyte molecules is proposed. The method employs the energy transfer between two or more fluorescent chromophores in a carefully selected polymer matrix. In one preferred embodiment, signal amplification has been achieved in the fluorescent sensing of dimethyl methylphosphonate (DMMP) using two dyes, 3-aminofluoranthene (AM) and Nile Red (NR), in a hydrogen bond acidic polymer matrix. The selected polymer matrix quenches the fluorescence of both dyes and shifts dye emission and absorption spectra relative to more inert matrices. Upon DMMP sorption, the AM fluorescence shifts to the red at the same time the NR absorption shifts to the blue, resulting in better band overlap and increased energy transfer between chromophores. In another preferred embodiment, the sensitive material is incorporated into an optical fiber system enabling efficient excitation of the dye and collecting the fluorescent signal form the sensitive material on the remote end of the system. The proposed method can be applied to multichromophore luminescence sensor systems incorporating N-chromophores leading to N-fold signal amplification and improved selectivity. The method can be used in all applications where highly sensitive detection of basic gases, such as dimethyl methylphosphonate (DMMP), Sarin, Soman and other chemical warfare agents having basic properties, is required, including environmental monitoring, chemical industry and medicine.

Spatial fluorescence cross-correlation spectroscopy between core and ring pinholes

NASA Astrophysics Data System (ADS)

Blancquaert, Yoann; Delon, Antoine; Derouard, Jacques; Jaffiol, Rodolphe

2006-04-01

Fluorescence Correlation Spectroscopy (FCS) is an attractive method to measure molecular concentration, mobility parameters and chemical kinetics. However its ability to descriminate different diffusing species needs to be improved. Recently, we have proposed a simplified spatial Fluorescence cross Correlation Spectroscopy (sFCCS) method, allowing, with only one focused laser beam to obtain two confocal volumes spatially shifted. Now, we present a new sFCCS optical geometry where the two pinholes, a ring and core, are encapsulated one in the other. In this approach all physical and chemical processes that occur in a single volume, like singlet-triplet dynamics and photobleaching, can be eliminated; moreover, this new optical geometry optimises the collection of fluorescence. The first cross Correlation curves for Rhodamine 6G (Rh6G) in Ethanol are presented, in addition to the effect of the size of fluorescent particules (nano-beads, diameters : 20, 100 and 200 nm). The relative simplicity of the method leads us to propose sFCCS as an appropriate method for the determination of diffusion parameters of fluorophores in solution or cells. Nevertheless, progresses in the ingeniering of the optical Molecular Detection Efficiency volumes are highly desirable, in order to improve the descrimination between the cross correlated volumes.

Experimental feasibility of the airborne measurement of absolute oil fluorescence spectral conversion efficiency

NASA Technical Reports Server (NTRS)

Hoge, F. E.; Swift, R. N.

1983-01-01

Airborne lidar oil spill experiments carried out to determine the practicability of the AOFSCE (absolute oil fluorescence spectral conversion efficiency) computational model are described. The results reveal that the model is suitable over a considerable range of oil film thicknesses provided the fluorescence efficiency of the oil does not approach the minimum detection sensitivity limitations of the lidar system. Separate airborne lidar experiments to demonstrate measurement of the water column Raman conversion efficiency are also conducted to ascertain the ultimate feasibility of converting such relative oil fluorescence to absolute values. Whereas the AOFSCE model is seen as highly promising, further airborne water column Raman conversion efficiency experiments with improved temporal or depth-resolved waveform calibration and software deconvolution techniques are thought necessary for a final determination of suitability.

Theoretical investigation of confocal microscopy using an elliptically polarized cylindrical vector laser beam: Visualization of quantum emitters near interfaces

NASA Astrophysics Data System (ADS)

Boichenko, Stepan

2018-04-01

We theoretically study laser-scanning confocal fluorescence microscopy using elliptically polarized cylindrical vector excitation light as a tool for visualization of arbitrarily oriented single quantum dipole emitters located (1) near planar surfaces enhancing fluorescence, (2) in a thin supported polymer film, (3) in a freestanding polymer film, and (4) in a dielectric planar microcavity. It is shown analytically that by using a tightly focused azimuthally polarized beam, it is possible to exclude completely the orientational dependence of the image intensity maximum of a quantum emitter that absorbs light as a pair of incoherent independent linear dipoles. For linear dipole quantum emitters, the orientational independence degree higher than 0.9 can normally be achieved (this quantity equal to 1 corresponds to completely excluded orientational dependence) if the collection efficiency of the microscope objective and the emitter's total quantum yield are not strongly orientationally dependent. Thus, the visualization of arbitrarily oriented single quantum emitters by means of the studied technique can be performed quite efficiently.

Far-Red Fluorescent Lipid-Polymer Probes for an Efficient Labeling of Enveloped Viruses.

PubMed

Lacour, William; Adjili, Salim; Blaising, Julie; Favier, Arnaud; Monier, Karine; Mezhoud, Sarra; Ladavière, Catherine; Place, Christophe; Pécheur, Eve-Isabelle; Charreyre, Marie-Thérèse

2016-08-01

Far-red emitting fluorescent lipid probes are desirable to label enveloped viruses, for their efficient tracking by optical microscopy inside autofluorescent cells. Most used probes are rapidly released from membranes, leading to fluorescence signal decay and loss of contrast. Here, water-soluble lipid-polymer probes are synthesized harboring hydrophilic or hydrophobic far-red emitting dyes, and exhibiting enhanced brightness. They efficiently label Hepatitis C Virus pseudotyped particles (HCVpp), more stably and reproducibly than commercial probes, and a strong fluorescence signal is observed with a high contrast. Labeling with such probes do not alter virion morphology, integrity, nor infectivity. Finally, it is shown by fluorescence microscopy that these probes enable efficient tracking of labeled HCVpp inside hepatocarcinoma cells used as model hepatocytes, in spite of their autofluorescence up to 700 nm. These novel fluorescent lipid-polymer probes should therefore enable a better characterization of early stages of infection of autofluorescent cells by enveloped viruses. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chromophore Isomer Stabilization Is Critical to the Efficient Fluorescence of Cyan Fluorescent Proteins.

PubMed

Gotthard, Guillaume; von Stetten, David; Clavel, Damien; Noirclerc-Savoye, Marjolaine; Royant, Antoine

2017-12-12

ECFP, the first usable cyan fluorescent protein (CFP), was obtained by adapting the tyrosine-based chromophore environment in green fluorescent protein to that of a tryptophan-based one. This first-generation CFP was superseded by the popular Cerulean, CyPet, and SCFP3A that were engineered by rational and random mutagenesis, yet the latter CFPs still exhibit suboptimal properties of pH sensitivity and reversible photobleaching behavior. These flaws were serendipitously corrected in the third-generation CFP mTurquoise and its successors without an obvious rationale. We show here that the evolution process had unexpectedly remodeled the chromophore environment in second-generation CFPs so they would accommodate a different isomer, whose formation is favored by acidic pH or light irradiation and which emits fluorescence much less efficiently. Our results illustrate how fluorescent protein engineering based solely on fluorescence efficiency optimization may affect other photophysical or physicochemical parameters and provide novel insights into the rational evolution of fluorescent proteins with a tryptophan-based chromophore.

Nanostructure and Corresponding Quenching Efficiency of Fluorescent DNA Probes.

PubMed

Guo, Wenjuan; Wei, Yanhong; Dai, Zhao; Chen, Guangping; Chu, Yuanyuan; Zhao, Yifei

2018-02-09

Based on the fluorescence resonance energy transfer (FRET) mechanism, fluorescent DNA probes were prepared with a novel DNA hairpin template method, with SiO₂ coated CdTe (CdTe/SiO₂) core/shell nanoparticles used as the fluorescence energy donors and gold (Au) nanoparticles (AuNPs) as the energy acceptors. The nanostructure and energy donor/acceptor ratio in a probe were controlled with this method. The relationship between the nanostructure of the probes and FRET efficiency (quenching efficiency) were investigated. The results indicated that when the donor/acceptor ratios were 2:1, 1:1, and 1:2; the corresponding FRET efficiencies were about 33.6%, 57.5%, and 74.2%, respectively. The detection results indicated that the fluorescent recovery efficiency of the detecting system was linear when the concentration of the target DNA was about 0.0446-2.230 nmol/L. Moreover, the probes showed good sensitivity and stability in different buffer conditions with a low detection limit of about 0.106 nmol/L.

Nanostructure and Corresponding Quenching Efficiency of Fluorescent DNA Probes

PubMed Central

Guo, Wenjuan; Wei, Yanhong; Dai, Zhao; Chen, Guangping; Chu, Yuanyuan; Zhao, Yifei

2018-01-01

Based on the fluorescence resonance energy transfer (FRET) mechanism, fluorescent DNA probes were prepared with a novel DNA hairpin template method, with SiO2 coated CdTe (CdTe/SiO2) core/shell nanoparticles used as the fluorescence energy donors and gold (Au) nanoparticles (AuNPs) as the energy acceptors. The nanostructure and energy donor/acceptor ratio in a probe were controlled with this method. The relationship between the nanostructure of the probes and FRET efficiency (quenching efficiency) were investigated. The results indicated that when the donor/acceptor ratios were 2:1, 1:1, and 1:2; the corresponding FRET efficiencies were about 33.6%, 57.5%, and 74.2%, respectively. The detection results indicated that the fluorescent recovery efficiency of the detecting system was linear when the concentration of the target DNA was about 0.0446–2.230 nmol/L. Moreover, the probes showed good sensitivity and stability in different buffer conditions with a low detection limit of about 0.106 nmol/L. PMID:29425163

High-throughput and selective solid-phase extraction of urinary catecholamines by crown ether-modified resin composite fiber.

PubMed

Chen, LiQin; Wang, Hui; Xu, Zhen; Zhang, QiuYue; Liu, Jia; Shen, Jun; Zhang, WanQi

2018-08-03

In the present study, we developed a simple and high-throughput solid phase extraction (SPE) procedure for selective extraction of catecholamines (CAs) in urine samples. The SPE adsorbents were electrospun composite fibers functionalized with 4-carboxybenzo-18-crown-6 ether modified XAD resin and polystyrene, which were packed into 96-well columns and used for high-throughput selective extraction of CAs in healthy human urine samples. Moreover, the extraction efficiency of packed-fiber SPE (PFSPE) was examined by high performance liquid chromatography coupled with fluorescence detector. The parameters affecting the extraction efficiency and impurity removal efficiency were optimized, and good linearity ranging from 0.5 to 400 ng/mL was obtained with a low limit of detection (LOD, 0.2-0.5 ng/mL) and a good repeatability (2.7%-3.7%, n = 6). The extraction recoveries of three CAs ranged from 70.5% to 119.5%. Furthermore, stable and reliable results obtained by the fluorescence detector were superior to those obtained by the electrochemical detector. Collectively, PFSPE coupled with 96-well columns was a simple, rapid, selective, high-throughput and cost-efficient method, and the proposed method could be applied in clinical chemistry. Copyright © 2018 Elsevier B.V. All rights reserved.

Performance of a two-leaf light use efficiency model for mapping gross primary productivity against remotely sensed sun-induced chlorophyll fluorescence data.

PubMed

Zan, Mei; Zhou, Yanlian; Ju, Weimin; Zhang, Yongguang; Zhang, Leiming; Liu, Yibo

2018-02-01

Estimating terrestrial gross primary production is an important task when studying the carbon cycle. In this study, the ability of a two-leaf light use efficiency model to simulate regional gross primary production in China was validated using satellite Global Ozone Monitoring Instrument - 2 sun-induced chlorophyll fluorescence data. The two-leaf light use efficiency model was used to estimate daily gross primary production in China's terrestrial ecosystems with 500-m resolution for the period from 2007 to 2014. Gross primary production simulated with the two-leaf light use efficiency model was resampled to a spatial resolution of 0.5° and then compared with sun-induced chlorophyll fluorescence. During the study period, sun-induced chlorophyll fluorescence and gross primary production simulated by the two-leaf light use efficiency model exhibited similar spatial and temporal patterns in China. The correlation coefficient between sun-induced chlorophyll fluorescence and monthly gross primary production simulated by the two-leaf light use efficiency model was significant (p<0.05, n=96) in 88.9% of vegetated areas in China (average value 0.78) and varied among vegetation types. The interannual variations in monthly sun-induced chlorophyll fluorescence and gross primary production simulated by the two-leaf light use efficiency model were similar in spring and autumn in most vegetated regions, but dissimilar in winter and summer. The spatial variability of sun-induced chlorophyll fluorescence and gross primary production simulated by the two-leaf light use efficiency model was similar in spring, summer, and autumn. The proportion of spatial variations of sun-induced chlorophyll fluorescence and annual gross primary production simulated by the two-leaf light use efficiency model explained by ranged from 0.76 (2011) to 0.80 (2013) during the study period. Overall, the two-leaf light use efficiency model was capable of capturing spatial and temporal variations in gross primary production in China. However, the model needs further improvement to better simulate gross primary production in summer. Copyright © 2017 Elsevier B.V. All rights reserved.

Integrated micro-optofluidic platform for real-time detection of airborne microorganisms

NASA Astrophysics Data System (ADS)

Choi, Jeongan; Kang, Miran; Jung, Jae Hee

2015-11-01

We demonstrate an integrated micro-optofluidic platform for real-time, continuous detection and quantification of airborne microorganisms. Measurements of the fluorescence and light scattering from single particles in a microfluidic channel are used to determine the total particle number concentration and the microorganism number concentration in real-time. The system performance is examined by evaluating standard particle measurements with various sample flow rates and the ratios of fluorescent to non-fluorescent particles. To apply this method to real-time detection of airborne microorganisms, airborne Escherichia coli, Bacillus subtilis, and Staphylococcus epidermidis cells were introduced into the micro-optofluidic platform via bioaerosol generation, and a liquid-type particle collection setup was used. We demonstrate successful discrimination of SYTO82-dyed fluorescent bacterial cells from other residue particles in a continuous and real-time manner. In comparison with traditional microscopy cell counting and colony culture methods, this micro-optofluidic platform is not only more accurate in terms of the detection efficiency for airborne microorganisms but it also provides additional information on the total particle number concentration.

Laser-induced fluorescence from N2(+) ions generated by a corona discharge in ambient air.

PubMed

Konthasinghe, Kumarasiri; Fitzmorris, Kristin; Peiris, Manoj; Hopkins, Adam J; Petrak, Benjamin; Killinger, Dennis K; Muller, Andreas

2015-09-01

In this work, we present the measurement of laser-induced fluorescence from N2(+) ions via the B(2)Σu(+)-X(2)Σg(+) band system in the near-ultraviolet. The ions were generated continuously by a plasma glow discharge in low pressure N2 and by a corona discharge in ambient air. The fluorescence decay time was found to rapidly decrease with increasing pressure leading to an extrapolated decay rate of ≍10(10) s(-1) at atmospheric pressure. In spite of this quenching, we were able to observe laser induced fluorescence in ambient air by means of a time-gated spectral measurement. In the process of comparing the emission signal with that of N2 spontaneous Raman scattering, ion concentrations in ambient air of order 10(8-)10(10) cm(-3) were determined. With moderate increases in laser power and collection efficiency, ion concentrations of less than 10(6) cm(-3) may be measurable, potentially enabling applications in atmospheric standoff detection of ionizing radiation from hazardous radioactive sources.

[Establishment and identification of mouse lymphoma cell line EL4 expressing red fluorescent protein].

PubMed

Li, Yan-Jie; Cao, Jiang; Chen, Chong; Wang, Dong-Yang; Zeng, Ling-Yu; Pan, Xiu-Ying; Xu, Kai-Lin

2010-02-01

This study was purposed to construct a lentiviral vector encoding red fluorescent protein (DsRed) and transfect DsRed into EL4 cells for establishing mouse leukemia/lymphoma model expressing DsRed. The bicistronic SIN lentiviral transfer plasmid containing the genes encoding neo and internal ribosomal entry site-red fluorescent protein (IRES-DsRed) was constructed. Human embryonic kidney 293FT cells were co-transfected with the three plasmids by liposome method. The viral particles were collected and used to transfect EL4 cells, then the cells were selected by G418. The results showed that the plasmid pXZ208-neo-IRES-DsRed was constructed successfully, and the viral titer reached to 10(6) U/ml. EL4 cells were transfected by the viral solution efficiently. The transfected EL4 cells expressing DsRed survived in the final concentration 600 microg/ml of G418. The expression of DsRed in the transfected EL4 cells was demonstrated by fluorescence microscopy and flow cytometry. In conclusion, the EL4/DsRed cell line was established successfully.

Integrated micro-optofluidic platform for real-time detection of airborne microorganisms

PubMed Central

Choi, Jeongan; Kang, Miran; Jung, Jae Hee

2015-01-01

We demonstrate an integrated micro-optofluidic platform for real-time, continuous detection and quantification of airborne microorganisms. Measurements of the fluorescence and light scattering from single particles in a microfluidic channel are used to determine the total particle number concentration and the microorganism number concentration in real-time. The system performance is examined by evaluating standard particle measurements with various sample flow rates and the ratios of fluorescent to non-fluorescent particles. To apply this method to real-time detection of airborne microorganisms, airborne Escherichia coli, Bacillus subtilis, and Staphylococcus epidermidis cells were introduced into the micro-optofluidic platform via bioaerosol generation, and a liquid-type particle collection setup was used. We demonstrate successful discrimination of SYTO82-dyed fluorescent bacterial cells from other residue particles in a continuous and real-time manner. In comparison with traditional microscopy cell counting and colony culture methods, this micro-optofluidic platform is not only more accurate in terms of the detection efficiency for airborne microorganisms but it also provides additional information on the total particle number concentration. PMID:26522006

Development of accelerated Raman and fluorescent Monte Carlo method

NASA Astrophysics Data System (ADS)

Dumont, Alexander P.; Patil, Chetan

2018-02-01

Monte Carlo (MC) modeling of photon propagation in turbid media is an essential tool for understanding optical interactions between light and tissue. Insight gathered from outputs of MC models assists in mapping between detected optical signals and bulk tissue optical properties, and as such, has proven useful for inverse calculations of tissue composition and optimization of the design of optical probes. MC models of Raman scattering have previously been implemented without consideration to background autofluorescence, despite its presence in raw measurements. Modeling both Raman and fluorescence profiles at high spectral resolution requires a significant increase in computation, but is more appropriate for investigating issues such as detection limits. We present a new Raman Fluorescence MC model developed atop an existing GPU parallelized MC framework that can run more than 300x times faster than CPU methods. The robust acceleration allows for the efficient production of both Raman and fluorescence outputs from the MC model. In addition, this model can handle arbitrary sample morphologies of excitation and collection geometries to more appropriately mimic experimental settings. We will present the model framework and initial results.

Comparison of femtosecond- and nanosecond-two-photon-absorption laser-induced fluorescence (TALIF) of atomic oxygen in atmospheric-pressure plasmas

NASA Astrophysics Data System (ADS)

Schmidt, Jacob B.; Sands, Brian; Scofield, James; Gord, James R.; Roy, Sukesh

2017-05-01

Absolute number densities of atomic species produced by nanosecond (ns)-duration, repetitively pulsed electric discharges are measured by two-photon-absorption laser-induced fluorescence (TALIF). Unique to this work is the development of femtosecond-laser-based TALIF (fs-TALIF) that offers a number of advantages over more conventional nanosecond (ns)-pulse-duration laser techniques, such as higher-fidelity quenching rate measurements over a wide pressure range, significantly reduced photolytic interference (including photo-dissociation and photo-ionization), ability to collect two-dimensional images of atomic-species number densities with high spatial resolution aided by higher signal level, and efficient and accurate measurements of atomic-species number densities due to the higher repetition rates of the laser. For full quantification of these advantages, atomic-oxygen TALIF signals are collected from an atmospheric-pressure plasma jet employing both ns- and fs-duration laser-excitation pulses and the results are compared and contrasted.

High-performance liquid chromatography with fast-scanning fluorescence detection and multivariate curve resolution for the efficient determination of galantamine and its main metabolites in serum.

PubMed

Culzoni, María J; Aucelio, Ricardo Q; Escandar, Graciela M

2012-08-31

Based on green analytical chemistry principles, an efficient approach was applied for the simultaneous determination of galantamine, a widely used cholinesterase inhibitor for the treatment of Alzheimer's disease, and its major metabolites in serum samples. After a simple serum deproteinization step, second-order data were rapidly obtained (less than 6 min) with a chromatographic system operating in the isocratic regime using ammonium acetate/acetonitrile (94:6) as mobile phase. Detection was made with a fast-scanning spectrofluorimeter, which allowed the efficient collection of data to obtain matrices of fluorescence intensity as a function of retention time and emission wavelength. Successful resolution was achieved in the presence of matrix interferences in serum samples using multivariate curve resolution-alternating least-squares (MCR-ALS). The developed approach allows the quantification of the analytes at levels found in treated patients, without the need of applying either preconcentration or extraction steps. Limits of detection in the range between 8 and 11 ng mL(-1), relative prediction errors from 7 to 12% and coefficients of variation from 4 to 7% were achieved. Copyright © 2012 Elsevier B.V. All rights reserved.

High-efficiency in situ resonant inelastic x-ray scattering (iRIXS) endstation at the Advanced Light Source

NASA Astrophysics Data System (ADS)

Qiao, Ruimin; Li, Qinghao; Zhuo, Zengqing; Sallis, Shawn; Fuchs, Oliver; Blum, Monika; Weinhardt, Lothar; Heske, Clemens; Pepper, John; Jones, Michael; Brown, Adam; Spucces, Adrian; Chow, Ken; Smith, Brian; Glans, Per-Anders; Chen, Yanxue; Yan, Shishen; Pan, Feng; Piper, Louis F. J.; Denlinger, Jonathan; Guo, Jinghua; Hussain, Zahid; Chuang, Yi-De; Yang, Wanli

2017-03-01

An endstation with two high-efficiency soft x-ray spectrographs was developed at Beamline 8.0.1 of the Advanced Light Source, Lawrence Berkeley National Laboratory. The endstation is capable of performing soft x-ray absorption spectroscopy, emission spectroscopy, and, in particular, resonant inelastic soft x-ray scattering (RIXS). Two slit-less variable line-spacing grating spectrographs are installed at different detection geometries. The endstation covers the photon energy range from 80 to 1500 eV. For studying transition-metal oxides, the large detection energy window allows a simultaneous collection of x-ray emission spectra with energies ranging from the O K-edge to the Ni L-edge without moving any mechanical components. The record-high efficiency enables the recording of comprehensive two-dimensional RIXS maps with good statistics within a short acquisition time. By virtue of the large energy window and high throughput of the spectrographs, partial fluorescence yield and inverse partial fluorescence yield signals could be obtained for all transition metal L-edges including Mn. Moreover, the different geometries of these two spectrographs (parallel and perpendicular to the horizontal polarization of the beamline) provide contrasts in RIXS features with two different momentum transfers.

Amyloid-β Deposits Target Efficient Near-Infrared Fluorescent Probes: Synthesis, in Vitro Evaluation, and in Vivo Imaging.

PubMed

Fu, Hualong; Tu, Peiyu; Zhao, Liu; Dai, Jiapei; Liu, Boli; Cui, Mengchao

2016-02-02

The formation of extracellular amyloid-β (Aβ) plaques is a common molecular change that underlies several debilitating human conditions, including Alzheimer's disease (AD); however, the existing near-infrared (NIR) fluorescent probes for the in vivo detection of Aβ plaques are limited by undesirable fluorescent properties and poor brain kinetics. In this work, we designed, synthesized, and evaluated a new family of efficient NIR probes that target Aβ plaques by incorporating hydroxyethyl groups into the ligand structure. Among these probes, DANIR 8c showed excellent fluorescent properties with an emission maximum above 670 nm upon binding to Aβ aggregates and also displayed a high sensitivity (a 629-fold increase in fluorescence intensity) and affinity (Kd = 14.5 nM). Because of the improved hydrophilicity that was induced by hydroxyls, 8c displayed increased initial brain uptake and a fast washout from the brain, as well as an acceptable biostability in the brain. In vivo NIR fluorescent imaging revealed that 8c could efficiently distinguish between AD transgenic model mice and normal controls. Overall, 8c is an efficient and veritable NIR fluorescent probe for the in vivo detection of Aβ plaques in the brain.

A novel multiwavelength fluorescence image-guided surgery imaging system

NASA Astrophysics Data System (ADS)

Volpi, D.; Tullis, I. D. C.; Laios, A.; Pathiraja, P. N. J.; Haldar, K.; Ahmed, A. A.; Vojnovic, B.

2014-02-01

We describe the development and performance analysis of two clinical near-infrared fluorescence image-guided surgery (FIGS) devices that aim to overcome some of the limitations of current FIGS systems. The devices operate in a widefield-imaging mode and can work (1) in conjunction with a laparoscope, during minimally invasive surgery, and (2) as a hand-held, open surgery imaging system. In both cases, narrow-band excitation light, delivered at multiple wavelengths, is efficiently combined with white reflectance light. Light is delivered to ~100 cm2 surgical field at 1-2 mW/cm2 for white light and 3-7 mW/cm2 (depending on wavelength) of red - near infrared excitation, at a typical working distance of 350 mm for the hand-held device and 100 mm for the laparoscope. A single, sensitive, miniaturized color camera collects both fluorescence and white reflectance light. The use of a single imager eliminates image alignment and software overlay complexity. A novel filtering and illumination arrangement allows simultaneous detection of white reflectance and fluorescence emission from multiple dyes in real-time. We will present both fluorescence detection sensitivity modeling and practical performance data. We have demonstrated the efficiency and the advantages of the devices both pre-clinically and during live surgery on humans. Both the hand-held and the laparoscopic systems have proved to be reliable and beneficial in an ongoing clinical trial involving sentinel lymph node detection in gynecological cancers. We will show preliminary results using two clinically approved dyes, Methylene blue and indocyanine green. We anticipate that this technology can be integrated and routinely used in a larger variety of surgical procedures.

Nanosphere-based one-step strategy for efficient and nondestructive detection of circulating tumor cells.

PubMed

Wu, Ling-Ling; Wen, Cong-Ying; Hu, Jiao; Tang, Man; Qi, Chu-Bo; Li, Na; Liu, Cui; Chen, Lan; Pang, Dai-Wen; Zhang, Zhi-Ling

2017-08-15

Detecting viable circulating tumor cells (CTCs) without disruption to their functions for in vitro culture and functional study could unravel the biology of metastasis and promote the development of personalized anti-tumor therapies. However, existing CTC detection approaches commonly include CTC isolation and subsequent destructive identification, which damages CTC viability and functions and generates substantial CTC loss. To address the challenge of efficiently detecting viable CTCs for functional study, we develop a nanosphere-based cell-friendly one-step strategy. Immunonanospheres with prominent magnetic/fluorescence properties and extraordinary stability in complex matrices enable simultaneous efficient magnetic capture and specific fluorescence labeling of tumor cells directly in whole blood. The collected cells with fluorescent tags can be reliably identified, free of the tedious and destructive manipulations from conventional CTC identification. Hence, as few as 5 tumor cells in ca. 1mL of whole blood can be efficiently detected via only 20min incubation, and this strategy also shows good reproducibility with the relative standard deviation (RSD) of 8.7%. Moreover, due to the time-saving and gentle processing and the minimum disruption of immunonanospheres to cells, 93.8±0.1% of detected tumor cells retain cell viability and proliferation ability with negligible changes of cell functions, capacitating functional study on cell migration, invasion and glucose uptake. Additionally, this strategy exhibits successful CTC detection in 10/10 peripheral blood samples of cancer patients. Therefore, this nanosphere-based cell-friendly one-step strategy enables viable CTC detection and further functional analyses, which will help to unravel tumor metastasis and guide treatment selection. Copyright © 2017 Elsevier B.V. All rights reserved.

Phytoplankton-Fluorescence-Lifetime Vertical Profiler

NASA Technical Reports Server (NTRS)

Fernandez, Salvador M.; Guignon, Ernest F.; St. Louis, Ernest

2004-01-01

A battery-operated optoelectronic instrument is designed to be lowered into the ocean to measure the intensity and lifetime of fluorescence of chlorophyll A in marine phytoplankton as a function of depth from 0 to 300 m. Fluorescence lifetimes are especially useful as robust measures of photosynthetic productivity of phytoplankton and of physical and chemical mechanisms that affect photosynthesis. The knowledge of photosynthesis in phytoplankton gained by use of this and related instruments is expected to contribute to understanding of global processes that control the time-varying fluxes of carbon and associated biogenic elements in the ocean. The concentration of chlorophyll in the ocean presents a major detection challenge because in order to obtain accurate values of photosynthetic parameters, the intensity of light used to excite fluorescence must be kept very low so as not to disturb the photosynthetic system. Several innovations in fluorometric instrumentation were made in order to make it possible to reach the required low detection limit. These innovations include a highly efficient optical assembly with an integrated flow-through sample interface, and a high-gain, low-noise electronic detection subsystem. The instrument also incorporates means for self-calibration during operation, and electronic hardware and software for control, acquisition and analysis of data, and communications. The electronic circuitry is highly miniaturized and designed to minimize power demand. The instrument is housed in a package that can withstand the water pressure at the maximum depth of 300 m. A light-emitting diode excites fluorescence in the sample flow cell, which is placed at one focal point of an ellipsoidal reflector. A photomultiplier tube is placed at the other focal point. This optical arrangement enables highly efficient collection of fluorescence emitted over all polar directions. Fluorescence lifetime is measured indirectly, by use of a technique based on the same principle as the one described in "Fluorometer for Analysis of Photosynthesis in Phytoplankton" (SSC-00110), NASA Tech Briefs, Vol. 24, No. 1 (November 2000), page 79. The excitation is modulated at a frequency of 70 MHz, and the phase shift between the excitation light and the emitted fluorescence is measured by a detection method in which the 70 MHz signal is down-converted to a 400 Hz signal. The fluorescence lifetime can be computed from the known relationship among the fluorescence lifetime, phase shift, and modulation frequency

A cylindrical quadrupole ion trap in combination with an electrospray ion source for gas-phase luminescence and absorption spectroscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stockett, Mark H., E-mail: stockett@phys.au.dk; Houmøller, Jørgen; Støchkel, Kristian

2016-05-15

A relatively simple setup for collection and detection of light emitted from isolated photo-excited molecular ions has been constructed. It benefits from a high collection efficiency of photons, which is accomplished by using a cylindrical ion trap where one end-cap electrode is a mesh grid combined with an aspheric condenser lens. The geometry permits nearly 10% of the emitted light to be collected and, after transmission losses, approximately 5% to be delivered to the entrance of a grating spectrometer equipped with a detector array. The high collection efficiency enables the use of pulsed tunable lasers with low repetition rates (e.g.,more » 20 Hz) instead of continuous wave (cw) lasers or very high repetition rate (e.g., MHz) lasers that are typically used as light sources for gas-phase fluorescence experiments on molecular ions. A hole has been drilled in the cylinder electrode so that a light pulse can interact with the ion cloud in the center of the trap. Simulations indicate that these modifications to the trap do not significantly affect the storage capability and the overall shape of the ion cloud. The overlap between the ion cloud and the laser light is basically 100%, and experimentally >50% of negatively charged chromophore ions are routinely photodepleted. The performance of the setup is illustrated based on fluorescence spectra of several laser dyes, and the quality of these spectra is comparable to those reported by other groups. Finally, by replacing the optical system with a channeltron detector, we demonstrate that the setup can also be used for gas-phase action spectroscopy where either depletion or fragmentation is monitored to provide an indirect measurement on the absorption spectrum of the ion.« less

A cylindrical quadrupole ion trap in combination with an electrospray ion source for gas-phase luminescence and absorption spectroscopy.

PubMed

Stockett, Mark H; Houmøller, Jørgen; Støchkel, Kristian; Svendsen, Annette; Brøndsted Nielsen, Steen

2016-05-01

A relatively simple setup for collection and detection of light emitted from isolated photo-excited molecular ions has been constructed. It benefits from a high collection efficiency of photons, which is accomplished by using a cylindrical ion trap where one end-cap electrode is a mesh grid combined with an aspheric condenser lens. The geometry permits nearly 10% of the emitted light to be collected and, after transmission losses, approximately 5% to be delivered to the entrance of a grating spectrometer equipped with a detector array. The high collection efficiency enables the use of pulsed tunable lasers with low repetition rates (e.g., 20 Hz) instead of continuous wave (cw) lasers or very high repetition rate (e.g., MHz) lasers that are typically used as light sources for gas-phase fluorescence experiments on molecular ions. A hole has been drilled in the cylinder electrode so that a light pulse can interact with the ion cloud in the center of the trap. Simulations indicate that these modifications to the trap do not significantly affect the storage capability and the overall shape of the ion cloud. The overlap between the ion cloud and the laser light is basically 100%, and experimentally >50% of negatively charged chromophore ions are routinely photodepleted. The performance of the setup is illustrated based on fluorescence spectra of several laser dyes, and the quality of these spectra is comparable to those reported by other groups. Finally, by replacing the optical system with a channeltron detector, we demonstrate that the setup can also be used for gas-phase action spectroscopy where either depletion or fragmentation is monitored to provide an indirect measurement on the absorption spectrum of the ion.

Carbon "Quantum" Dots for Fluorescence Labeling of Cells.

PubMed

Liu, Jia-Hui; Cao, Li; LeCroy, Gregory E; Wang, Ping; Meziani, Mohammed J; Dong, Yiyang; Liu, Yuanfang; Luo, Pengju G; Sun, Ya-Ping

2015-09-02

The specifically synthesized and selected carbon dots of relatively high fluorescence quantum yields were evaluated in their fluorescence labeling of cells. For the cancer cell lines, the cellular uptake of the carbon dots was generally efficient, resulting in the labeling of the cells with bright fluorescence emissions for both one- and two-photon excitations from predominantly the cell membrane and cytoplasm. In the exploration on labeling the live stem cells, the cellular uptake of the carbon dots was relatively less efficient, though fluorescence emissions could still be adequately detected in the labeled cells, with the emissions again predominantly from the cell membrane and cytoplasm. This combined with the observed more efficient internalization of the same carbon dots by the fixed stem cells might suggest some significant selectivity of the stem cells toward surface functionalities of the carbon dots. The needs and possible strategies for more systematic and comparative studies on the fluorescence labeling of different cells, including especially live stem cells, by carbon dots as a new class of brightly fluorescent probes are discussed.

Photo-acoustic spectroscopy and quantum efficiency of Yb{sup 3+} doped alumino silicate glasses

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuhn, Stefan, E-mail: stefan.kuhn84@googlemail.com; Tiegel, Mirko; Herrmann, Andreas

2015-09-14

In this contribution, we analyze the effect of several preparation methods of Yb{sup 3+} doped alumino silicate glasses on their quantum efficiency by using photo-acoustic measurements in comparison to standard measurement methods including the determination via the fluorescence lifetime and an integrating sphere setup. The preparation methods focused on decreasing the OH concentration by means of fluorine-substitution and/or applying dry melting atmospheres, which led to an increase in the measured fluorescence lifetime. However, it was found that the influence of these methods on radiative properties such as the measured fluorescence lifetime alone does not per se give exact information aboutmore » the actual quantum efficiency of the sample. The determination of the quantum efficiency by means of fluorescence lifetime shows inaccuracies when refractive index changing elements such as fluorine are incorporated into the glass. Since fluorine not only eliminates OH from the glass but also increases the “intrinsic” radiative fluorescence lifetime, which is needed to calculate the quantum efficiency, it is difficult to separate lifetime quenching from purely radiative effects. The approach used in this contribution offers a possibility to disentangle radiative from non-radiative properties which is not possible by using fluorescence lifetime measurements alone and allows an accurate determination of the quantum efficiency of a given sample. The comparative determination by an integrating sphere setup leads to the well-known problem of reabsorption which embodies itself in the measurement of too low quantum efficiencies, especially for samples with small quantum efficiencies.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stuckelberger, Michael; Nietzold, Tara; Hall, Genevieve N.

Unveiling the correlation between elemental composition, Fermi-level splitting, and charge collection in perovskite solar cells (PSCs) when exposed to different environments is crucial to understanding the origin of defects. This will enable defect engineering to achieve high-performance and long-lasting PSCs. Here, in this paper, we measured, for the first time, the spatial distribution and charge-collection efficiency at the nanoscale by synchrotron-based X-ray fluorescence (XRF) and X-ray beam-induced current (XBIC) with subgrain resolution, and we observe a correlation between Pb/I ratio and charge-collection efficiency. In contrast with other thin-film solar cells, PSCs are highly sensitive to ambient conditions (atmosphere and illumination).more » As the XRF and XBIC measurements were conducted in vacuum under an X-ray source illumination, the impact of measurement conditions on the cells needs to be taken into account. Furthermore, necessary conditions for quantification of XRF/XBIC measurements, such as film homogeneity, are not fulfilled in the case of PSCs. Finally, we will discuss fundamentals of XRF/XBIC measurements of PSCs that will enable reliable, quantitative, high-resolution measurements of elemental distribution and charge collection.« less

Photochemical studies of a fluorescent chlorophyll catabolite--source of bright blue fluorescence in plant tissue and efficient sensitizer of singlet oxygen.

PubMed

Jockusch, Steffen; Turro, Nicholas J; Banala, Srinivas; Kräutler, Bernhard

2014-02-01

Fluorescent chlorophyll catabolites (FCCs) are fleeting intermediates of chlorophyll breakdown, which is seen as an enzyme controlled detoxification process of the chlorophylls in plants. However, some plants accumulate large amounts of persistent FCCs, such as in senescent leaves and in peels of yellow bananas. The photophysical properties of such a persistent FCC (Me-sFCC) were investigated in detail. FCCs absorb in the near UV spectral region and show blue fluorescence (max at 437 nm). The Me-sFCC fluorescence had a quantum yield of 0.21 (lifetime 1.6 ns). Photoexcited Me-sFCC intersystem crosses into the triplet state (quantum yield 0.6) and generates efficiently singlet oxygen (quantum yield 0.59). The efficient generation of singlet oxygen makes fluorescent chlorophyll catabolites phototoxic, but might also be useful as a (stress) signal and for defense of the plant tissue against infection by pathogens.

An Experimental Study of the Fluorescence Spectrum of Cesium Atoms in the Presence of a Buffer Gas

NASA Astrophysics Data System (ADS)

Davydov, V. G.; Kulyasov, V. N.

2018-01-01

A direct experiment is performed to determine the quantum efficiency of a cesium fluorescence filter. The fluorescence spectra of cesium atoms are recorded under excitation of the upper states of the second resonance doublet with a Bell-Bloom cesium lamp. Introduction of different noble gases into the cell with cesium leads to the appearance of additional fluorescence photons. It is found that a fluorescence filter based on atomic cesium vapor with addition of helium in the working cell has the highest efficiency and response rate of all known fluorescence filters based on alkali-metal atomic vapors.

The 2013 FLEX-US Airborne Campaign at the Parker Tract Loblolly Pine Plantation in North Carolina, USA

NASA Technical Reports Server (NTRS)

Middleton, Elizabeth M.; Rascher, Uwe; Corp, Lawrence A.; Huemmrich, K. Fred; Cook, Bruce D.; Noormets, Asko; Schickling, Anke; Pinto, Francisco; Alonso, Luis; Damm, Alexander;

Thioflavin T as an efficient fluorescence sensor for selective recognition of RNA G-quadruplexes

NASA Astrophysics Data System (ADS)

Xu, Shujuan; Li, Qian; Xiang, Junfeng; Yang, Qianfan; Sun, Hongxia; Guan, Aijiao; Wang, Lixia; Liu, Yan; Yu, Lijia; Shi, Yunhua; Chen, Hongbo; Tang, Yalin

2016-04-01

RNA G-quadruplexes (G4s) play important roles in translational regulation, mRNA processing events and gene expression. Therefore, a fluorescent probe that is capable of efficiently recognizing RNA G-quadruplex structures among other RNA forms is highly desirable. In this study, a water-soluble fluorogenic dye (i.e., Thioflavin T (ThT)) was employed to recognize RNA G-quadruplex structures using UV-Vis absorption spectra, fluorescence spectra and emission lifetime experiments. By stacking on the G-tetrad, the ThT probe exhibited highly specific recognition of RNA G-quadruplex structures with striking fluorescence enhancement compared with other RNA forms. The specific binding demonstrates that ThT is an efficient fluorescence sensor that can distinguish G4 and non-G4 RNA structures.

Maximizing Aggregation of Organic Fluorophores to Prolong Fluorescence Lifetime for Two-Photon Fluorescence Lifetime Imaging.

PubMed

Hu, Wenbo; Guo, Lihong; Bai, Lei; Miao, Xiaofei; Ni, Yun; Wang, Qi; Zhao, Hui; Xie, Meng; Li, Lin; Lu, Xiaomei; Huang, Wei; Fan, Quli

2018-05-28

Two-photon fluorescence lifetime imaging (TP-FLIM) not only permits imaging deep inside the tissues with precise spatial manipulation but also circumvents tissue autofluorescence, holding tremendous promise in molecular imaging. However, the serious lack of suitable contrast agents with long fluorescence lifetime and efficient two-photon absorption (TPA) greatly limits the advance of TP-FLIM. This study reports a simple approach to fabricate water-soluble organic semiconducting nanoparticles [thioxanthone (TXO) NPs] with ultralong fluorescence lifetime and efficient TPA for in vivo TP-FLIM. The approach utilizes the aggregation of a specifically selected thermally activated delayed fluorescence (TADF) fluorophore to prolong its fluorescence lifetime. Encapsulating the TADF fluorophore within an amphiphilic copolymer not only maximizes its aggregation but also obtains TXO NPs with efficient TPA. Importantly, as-prepared TXO NPs exhibit a considerably long fluorescence lifetime at a magnitude of 4.2 µs, which is almost 1000 times larger than that of existing organic contrast agents. Moreover, such long fluorescence lifetime is almost oxygen-inert, readily realizing both in vitro and in vivo TP-FLIM. This work may set valuable guidance for designing organic semiconducting materials with ultralong fluorescence lifetimes to fulfill the potential of FLIM. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Nd/sup 3 +/ fluorescence quantum-efficiency measurements with photoacoustics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rosencwaig, A.; Hildum, E.A.

1981-04-01

We have investigated the use of photoacoustic techniques for obtaining absolute values of fluorescence quantum efficiencies in lightly doped Nd/sup 3 +/ laser materials. We have found that surface absorptions play an important role in gas-microphone measurements, and that thermal profiles are important in piezoelectric measurements. We have obtained fluorescence quantum efficiencies for Nd/sup 3 +/ in yttrium aluminum garnet, and in silicate and borate glasses that are in good agreement with lifetime measurements and Judd-Ofelt calculations.

New Monte Carlo model of cylindrical diffusing fibers illustrates axially heterogeneous fluorescence detection: simulation and experimental validation

PubMed Central

Baran, Timothy M.; Foster, Thomas H.

2011-01-01

We present a new Monte Carlo model of cylindrical diffusing fibers that is implemented with a graphics processing unit. Unlike previously published models that approximate the diffuser as a linear array of point sources, this model is based on the construction of these fibers. This allows for accurate determination of fluence distributions and modeling of fluorescence generation and collection. We demonstrate that our model generates fluence profiles similar to a linear array of point sources, but reveals axially heterogeneous fluorescence detection. With axially homogeneous excitation fluence, approximately 90% of detected fluorescence is collected by the proximal third of the diffuser for μs'/μa = 8 in the tissue and 70 to 88% is collected in this region for μs'/μa = 80. Increased fluorescence detection by the distal end of the diffuser relative to the center section is also demonstrated. Validation of these results was performed by creating phantoms consisting of layered fluorescent regions. Diffusers were inserted into these layered phantoms and fluorescence spectra were collected. Fits to these spectra show quantitative agreement between simulated fluorescence collection sensitivities and experimental results. These results will be applicable to the use of diffusers as detectors for dosimetry in interstitial photodynamic therapy. PMID:21895311

Charge Collection in Hybrid Perovskite Solar Cells: Relation to the Nanoscale Elemental Distribution

DOE PAGES

Stuckelberger, Michael; Nietzold, Tara; Hall, Genevieve N.; ...

2016-12-19

Unveiling the correlation between elemental composition, Fermi-level splitting, and charge collection in perovskite solar cells (PSCs) when exposed to different environments is crucial to understanding the origin of defects. This will enable defect engineering to achieve high-performance and long-lasting PSCs. Here, in this paper, we measured, for the first time, the spatial distribution and charge-collection efficiency at the nanoscale by synchrotron-based X-ray fluorescence (XRF) and X-ray beam-induced current (XBIC) with subgrain resolution, and we observe a correlation between Pb/I ratio and charge-collection efficiency. In contrast with other thin-film solar cells, PSCs are highly sensitive to ambient conditions (atmosphere and illumination).more » As the XRF and XBIC measurements were conducted in vacuum under an X-ray source illumination, the impact of measurement conditions on the cells needs to be taken into account. Furthermore, necessary conditions for quantification of XRF/XBIC measurements, such as film homogeneity, are not fulfilled in the case of PSCs. Finally, we will discuss fundamentals of XRF/XBIC measurements of PSCs that will enable reliable, quantitative, high-resolution measurements of elemental distribution and charge collection.« less

Strong antenna-enhanced fluorescence of a single light-harvesting complex shows photon antibunching

PubMed Central

Wientjes, Emilie; Renger, Jan; Curto, Alberto G.; Cogdell, Richard; van Hulst, Niek F.

2014-01-01

The nature of the highly efficient energy transfer in photosynthetic light-harvesting complexes is a subject of intense research. Unfortunately, the low fluorescence efficiency and limited photostability hampers the study of individual light-harvesting complexes at ambient conditions. Here we demonstrate an over 500-fold fluorescence enhancement of light-harvesting complex 2 (LH2) at the single-molecule level by coupling to a gold nanoantenna. The resonant antenna produces an excitation enhancement of circa 100 times and a fluorescence lifetime shortening to ~\

Strong antenna-enhanced fluorescence of a single light-harvesting complex shows photon antibunching

NASA Astrophysics Data System (ADS)

Wientjes, Emilie; Renger, Jan; Curto, Alberto G.; Cogdell, Richard; van Hulst, Niek F.

2014-06-01

The nature of the highly efficient energy transfer in photosynthetic light-harvesting complexes is a subject of intense research. Unfortunately, the low fluorescence efficiency and limited photostability hampers the study of individual light-harvesting complexes at ambient conditions. Here we demonstrate an over 500-fold fluorescence enhancement of light-harvesting complex 2 (LH2) at the single-molecule level by coupling to a gold nanoantenna. The resonant antenna produces an excitation enhancement of circa 100 times and a fluorescence lifetime shortening to ~\

Development of Highly Fluorescent Materials Based on Thiophenylimidazole Dyes

NASA Technical Reports Server (NTRS)

Santos, Javier; Bu, Xiu R.; Mintz, Eric A.; Meador, Michael A. (Technical Monitor)

2000-01-01

Organic fluorescent materials are expected to find many potential applications in optical devices and photo-functionalized materials. Although many investigations have been focused on heterocyclic compounds such as coumarins, bipyridines, rhodamines, and pyrrole derivatives, little is known for fluorescent imidazole materials. We discovered that one particular class of imidazole derivatives is highly fluorescent. A series of monomeric and polymeric based fluorescent dyes were prepared containing a thiophene unit at the second position of the imidazole ring. Dependence of fluorescence efficiency on parameters such as solvent polarity and substituent groups has been investigated. It was found that a formyl group at the 2-position of the thiophene ring dramatically enhance fluorescence properties. Ion recognition probes indicated their potential as sensor materials. These fluorophores have flexibility for introduction of versatile substituent groups that could improve the fluorescence efficiency and sensor properties.

Classic maximum entropy recovery of the average joint distribution of apparent FRET efficiency and fluorescence photons for single-molecule burst measurements.

PubMed

DeVore, Matthew S; Gull, Stephen F; Johnson, Carey K

2012-04-05

We describe a method for analysis of single-molecule Förster resonance energy transfer (FRET) burst measurements using classic maximum entropy. Classic maximum entropy determines the Bayesian inference for the joint probability describing the total fluorescence photons and the apparent FRET efficiency. The method was tested with simulated data and then with DNA labeled with fluorescent dyes. The most probable joint distribution can be marginalized to obtain both the overall distribution of fluorescence photons and the apparent FRET efficiency distribution. This method proves to be ideal for determining the distance distribution of FRET-labeled biomolecules, and it successfully predicts the shape of the recovered distributions.

Recognition of iron ions by carbazole-desferrioxamine fluorescent sensor and its application in total iron detection in airbone particulate matter.

PubMed

Delattre, François; Cazier-Dennin, Francine; Leleu, Ludovic; Dewaele, Dorothée; Landy, David; Mallard, Isabelle; Danjou, Pierre-Edouard

2015-11-01

This work reports on an efficient microwave irradiation synthesis of a new fluorescent chemosensor based on desferrioxamine B (DFO-B) and carbazole moiety. Furthermore, this novel chemosensor was employed for a comparative study of real environmental samples of airbone particulate matter collected from Dunkirk (Northern of France). Among selected relevant metal cations present in its airbone particulate matter, such as Na(+), K(+), Mg(2+), Ca(2+), Al(3+), Cr(3+), Mn(2+) and Zn(2+), this molecular device proved to be outstandingly sensitive toward Fe(3+) with a limit of detection of 1.49 ppb (2.1×10(-8) M) in methanol allowing the estimation of total iron in atmospheric particles. Copyright © 2015 Elsevier B.V. All rights reserved.

Determination of the termination efficiency of the transcription terminator using different fluorescent profiles in green fluorescent protein mutants.

PubMed

Nojima, Takahiko; Lin, Angela C; Fujii, Teruo; Endo, Isao

2005-12-01

An approach in determining the intrinsic termination efficiency (%T) of transcription termination using green fluorescent protein (GFP) mutants was developed. This approach utilizes a cassette vector in which the tested terminator is introduced between two GFP mutant genes: an ultraviolet-optimized mutant (GFPuv: F99S, M153T, V163A) and a blue-shifted mutant (BFP: F64L, S65T, T145F). The ratio of the fluorescence intensity of BFP to GFPuv after transcription and translation represents the termination efficiency of the terminator. E. coli ribosomal RNA operon T1 terminator, phage lambda terminator site R2, E. coli tryptophane attenuater were introduced into the vector, and their transcriptional efficiencies were estimated as 89, 79, and 24%, respectively, showing good agreement with published data.

Blocking Energy-Loss Pathways for Ideal Fluorescent Organic Light-Emitting Diodes with Thermally Activated Delayed Fluorescent Sensitizers.

PubMed

Zhang, Dongdong; Song, Xiaozeng; Cai, Minghan; Duan, Lian

2018-02-01

Organic light-emitting diodes (OLEDs) based on thermally activated delayed fluorescence-sensitized fluorescence (TSF) offer the possibility of attaining an ultimate high efficiency with low roll-off utilizing noble-metal free, easy-to-synthesize, pure organic fluorescent emitters. However, the performances of TSF-OLEDs are still unsatisfactory. Here, TSF-OLEDs with breakthrough efficiencies even at high brightnesses by suppressing the competitive deactivation processes, including direct charge recombination on conventional fluorescent dopants (CFDs) and Dexter energy transfer from the host to the CFDs, are demonstrated. On the one hand, electronically inert terminal-substituents are introduced to protect the electronically active core of the CFDs; on the other hand, delicate device structures are designed to provide multiple energy-funneling paths. As a result, unprecedentedly high maximum external quantum efficiency/power efficiency of 24%/71.4 lm W -1 in a green TSF-OLED are demonstrated, which remain at 22.6%/52.3 lm W -1 even at a high luminance of 5000 cd m -2 . The work unlocks the potential of TSF-OLEDs, paving the way toward practical applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Highly efficient red OLEDs using DCJTB as the dopant and delayed fluorescent exciplex as the host.

PubMed

Zhao, Bo; Zhang, Tianyou; Chu, Bei; Li, Wenlian; Su, Zisheng; Wu, Hairuo; Yan, Xingwu; Jin, Fangming; Gao, Yuan; Liu, Chengyuan

2015-05-29

In this manuscript, we demonstrated a highly efficient DCJTB emission with delayed fluorescent exciplex TCTA:3P-T2T as the host. For the 1.0% DCJTB doped concentration, a maximum luminance, current efficiency, power efficiency and EQE of 22,767 cd m(-2), 22.7 cd A(-1), 21.5 lm W(-1) and 10.15% were achieved, respectively. The device performance is the best compared to either red OLEDs with traditional fluorescent emitter or traditional red phosphor of Ir(piq)3 doped into CBP host. The extraction of so high efficiency can be explained as the efficient triplet excitons up-conversion of TCTA:3P-T2T and the energy transfer from exciplex host singlet state to DCJTB singlet state.

Plane wave analysis of coherent holographic image reconstruction by phase transfer (CHIRPT).

PubMed

Field, Jeffrey J; Winters, David G; Bartels, Randy A

2015-11-01

Fluorescent imaging plays a critical role in a myriad of scientific endeavors, particularly in the biological sciences. Three-dimensional imaging of fluorescent intensity often requires serial data acquisition, that is, voxel-by-voxel collection of fluorescent light emitted throughout the specimen with a nonimaging single-element detector. While nonimaging fluorescence detection offers some measure of scattering robustness, the rate at which dynamic specimens can be imaged is severely limited. Other fluorescent imaging techniques utilize imaging detection to enhance collection rates. A notable example is light-sheet fluorescence microscopy, also known as selective-plane illumination microscopy, which illuminates a large region within the specimen and collects emitted fluorescent light at an angle either perpendicular or oblique to the illumination light sheet. Unfortunately, scattering of the emitted fluorescent light can cause blurring of the collected images in highly turbid biological media. We recently introduced an imaging technique called coherent holographic image reconstruction by phase transfer (CHIRPT) that combines light-sheet-like illumination with nonimaging fluorescent light detection. By combining the speed of light-sheet illumination with the scattering robustness of nonimaging detection, CHIRPT is poised to have a dramatic impact on biological imaging, particularly for in vivo preparations. Here we present the mathematical formalism for CHIRPT imaging under spatially coherent illumination and present experimental data that verifies the theoretical model.

Conformational Control of Ultrafast Molecular Rotor Property: Tuning Viscosity Sensing Efficiency by Twist Angle Variation.

PubMed

Ghosh, Rajib; Kushwaha, Archana; Das, Dipanwita

2017-09-21

Fluorescent molecular rotors find widespread application in sensing and imaging of microscopic viscosity in complex chemical and biological media. Development of viscosity-sensitive ultrafast molecular rotor (UMR) relies upon the understanding of the excited-state dynamics and their implications for viscosity-dependent fluorescence signaling. Unraveling the structure-property relationship of UMR behavior is of significance toward development of an ultrasensitive fluorescence microviscosity sensor. Herein we show that the ground-state equilibrium conformation has an important role in the ultrafast twisting dynamics of UMRs and consequent viscosity sensing efficiency. Synthesis, photophysics, and ultrafast spectroscopic experiments in conjunction with quantum chemical calculation of a series of UMRs based on dimethylaniline donor and benzimidazolium acceptor with predefined ground-state torsion angle led us to unravel that the ultrafast torsional dynamics around the bond connecting donor and acceptor groups profoundly influences the molecular rotor efficiency. This is the first experimental demonstration of conformational control of small-molecule-based UMR efficiencies which can have wider implication toward development of fluorescence sensors based on the UMR principle. Conformation-controlled UMR efficiency has been shown to exhibit commensurate fluorescence enhancement upon DNA binding.

Frequency-encoded laser-induced fluorescence for multiplexed detection in infrared-mediated quantitative PCR

PubMed Central

Schrell, Adrian M.; Roper, Michael G.

2014-01-01

A frequency-modulated fluorescence encoding method was used as a means to increase the number of fluorophores monitored during infrared-mediated polymerase chain reaction. Laser lines at 488-nm and 561-nm were modulated at 73- and 137-Hz, respectively, exciting fluorescence from the dsDNA intercalating dye, EvaGreen, and the temperature insensitive dye, ROX. Emission was collected in a color-blind manner using a single photomultiplier tube for detection and demodulated by frequency analysis. The resulting frequency domain signal resolved the contribution from the two fluorophores as well as the background from the IR lamp. The detection method was successfully used to measure amplification of DNA samples containing 104 – 107 starting copies of template producing an amplification efficiency of 96%. The utility of this methodology was further demonstrated by simultaneous amplification of two genes from human genomic DNA using different color TaqMan probes. This method of multiplexing fluorescence detection with IR-qPCR is ideally suited as it allowed isolation of the signals of interest from the background in the frequency domain and is expected to further reduce the complexity of multiplexed microfluidic IR-qPCR instrumentation. PMID:24448431

[Establishment of RAW264.7 cell strain stably expressing RFP-GFP-LC3].

PubMed

Wang, Wan; Zhang, Qing; Zhao, Runpeng; Xu, Xuewei; Xing, Yingru; Zhang, Rongbo; Wu, Jing; Hu, Dong

2015-09-01

To establish murine macrophage RAW264.7 cell strain with stable expression of red fluorescent protein-green fluorescent protein-microtubule associated protein light chain 3 (RFP-GFP-LC3). A lentiviral vector containing RFP-GFP-LC3 gene was constructed and then packaged in HEK293T cells with the packaging plasmids. The viral supernatant was collected to infect RAW264.7 cells. The RAW264.7 cell strain with stable expression of RFP-GFP-LC3 was screened with puromycin and analyzed with flow cytometry and fluorescent microscopy for infection efficiency. The number of RFP-GFP-LC3 puncta was observed using florescence microscopy following starvation treatment. The recombinant lentivirus pLV-CMV-RFP-GFP-LC3 was successfully constructed. The RAW264.7 cells with stable expression of RFP-GFP-LC3 were obtained by viral infection and puromycin screening. Fluorescent microscopy and flow cytometry demonstrated the expression rates of RFP and GFP reached to 100%. The number of autophagic puncta significantly increased after starvation treatment. The RAW264.7 cell strain with stable expression of RFP-GFP-LC3 has been successfully constructed, which provides a reliable cellular platform for autophagy research.

Improvement of antigen detection efficiency with the use of two-dimensional photonic crystal as a substrate

NASA Astrophysics Data System (ADS)

Dovzhenko, Dmitriy; Terekhin, Vladimir; Vokhmincev, Kirill; Sukhanova, Alyona; Nabiev, Igor

2017-01-01

Multiplex detection of different antigens in human serum in order to reveal diseases at the early stage is of interest nowadays. There are a lot of biosensors, which use the fluorescent labels for specific detection of analytes. For instance, common method for detection of antigens in human serum samples is enzyme-linked immunosorbent assay (ELISA). One of the most effective ways to improve the sensitivity of this detection method is the use of a substrate that could enhance the fluorescent signal and make it easier to collect. Two-dimensional (2D) photonic crystals are very suitable structures for these purposes because of the ability to enhance the luminescent signal, control the light propagation and perform the analysis directly on its surface. In our study we have calculated optimal parameters for 2D-dimensional photonic crystal consisting of the array of silicon nano-rods, fabricated such photonic crystal on a silicon substrate using reactive ion etching and showed the possibility of its efficient application as a substrate for ELISA detection of human cancer antigens.

[Effects of plastic film mulching and rain harvesting modes on chlorophyll fluorescence characteristics, yield and water use efficiency of dryland maize].

PubMed

Li, Shang-Zhong; Fan, Ting-Lu; Wang, Yong; Zhao, Gang; Wang, Lei; Tang, Xiao-Ming; Dang, Yi; Zhao, Hui

2014-02-01

The differences on chlorophyll fluorescence parameters, yield and water use efficiency of dryland maize were compared among full plastic film mulching on double ridges and planting in catchment furrows (FFDRF), half plastic film mulching on double ridges and planting in catchment furrows (HFDRF), plastic film mulching on ridge and planting in film-side (FS), and flat planting with no plastic film mulching (NM) under field conditions in dry highland of Loess Plateau in 2007-2012. The results showed that fluorescence yield (Fo), the maximum fluorescence yield (Fm), light-adapted fluorescence yield when PS II reaction centers were totally open (F), light-adapted fluorescence yield when PS II reaction centers closed (Fm'), the maximal photochemical efficiency of PS II (Fv/Fm), the actual photochemical efficiency of PS II in the light (Phi PS II), the relative electron transport rate (ETR), photochemical quenching (qP) and non-photochemical quenching (qN) in maize leaves of FFDRF were higher than that of control (NM), and the value of 1-qP was lower than that of control, at 13:00, chlorophyll fluorescence parameters values of FFDRF was significantly higher than control, which were increased by 5.3%, 56.8%, 10.7%, 36.3%, 23.6%, 56.7%, 64.4%, 45.5%, 23.6% and -55.6%, respectively, compared with the control. Yield and water use efficiency of FFDRF were the highest in every year no matter dry year, normal year, humid year and hail disaster year. Average yield and water use efficiency of FFDRF were 12,650 kg x hm(-2) and 40.4 kg x mm(-1) x hm(-2) during 2007-2012, increased by 57.8% and 61.6% compared with the control, respectively, and also significantly higher compared with HFDRF and PS. Therefore, it was concluded that FFDRF had significantly increased the efficiency of light energy conversion and improved the production capacity of dryland maize.

High-efficiency in situ resonant inelastic x-ray scattering (iRIXS) endstation at the Advanced Light Source

DOE PAGES

Qiao, Ruimin; Li, Qinghao; Zhuo, Zengqing; ...

2017-03-17

In this paper, an endstation with two high-efficiency soft x-ray spectrographs was developed at Beamline 8.0.1 of the Advanced Light Source, Lawrence Berkeley National Laboratory. The endstation is capable of performing soft x-ray absorption spectroscopy, emission spectroscopy, and, in particular, resonant inelastic soft x-ray scattering (RIXS). Two slit-less variable line-spacing grating spectrographs are installed at different detection geometries. The endstation covers the photon energy range from 80 to 1500 eV. For studying transition-metal oxides, the large detection energy window allows a simultaneous collection of x-ray emission spectra with energies ranging from the O K-edge to the Ni L-edge without movingmore » any mechanical components. The record-high efficiency enables the recording of comprehensive two-dimensional RIXS maps with good statistics within a short acquisition time. By virtue of the large energy window and high throughput of the spectrographs, partial fluorescence yield and inverse partial fluorescence yield signals could be obtained for all transition metal L-edges including Mn. Finally and moreover, the different geometries of these two spectrographs (parallel and perpendicular to the horizontal polarization of the beamline) provide contrasts in RIXS features with two different momentum transfers.« less

Classic Maximum Entropy Recovery of the Average Joint Distribution of Apparent FRET Efficiency and Fluorescence Photons for Single-molecule Burst Measurements

PubMed Central

DeVore, Matthew S.; Gull, Stephen F.; Johnson, Carey K.

2012-01-01

We describe a method for analysis of single-molecule Förster resonance energy transfer (FRET) burst measurements using classic maximum entropy. Classic maximum entropy determines the Bayesian inference for the joint probability describing the total fluorescence photons and the apparent FRET efficiency. The method was tested with simulated data and then with DNA labeled with fluorescent dyes. The most probable joint distribution can be marginalized to obtain both the overall distribution of fluorescence photons and the apparent FRET efficiency distribution. This method proves to be ideal for determining the distance distribution of FRET-labeled biomolecules, and it successfully predicts the shape of the recovered distributions. PMID:22338694

Heavy Atom Effect of Bromine Significantly Enhances Exciton Utilization of Delayed Fluorescence Luminogens.

PubMed

Gan, Shifeng; Hu, Shimin; Li, Xiang-Long; Zeng, Jiajie; Zhang, Dongdong; Huang, Tianyu; Luo, Wenwen; Zhao, Zujin; Duan, Lian; Su, Shi-Jian; Tang, Ben Zhong

2018-05-23

Raising triplet exciton utilization of pure organic luminescent materials is of significant importance for efficiency advancement of organic light-emitting diodes (OLEDs). Herein, by introducing bromine atom(s) onto a typical molecule (bis(carbazol-9-yl)-4,5-dicyanobenzene) with thermally activated delayed fluorescence, we demonstrate that the heavy atom effect of bromine can increase spin-orbit coupling and promote the reverse intersystem crossing, which endow the molecules with more distinct delayed fluorescence. In consequence, the triplet exciton utilization is improved greatly with the increase of bromine atoms, affording apparently advanced external quantum efficiencies of OLEDs. Utilizing the enhancement effect of bromine atoms on delayed fluorescence should be a simple and promising design concept for efficient organic luminogens with high exciton utilization.

A Leaf-Inspired Luminescent Solar Concentrator for Energy-Efficient Continuous-Flow Photochemistry.

PubMed

Cambié, Dario; Zhao, Fang; Hessel, Volker; Debije, Michael G; Noël, Timothy

2017-01-19

The use of solar light to promote chemical reactions holds significant potential with regard to sustainable energy solutions. While the number of visible light-induced transformations has increased significantly, the use of abundant solar light has been extremely limited. We report a leaf-inspired photomicroreactor that constitutes a merger between luminescent solar concentrators (LSCs) and flow photochemistry to enable green and efficient reactions powered by solar irradiation. This device based on fluorescent dye-doped polydimethylsiloxane collects sunlight, focuses the energy to a narrow wavelength region, and then transports that energy to embedded microchannels where the flowing reactants are converted. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Prediction and design of efficient exciplex emitters for high-efficiency, thermally activated delayed-fluorescence organic light-emitting diodes.

PubMed

Liu, Xiao-Ke; Chen, Zhan; Zheng, Cai-Jun; Liu, Chuan-Lin; Lee, Chun-Sing; Li, Fan; Ou, Xue-Mei; Zhang, Xiao-Hong

2015-04-08

High-efficiency, thermally activated delayed-fluorescence organic light-emitting diodes based on exciplex emitters are demonstrated. The best device, based on a TAPC:DPTPCz emitter, shows a high external quantum efficiency of 15.4%. Strategies for predicting and designing efficient exciplex emitters are also provided. This approach allow prediction and design of efficient exciplex emitters for achieving high-efficiency organic light-emitting diodes, for future use in displays and lighting applications. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Highly efficient red OLEDs using DCJTB as the dopant and delayed fluorescent exciplex as the host

PubMed Central

Zhao, Bo; Zhang, Tianyou; Chu, Bei; Li, Wenlian; Su, Zisheng; Wu, Hairuo; Yan, Xingwu; Jin, Fangming; Gao, Yuan; Liu, Chengyuan

2015-01-01

In this manuscript, we demonstrated a highly efficient DCJTB emission with delayed fluorescent exciplex TCTA:3P-T2T as the host. For the 1.0% DCJTB doped concentration, a maximum luminance, current efficiency, power efficiency and EQE of 22,767 cd m−2, 22.7 cd A−1, 21.5 lm W−1 and 10.15% were achieved, respectively. The device performance is the best compared to either red OLEDs with traditional fluorescent emitter or traditional red phosphor of Ir(piq)3 doped into CBP host. The extraction of so high efficiency can be explained as the efficient triplet excitons up-conversion of TCTA:3P-T2T and the energy transfer from exciplex host singlet state to DCJTB singlet state. PMID:26023882

Relative fluorescent efficiency of sodium salicylate between 90 and 800 eV

NASA Technical Reports Server (NTRS)

Angel, G. C.; Samson, J. A. R.; Wiliams, G.

1986-01-01

The relative fluorescent quantum efficiency of sodium salicylate was measured between 90 and 800 eV (138-15 A) by the use of synchrotron radiation. A general increase in efficiency was observed in this spectral range except for abrupt decreases in efficiency at the carbon and oxygen K-edges. Beyond the oxygen K-edge (532 eV) the efficiency increased linearly with the incident photon energy to the limit of the present observations.

The relative fluorescent efficiency of sodium salicylate between 90 and 800 eV

NASA Technical Reports Server (NTRS)

Angel, G. C.; Samson, J. A. R.; Williams, G.

1986-01-01

The relative fluorescent quantum efficiency of sodium salicylate was measured between 90 and 800 eV (138 -15 A) by the use of synchrotron radiation. A general increase in efficiency was observed in this spectral range except for abrupt decreases in efficiency at the carbon and oxygen K-edges. Beyond the oxygen K-edge (532 eV) the efficiency increased linearly with the incident photon energy to the limit of the present observations.

A system architecture for online data interpretation and reduction in fluorescence microscopy

NASA Astrophysics Data System (ADS)

Röder, Thorsten; Geisbauer, Matthias; Chen, Yang; Knoll, Alois; Uhl, Rainer

2010-01-01

In this paper we present a high-throughput sample screening system that enables real-time data analysis and reduction for live cell analysis using fluorescence microscopy. We propose a novel system architecture capable of analyzing a large amount of samples during the experiment and thus greatly minimizing the post-analysis phase that is the common practice today. By utilizing data reduction algorithms, relevant information of the target cells is extracted from the online collected data stream, and then used to adjust the experiment parameters in real-time, allowing the system to dynamically react on changing sample properties and to control the microscope setup accordingly. The proposed system consists of an integrated DSP-FPGA hybrid solution to ensure the required real-time constraints, to execute efficiently the underlying computer vision algorithms and to close the perception-action loop. We demonstrate our approach by addressing the selective imaging of cells with a particular combination of markers. With this novel closed-loop system the amount of superfluous collected data is minimized, while at the same time the information entropy increases.

Comparison of higher-order multiphoton signal generation and collection at the 1700-nm window based on transmittance measurement of objective lenses.

PubMed

Wen, Wenhui; Wang, Yuxin; Liu, Hongji; Wang, Kai; Qiu, Ping; Wang, Ke

2018-01-01

One benefit of excitation at the 1700-nm window is the more accessible modalities of multiphoton signal generation. It is demonstrated here that the transmittance performance of the objective lens is of vital importance for efficient higher-order multiphoton signal generation and collection excited at the 1700-nm window. Two commonly used objective lenses for multiphoton microscopy (MPM) are characterized and compared, one with regular coating and the other with customized coating for high transmittance at the 1700-nm window. Our results show that, fourth harmonic generation imaging of mouse tail tendon and 5-photon fluorescence of carbon quantum dots using the regular objective lens shows an order of magnitude signal higher than those using the customized objective lens. Besides, the regular objective lens also enables a 3-photon fluorescence imaging depth of >1600 μm in mouse brain in vivo. Our results will provide guidelines for objective lens selection for MPM at the 1700-nm window. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

DNA-length-dependent quenching of fluorescently labeled iron oxide nanoparticles with gold, graphene oxide and MoS2 nanostructures.

PubMed

Balcioglu, Mustafa; Rana, Muhit; Robertson, Neil; Yigit, Mehmet V

2014-08-13

We controlled the fluorescence emission of a fluorescently labeled iron oxide nanoparticle using three different nanomaterials with ultraefficient quenching capabilities. The control over the fluorescence emission was investigated via spacing introduced by the surface-functionalized single-stranded DNA molecules. DNA molecules were conjugated on different templates, either on the surface of the fluorescently labeled iron oxide nanoparticles or gold and nanographene oxide. The efficiency of the quenching was determined and compared with various fluorescently labeled iron oxide nanoparticle and nanoquencher combinations using DNA molecules with three different lengths. We have found that the template for DNA conjugation plays significant role on quenching the fluorescence emission of the fluorescently labeled iron oxide nanoparticles. We have observed that the size of the DNA controls the quenching efficiency when conjugated only on the fluorescently labeled iron oxide nanoparticles by setting a spacer between the surfaces and resulting change in the hydrodynamic size. The quenching efficiency with 12mer, 23mer and 36mer oligonucleotides decreased to 56%, 54% and 53% with gold nanoparticles, 58%, 38% and 32% with nanographene oxide, 46%, 38% and 35% with MoS2, respectively. On the other hand, the presence, not the size, of the DNA molecules on the other surfaces quenched the fluorescence significantly with different degrees. To understand the effect of the mobility of the DNA molecules on the nanoparticle surface, DNA molecules were attached to the surface with two different approaches. Covalently immobilized oligonucleotides decreased the quenching efficiency of nanographene oxide and gold nanoparticles to ∼22% and ∼21%, respectively, whereas noncovalently adsorbed oligonucleotides decreased it to ∼25% and ∼55%, respectively. As a result, we have found that each nanoquencher has a powerful quenching capability against a fluorescent nanoparticle, which can be tuned with surface functionalized DNA molecules.

Thermally Activated Delayed Fluorescence in Polymers: A New Route toward Highly Efficient Solution Processable OLEDs.

PubMed

Nikolaenko, Andrey E; Cass, Michael; Bourcet, Florence; Mohamad, David; Roberts, Matthew

2015-11-25

Efficient intermonomer thermally activated delayed fluorescence is demonstrated for the first time, opening a new route to achieving high-efficiency solution processable polymer light-emitting device materials. External quantum efficiency (EQE) of up to 10% is achieved in a simple fully solution-processed device structure, and routes for further EQE improvement identified. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

An Analysis of Sources of Technological Change in Efficiency Improvement of Fluorescent Lamp Systems

NASA Astrophysics Data System (ADS)

Imanaka, Takeo

In Japan, energy efficient fluorescent lamp systems which use “rare-earth phosphors” and “electronic ballasts” have shown rapid diffusion since 1990s. This report investigated sources of technological change in the efficiency improvement of fluorescent lamp systems: (i) Fluorescent lamp and luminaires have been under steady technological development for getting more energy efficient lighting and the concepts to achieve high efficiency had been found in such activities; however, it took long time until they realized and become widely used; (ii) Electronic ballasts and rare-earth phosphors add fluorescent lamp systems not only energy efficiency but also various values such as compactness, lightweight, higher output, and better color rendering properties, which have also been expected and have induced research and development (R&D) (iii) Affordable electronic ballasts are realized by the new technology “power MOSFET” which is based on IC technologies and has been developed for large markets of information and communication technologies and mobile devices; and (iv) Rare-earth phosphors became available after rare-earth industries developed for the purpose of supplying rare-earth phosphors for color television. In terms of sources of technological change, (i) corresponds to “R&D” aiming at the particular purpose i.e. energy efficiency in this case, on the other hand, (ii), (iii), and (iv) correspond to “spillovers” from activities aiming at other purposes. This case exhibits an actual example in which “spillovers” were the critical sources of technological change in energy technology.

FRET Sensor for Erythrosine Dye Based on Organic Nanoparticles: Application to Analysis of Food Stuff.

PubMed

Mahajan, Prasad G; Bhopate, Dhanaji P; Kolekar, Govind B; Patil, Shivajirao R

2016-07-01

An aqueous suspension of fluorescent nanoparticles (PHNNPs) of naphthol based fluorescent organic compound 1-[(Z)-(2-phenylhydrazinylidene) methyl] naphthalene -2-ol (PHN) were prepared using reprecipitation method shows bathochromically shifted aggregation induced enhanced emission (AIEE) in the spectral region where erythrosine (ETS) food dye absorbs strongly. The average size of 72.6 nm of aqueous suspension of PHNNPs obtained by Dynamic light scattering results shows a narrow particle size distribution. The negative zeta potential of nano probe (-22.6 mV) responsible to adsorb oppositely charged analyte on its surface and further permit to bind nano probe and analyte within the close distance proximity required for efficient fluorescence resonance energy transfer (FRET) to take place from donor (PHNNPs) to acceptor (ETS). Systematic FRET experiments performed by measuring fluorescence quenching of PHNNPs with successive addition of ETS solution exploited the use of the PHNNPs as a novel nano probe for the detection of ETS in aqueous solution with extremely lower limit of detection equal to 3.6 nM (3.1 ng/mL). The estimation of photo kinetic and thermodynamic parameters such as quenching rate constant, enthalpy change (∆H), Gibbs free energy change (∆G) and entropy change (∆S) was obtained by the quenching results obtained at different constant temperatures which were found to fit the well-known Stern-Volmer relation. The mechanism of binding and fluorescence quenching of PHNNPs by ETS food dye is proposed on the basis of results obtained in photophysical studies, thermodynamic parameter, energy transfer efficiency, critical energy transfer distance (R0) and distance of approach between donor-acceptor molecules (r). The proposed FRET method based on fluorescence quenching of PHNNPs was successfully applied to develop an analytical method for estimation of ETS from food stuffs without interference of other complex ingredients. Graphical Abstract A fluorescent organic nanoprobe developed for the detection of erythrosine (ETS) food dye in aqueous medium based on fluorescence resonance energy transfer (FRET). The FRET process between donor (nanoparticles) and acceptor (ETS dye) arises due to oppositely charge attraction through hydrophobic interactions. The proposed method was successfully applied to quantitative determination of ETS dye in food stuff sample collected from local market.

Metal plasmon-coupled fluorescence imaging and label free coenzyme detection in cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Jian, E-mail: jian@cfs.bioment.umaryland.edu; Fu, Yi; Li, Ge

2012-08-31

Highlights: Black-Right-Pointing-Pointer Metal nanoparticle for fluorescence cell imaging. Black-Right-Pointing-Pointer Non-invasive emission detection of coenzyme in cell on time-resolved confocal microscope. Black-Right-Pointing-Pointer Near-field interaction of flavin adenine dinucleotide with silver substrate. Black-Right-Pointing-Pointer Isolation of emissions by coenzymes from cellular autofluorescence on fluorescence cell imaging. -- Abstract: Flavin adenine dinucleotide (FAD) is a key metabolite in cellular energy conversion. Flavin can also bind with some enzymes in the metabolic pathway and the binding sites may be changed due to the disease progression. Thus, there is interest on studying its expression level, distribution, and redox state within the cells. FAD is naturally fluorescent,more » but it has a modest extinction coefficient and quantum yield. Hence the intrinsic emission from FAD is generally too weak to be isolated distinctly from the cellular backgrounds in fluorescence cell imaging. In this article, the metal nanostructures on the glass coverslips were used as substrates to measure FAD in cells. Particulate silver films were fabricated with an optical resonance near the absorption and the emission wavelengths of FAD which can lead to efficient coupling interactions. As a result, the emission intensity and quantum yield by FAD were greatly increased and the lifetime was dramatically shortened resulting in less interference from the longer lived cellular background. This feature may overcome the technical limits that hinder the direct observation of intrinsically fluorescent coenzymes in the cells by fluorescence microscopy. Fluorescence cell imaging on the metallic particle substrates may provide a non-invasive strategy for collecting the information of coenzymes in cells.« less

JOVE Program

NASA Technical Reports Server (NTRS)

Hardy, John T.

1997-01-01

Coastal reef degradation and widespread bleaching of corals, i.e. loss of pigments and/or symbiotic zooxanthellae, is increasing globally. Remote sensing has great potential for assessing the extent of reef change. However, measuring reef change from remote platforms (boats, aircraft or satellites), requires ground-truth spectral algorithms characteristic of healthy and degraded reef populations. Our laboratory experiments demonstrated, for the first time, that healthy coral colonies emit characteristic fluorescence spectra in response to excitation with short wavelength (blue or ultraviolet) light. Furthermore, following stress, e.g. increased temperature, corals lose symbiotic algae and their health is compromised. We measured changes in coral fluorescence spectra along with pigment loss in response to temperature-induced stress. Following laboratory demonstration, we tested this approach in the field. The NASA P3 aircraft with the airborne oceanographic LIDAR system was used to map transects of coral reef in Hawaii. Ground truth samples of coral; were collected and analyzed for pigment density (pigments/surface area). Fluorescence spectra along transects showed consistent chlorophyll fluorescence peaks at 685 nm from the chlorophyll of the zooxanthellae and peaks between 400 and 570 nm believed to emanate from the coral tissue. Our results suggest that remote sensing of laser-induced fluorescence represents an efficient and effective approach to monitoring the health of coral reefs. Lists of papers, conferences, proposals awarded and a summary of student involvement is included in the appendices.

Nanostructured Surfaces and Detection Instrumentation for Photonic Crystal Enhanced Fluorescence

PubMed Central

Chaudhery, Vikram; George, Sherine; Lu, Meng; Pokhriyal, Anusha; Cunningham, Brian T.

2013-01-01

Photonic crystal (PC) surfaces have been demonstrated as a compelling platform for improving the sensitivity of surface-based fluorescent assays used in disease diagnostics and life science research. PCs can be engineered to support optical resonances at specific wavelengths at which strong electromagnetic fields are utilized to enhance the intensity of surface-bound fluorophore excitation. Meanwhile, the leaky resonant modes of PCs can be used to direct emitted photons within a narrow range of angles for more efficient collection by a fluorescence detection system. The multiplicative effects of enhanced excitation combined with enhanced photon extraction combine to provide improved signal-to-noise ratios for detection of fluorescent emitters, which in turn can be used to reduce the limits of detection of low concentration analytes, such as disease biomarker proteins. Fabrication of PCs using inexpensive manufacturing methods and materials that include replica molding on plastic, nano-imprint lithography on quartz substrates result in devices that are practical for single-use disposable applications. In this review, we will describe the motivation for implementing high-sensitivity fluorescence detection in the context of molecular diagnosis and gene expression analysis though the use of PC surfaces. Recent efforts to improve the design and fabrication of PCs and their associated detection instrumentation are summarized, including the use of PCs coupled with Fabry-Perot cavities and external cavity lasers. PMID:23624689

Sun-Induced Chlorophyll Fluorescence, Photosynthesis, and Light Use Efficiency of a Soybean Field from Seasonally Continuous Measurements

DOE PAGES

Miao, Guofang; Guan, Kaiyu; Yang, Xi; ...

2018-01-29

Recent development of sun-induced chlorophyll fluorescence (SIF) technology is stimulating studies to remotely approximate canopy photosynthesis (measured as gross primary production, GPP). While multiple applications have advanced the empirical relationship between GPP and SIF, mechanistic understanding of this relationship is still limited. GPP:SIF relationship, using the standard light use efficiency framework, is determined by absorbed photosynthetically active radiation (APAR) and the relationship between photosynthetic light use efficiency (LUE) and fluorescence yield (SIF y). While previous studies have found that APAR is the dominant factor of the GPP:SIF relationship, the LUE:SIF y relationship remains unclear. For a better understanding of themore » LUE:SIF y relationship, in this paper we deployed a ground-based system (FluoSpec2), with an eddy-covariance flux tower at a soybean field in the Midwestern U.S. during the 2016 growing season to collect SIF and GPP data simultaneously. With the measurements categorized by plant growth stages, light conditions, and time scales, we confirmed that a strong positive GPP:SIF relationship was dominated by an even stronger linear SIF:APAR relationship. By normalizing both GPP and SIF by APAR, we found that under sunny conditions our soybean field exhibited a clear positive SIF y:APAR relationship and a weak negative LUE:SIF y relationship, opposite to the positive LUE:SIF y relationship reported previously in other ecosystems. Our study provides a first continuous SIF record over multiple growth stages for agricultural systems and reveals a distinctive pattern related to the LUE:SIF y relationship compared with previous work. Finally, the observed positive relationship of SIF y:APAR at the soybean site provides new insights of the previous understanding on the SIF's physiological implications.« less

Sun-Induced Chlorophyll Fluorescence, Photosynthesis, and Light Use Efficiency of a Soybean Field from Seasonally Continuous Measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miao, Guofang; Guan, Kaiyu; Yang, Xi

Recent development of sun-induced chlorophyll fluorescence (SIF) technology is stimulating studies to remotely approximate canopy photosynthesis (measured as gross primary production, GPP). While multiple applications have advanced the empirical relationship between GPP and SIF, mechanistic understanding of this relationship is still limited. GPP:SIF relationship, using the standard light use efficiency framework, is determined by absorbed photosynthetically active radiation (APAR) and the relationship between photosynthetic light use efficiency (LUE) and fluorescence yield (SIF y). While previous studies have found that APAR is the dominant factor of the GPP:SIF relationship, the LUE:SIF y relationship remains unclear. For a better understanding of themore » LUE:SIF y relationship, in this paper we deployed a ground-based system (FluoSpec2), with an eddy-covariance flux tower at a soybean field in the Midwestern U.S. during the 2016 growing season to collect SIF and GPP data simultaneously. With the measurements categorized by plant growth stages, light conditions, and time scales, we confirmed that a strong positive GPP:SIF relationship was dominated by an even stronger linear SIF:APAR relationship. By normalizing both GPP and SIF by APAR, we found that under sunny conditions our soybean field exhibited a clear positive SIF y:APAR relationship and a weak negative LUE:SIF y relationship, opposite to the positive LUE:SIF y relationship reported previously in other ecosystems. Our study provides a first continuous SIF record over multiple growth stages for agricultural systems and reveals a distinctive pattern related to the LUE:SIF y relationship compared with previous work. Finally, the observed positive relationship of SIF y:APAR at the soybean site provides new insights of the previous understanding on the SIF's physiological implications.« less

Drought is Coming: Monitoring Vegetation Response to Water Scarcity through Variable Chlorophyll a Fluorescence

NASA Astrophysics Data System (ADS)

Guadagno, C. R.; Beverly, D.; Pleban, J. R.; Speckman, H. N.; Ewers, B. E.; Weinig, C.

2017-12-01

Aridity is one of the most pronounced environmental limits to plant survival, and understanding how plants respond to drought and recovery is crucial for predicting impacts on managed and natural ecosystems. Changes in soil moisture conditions induce a suite of physiological responses from the cell to ecosystem scale, complicating the assessment of drought effects. Characterizing early indicators of water scarcity across species can inform biophysical models with improved understanding of plant hydraulics. While indexes exist for drought monitoring across scales, many are unable to identify imminent vegetative drought. We explore a method of early diagnosis using leaf-level and kinetic imaging measures of variable chlorophyll a fluorescence. This is a fast and reliable tool capturing leaf physiological changes in advance of changes in NDVI or passive solar induced fluorescence. Both image and leaf level Pulse Amplitude Method (PAM) measurements illustrate the utility of variable chlorophyll a fluorescence for monitoring vegetative drought. Variable fluorescence was monitored across populations of crops, desert shrubs, montane conifers and riparian deciduous trees under variable water regimes. We found a strong correlation (R = 0.85) between the maximum efficiency of photosystem II measured using variable fluorescence (Fv'Fm') and leaf level electrolyte leakage, a proximal cause of drought stress induced by cellular damage in leaves. This association was confirmed in two gymnosperm species (Picea engelmannii and Pinus contorta) and for diverse varieties of the crop species Brassica rapa. The use of chlorophyll a fluorescence per image also allowed for early detection of drought in aspen (Populus tremuloides). These results provide evidence that variable chlorophyll fluorescence decreases between 25% and 70% in mild and severely droughted twigs with respect to ones collected from trees in wet soil conditions. While current systems for monitoring variable fluorescence are limited in scale, chlorophyll fluorescence comprises an indicator of drought stress across multiple spatial scales, from leaf to ecosystem level.

Near-isotropic 3D optical nanoscopy with photon-limited chromophores

PubMed Central

Tang, Jianyong; Akerboom, Jasper; Vaziri, Alipasha; Looger, Loren L.; Shank, Charles V.

2010-01-01

Imaging approaches based on single molecule localization break the diffraction barrier of conventional fluorescence microscopy, allowing for bioimaging with nanometer resolution. It remains a challenge, however, to precisely localize photon-limited single molecules in 3D. We have developed a new localization-based imaging technique achieving almost isotropic subdiffraction resolution in 3D. A tilted mirror is used to generate a side view in addition to the front view of activated single emitters, allowing their 3D localization to be precisely determined for superresolution imaging. Because both front and side views are in focus, this method is able to efficiently collect emitted photons. The technique is simple to implement on a commercial fluorescence microscope, and especially suitable for biological samples with photon-limited chromophores such as endogenously expressed photoactivatable fluorescent proteins. Moreover, this method is relatively resistant to optical aberration, as it requires only centroid determination for localization analysis. Here we demonstrate the application of this method to 3D imaging of bacterial protein distribution and neuron dendritic morphology with subdiffraction resolution. PMID:20472826

Metal–Dielectric Waveguides for High Efficiency Fluorescence Imaging

PubMed Central

Zhu, Liangfu; Zhang, Douguo; Wang, Ruxue; Wang, Pei; Ming, Hai; Badugu, Ramachandram; Du, Luping; Yuan, Xiaocong; Lakowicz, Joseph R.

2015-01-01

We demonstrate that Metal–Dielectric Waveguide structures (MDWs) with high efficiency of fluorescence coupling can be suitable as substrates for fluorescence imaging. This hybrid MDWs consists of a continuous metal film and a dielectric top layer. The optical modes sustaining inside this structure can be excited with a high numerical aperture (N.A) objective, and then focused into a virtual optical probe with high intensity, leading to efficient excitation of fluorophores deposited on top of the MDWs. The emitted fluorophores couple with the optical modes thus enabling the directional emission, which is verified by the back focal plane (BFP) imaging. These unique properties of MDWs have been adopted in a scanning laser confocal optical microscopy, and show the merit of high efficiency fluorescence imaging. MDWs can be easily fabricated by vapor deposition and/or spin coating, the silica surface of the MDWs is suitable for biomolecule tethering, and will offer new opportunities for cell biology and biophysics research. PMID:26525494

Scanning fluorescent microthermal imaging apparatus and method

DOEpatents

Barton, Daniel L.; Tangyunyong, Paiboon

1998-01-01

A scanning fluorescent microthermal imaging (FMI) apparatus and method is disclosed, useful for integrated circuit (IC) failure analysis, that uses a scanned and focused beam from a laser to excite a thin fluorescent film disposed over the surface of the IC. By collecting fluorescent radiation from the film, and performing point-by-point data collection with a single-point photodetector, a thermal map of the IC is formed to measure any localized heating associated with defects in the IC.

Triplet-triplet annihilation in highly efficient fluorescent organic light-emitting diodes: current state and future outlook.

PubMed

Kondakov, Denis Y

2015-06-28

Studies of delayed electroluminescence in highly efficient fluorescent organic light-emitting diodes (OLEDs) of many dissimilar architectures indicate that the triplet-triplet annihilation (TTA) significantly increases yield of excited singlet states-emitting molecules in this type of device thereby contributes substantially to their efficiency. Towards the end of the 2000s, the essential role of TTA in realizing highly efficient fluorescent devices was widely recognized. Analysis of a diverse set of fluorescent OLEDs shows that high efficiencies are often cor-related to TTA extents. It is therefore likely that it is the long-term empirical optimization of OLED efficiencies that has resulted in fortuitous emergence of TTA as a large and ubiquitous contributor to efficiency. TTA contributions as high as 20-30% are common in the state-of-the-art OLEDs, and even become dominant in special cases, where TTA is shown to substantially exceed the spin-statistical limit. The fundamental features of OLED efficiency enhancement via TTA-molecular structure-dependent contributions, current density-dependent intensities in practical devices and frequently observed antagonistic relationships between TTA extent and OLED lifetime-came to be understood over the course of the next few years. More recently, however, there was much less reported progress with respect to all-important quantitative details of the TTA mechanism. It should be emphasized that, to this day and despite the decades of work on improving blue phosphorescent OLEDs as well as the recent advent of thermally activated delayed fluorescence OLEDs, the majority of practical blue OLEDs still rely on TTA. Considering such practical importance of fluorescent blue OLEDs, the design of blue OLED-compatible materials capable of substantially exceeding the spin-statistical limit in TTA, elimination of the antagonistic relationship between TTA-related efficiency gains and lifetime losses, and designing devices with an extended range of current densities producing near-maximum TTA electroluminescence are the areas where future improvements would be most beneficial. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Diffuse reflectance of the ocean - The theory of its augmentation by chlorophyll a fluorescence at 685 nm

NASA Technical Reports Server (NTRS)

Gordon, H. R.

1979-01-01

The radiative transfer equation is modified to include the effect of fluorescent substances and solved in the quasi-single scattering approximation for a homogeneous ocean containing fluorescent particles with wavelength independent quantum efficiency and a Gaussian shaped emission line. The results are applied to the in vivo fluorescence of chlorophyll a (in phytoplankton) in the ocean to determine if the observed quantum efficiencies are large enough to explain the enhancement of the ocean's diffuse reflectance near 685 nm in chlorophyll rich waters without resorting to anomalous dispersion. The computations indicate that the required efficiencies are sufficiently low to account completely for the enhanced reflectance. The validity of the theory is further demonstrated by deriving values for the upwelling irradiance attenuation coefficient at 685 nm which are in close agreement with the observations.

Detection and Quantification of 8-Hydroxy-2′-Deoxyguanosine in Alzheimer’s Transgenic Mouse Urine using Capillary Electrophoresis

PubMed Central

Zhang, Cheng; Nestorova, Gergana; Rissman, Robert A.; Feng, June

2013-01-01

8-Hydroxy-2′-deoxyguanosine (8-OHdG) is one of the major forms of oxidative deoxyribonucleic acid (DNA) damage, and is commonly analyzed as an excellent marker of DNA lesions. The purpose of this study was to develop a sensitive method to accurately and rapidly quantify the 8-OHdG by using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). The method involved the use of specific antibody to detect DNA lesions (8-OHdG) and consecutive fluorescence labeling. Next, the urine sample with 8-OHdG fluorescently labeled along with other constituents was resolved by capillary electrophoretic system and the lesion of interest was detected using fluorescence detector. The limit of detection was 0.18 fmol, which is sufficient sensitivity for detection and quantification of 8-OHdG in untreated urine samples. The relative standard deviation (RSD) was found to be 11.32 % for migration time, and 5.52 % for peak area. To demonstrate the utility of this method, the urinary concentration of 8-OHdG in an Alzheimer’s transgenic mouse model was determined. Collectively, our results indicate that this methodology offers great advantages such as high separation efficiency, good selectivity, low limit of detection (LOD), simplicity and low cost of analysis. PMID:23712533

Compact 3D printed module for fluorescence and label-free imaging using evanescent excitation

NASA Astrophysics Data System (ADS)

Pandey, Vikas; Gupta, Shalini; Elangovan, Ravikrishnan

2018-01-01

Total internal reflection fluorescence (TIRF) microscopy is widely used for selective excitation and high-resolution imaging of fluorophores, and more recently label-free nanosized objects, with high vertical confinement near a liquid-solid interface. Traditionally, high numerical aperture objectives (>1.4) are used to simultaneously generate evanescent waves and collect fluorescence emission signals which limits their use to small area imaging (<0.1 mm2). Objective-based TIRFs are also expensive as they require dichroic mirrors and efficient notch filters to prevent specular reflection within the objective lenses. We have developed a compact 3D module called cTIRF that can generate evanescent waves in microscope glass slides via a planar waveguide illumination. The module can be attached as a fixture to any existing optical microscope, converting it into a TIRF and enabling high signal-to-noise ratio (SNR) fluorescence imaging using any magnification objective. As the incidence optics is perpendicular to the detector, label-free evanescent scattering-based imaging of submicron objects can also be performed without using emission filters. SNR is significantly enhanced in this case as compared to cTIRF alone, as seen through our model experiments performed on latex beads and mammalian cells. Extreme flexibility and the low cost of our approach makes it scalable for limited resource settings.

Radioisotope Detection Device and Methods of Radioisotope Collection

DOEpatents

Tranter, Troy J [Idaho Falls, ID; Oertel, Christopher P [Idaho Falls, ID; Giles, John R [Pocatello, ID; Mann, Nicholas R [Rigby, ID; McIlwain, Michael E [Idaho Falls, ID

2011-04-12

A device for collection of radionuclides includes a mixture of a polymer, a fluorescent organic scintillator and a chemical extractant. A radionuclide detector system includes a collection device comprising a mixture of a polymer, a fluorescent agent and a selective ligand. The system includes at least one photomultiplier tube (PMT). A method of detecting radionuclides includes providing a collector device comprising a mixture comprising a polymer, a fluorescent organic scintillator and a chemical extractant. An aqueous environment is exposed to the device and radionuclides are collected from the environment. Radionuclides can be concentrated within the device.

Fluorescent-Magnetic-Biotargeting Multifunctional Nanobioprobes for Detecting and Isolating Multiple Types of Tumor Cells

PubMed Central

Song, Er-Qun; Hu, Jun; Wen, Cong-Ying; Tian, Zhi-Quan; Yu, Xu; Zhang, Zhi-Ling; Shi, Yun-Bo; Pang, Dai-Wen

2011-01-01

Fluorescent-magnetic-biotargeting multifunctional nanobioprobes (FMBMNs) have attracted great attention in recent years due to their increasing, important applications in biomedical research, clinical diagnosis, and biomedicine. We have previously developed such nanobioprobes for the detection and isolation of a single kind of tumor cells. Detection and isolation of multiple tumor markers or tumor cells from complex samples sensitively and with high efficiency is critical for the early diagnosis of tumors, especially malignant tumors or cancers, which will improve clinical diagnosis outcomes and help to select effective treatment approaches. Here, we expanded the application of the monoclonal antibody (mAb)-coupled FMBMNs for multiplexed assays. Multiple types of cancer cells, such as leukemia cells and prostate cancer cells, were detected and collected from mixed samples within 25 minutes by using a magnet and an ordinary fluorescence microscope. The capture efficiencies of mAb-coupled FMBMNs for the above mentioned two types of cells were 96% and 97% respectively. Furthermore, by using the mAb-coupled FMBMNs, specific and sensitive detection and rapid separation of a small number of spiked leukemia cells and prostate cancer cells in a large population of cultured normal cells (about 0.01% were tumor cells) were achieved simply and inexpensively without any sample pretreatment before cell analysis. Therefore, mAb-coupled multicolour FMBMNs may be used for very sensitive detection and rapid isolation of multiple cancer cells in biomedical research and medical diagnostics. PMID:21250650

Efficient fluorescence energy transfer system between CdTe-doped silica nanoparticles and gold nanoparticles for turn-on fluorescence detection of melamine.

PubMed

Gao, Feng; Ye, Qingqing; Cui, Peng; Zhang, Lu

2012-05-09

We here report an efficient and enhanced fluorescence energy transfer system between confined quantum dots (QDs) by entrapping CdTe into the mesoporous silica shell (CdTe@SiO₂) as donors and gold nanoparticles (AuNPs) as acceptors. At pH 6.50, the CdTe@SiO₂-AuNPs assemblies coalesce to form larger clusters due to charge neutralization, leading to the fluorescence quenching of CdTe@SiO₂ as a result of energy transfer. As compared with the energy transfer system between unconfined CdTe and AuNPs, the maximum fluorescence quenching efficiency of the proposed system is improved by about 27.0%, and the quenching constant, K(sv), is increased by about 2.4-fold. The enhanced quenching effect largely turns off the fluorescence of CdTe@SiO₂ and provides an optimal "off-state" for sensitive "turn-on" assay. In the present study, upon addition of melamine, the weak fluorescence system of CdTe@SiO₂-AuNPs is enhanced due to the strong interactions between the amino group of melamine and the gold nanoparticles via covalent bond, leading to the release of AuNPs from the surfaces of CdTe@SiO₂; thus, its fluorescence is restored. A "turn-on" fluorimetric method for the detection of melamine is proposed based on the restored fluorescence of the system. Under the optimal conditions, the fluorescence enhanced efficiency shows a linear function against the melamine concentrations ranging from 7.5 × 10⁻⁹ to 3.5 × 10⁻⁷ M (i.e., 1.0-44 ppb). The analytical sensitivity is improved by about 50%, and the detection limit is decreased by 5.0-fold, as compared with the analytical results using the CdTe-AuNPs system. Moreover, the proposed method was successfully applied to the determination of melamine in real samples with excellent recoveries in the range from 97.4 to 104.1%. Such a fluorescence energy transfer system between confined QDs and AuNPs may pave a new way for designing chemo/biosensing.

Fluorescence Quantum Yield Measurements of Fluorescent Proteins: A Laboratory Experiment for a Biochemistry or Molecular Biophysics Laboratory Course

ERIC Educational Resources Information Center

Wall, Kathryn P.; Dillon, Rebecca; Knowles, Michelle K.

2015-01-01

Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts…

Scanning fluorescent microthermal imaging apparatus and method

DOEpatents

Barton, D.L.; Tangyunyong, P.

1998-01-06

A scanning fluorescent microthermal imaging (FMI) apparatus and method is disclosed, useful for integrated circuit (IC) failure analysis, that uses a scanned and focused beam from a laser to excite a thin fluorescent film disposed over the surface of the IC. By collecting fluorescent radiation from the film, and performing point-by-point data collection with a single-point photodetector, a thermal map of the IC is formed to measure any localized heating associated with defects in the IC. 1 fig.

Plasma-Based Mixing Actuation in Airflow, Quantitated by Probe Breakdown Fluorescence

NASA Astrophysics Data System (ADS)

Leonov, Sergey; Firsov, Alexander; Shurupov, Michail; Yarantsev, Dmitry; Ohio State University Team; JIHT RAS Team

2013-09-01

Effective mixing of fuel and oxidizer in air-breathing engine at compressible conditions is an essential problem of high-speed combustion due to short residence time of gas mixture in the combustor of limited length. The effect of the mixing actuation by plasma is observed because of the gasdynamic instability arisen after the long filamentary discharge of submicrosecond duration generated along the contact zone of two co-flown gases. The work is focused on detail consideration of the mechanism of gas instability, promoted by plasma, on effect of the discharge specific localization, and on diagnostics development for qualitative and quantitative estimation of the mixing efficiency. The dynamics of relative concentration of gas components is examined quantitatively by means of Probe Discharge Breakdown Fluorescence (PBF). In this method an optical emission spectra of weak filamentary high-voltage nanosecond probe discharge are collected from local zone of interest in airflow. The first measurements of the mixing efficiency in vicinity of wall-injected secondary gas are presented. It is shown that the method of PBF could deliver experimental data on state of the two-component medium with <1 mcs and <5 mm of time and spatial resolution, correspondingly. Funded by AFOSR under Dr Chiping Li supervision

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qiao, Ruimin; Li, Qinghao; Zhuo, Zengqing

In this paper, an endstation with two high-efficiency soft x-ray spectrographs was developed at Beamline 8.0.1 of the Advanced Light Source, Lawrence Berkeley National Laboratory. The endstation is capable of performing soft x-ray absorption spectroscopy, emission spectroscopy, and, in particular, resonant inelastic soft x-ray scattering (RIXS). Two slit-less variable line-spacing grating spectrographs are installed at different detection geometries. The endstation covers the photon energy range from 80 to 1500 eV. For studying transition-metal oxides, the large detection energy window allows a simultaneous collection of x-ray emission spectra with energies ranging from the O K-edge to the Ni L-edge without movingmore » any mechanical components. The record-high efficiency enables the recording of comprehensive two-dimensional RIXS maps with good statistics within a short acquisition time. By virtue of the large energy window and high throughput of the spectrographs, partial fluorescence yield and inverse partial fluorescence yield signals could be obtained for all transition metal L-edges including Mn. Finally and moreover, the different geometries of these two spectrographs (parallel and perpendicular to the horizontal polarization of the beamline) provide contrasts in RIXS features with two different momentum transfers.« less

A fluorescent probe for ecstasy.

PubMed

Masseroni, D; Biavardi, E; Genovese, D; Rampazzo, E; Prodi, L; Dalcanale, E

2015-08-18

A nanostructure formed by the insertion in silica nanoparticles of a pyrene-derivatized cavitand, which is able to specifically recognize ecstasy in water, is presented. The absence of effects from interferents and an efficient electron transfer process occurring after complexation of ecstasy, makes this system an efficient fluorescent probe for this popular drug.

Hybrid reflecting objectives for functional multiphoton microscopy in turbid media

PubMed Central

Vučinić, Dejan; Bartol, Thomas M.; Sejnowski, Terrence J.

2010-01-01

Most multiphoton imaging of biological specimens is performed using microscope objectives optimized for high image quality under wide-field illumination. We present a class of objectives designed de novo without regard for these traditional constraints, driven exclusively by the needs of fast multiphoton imaging in turbid media: the delivery of femtosecond pulses without dispersion and the efficient collection of fluorescence. We model the performance of one such design optimized for a typical brain-imaging setup and show that it can greatly outperform objectives commonly used for this task. PMID:16880851

On the use of nonfluorescent dye labeled ligands in FRET-based receptor binding studies.

PubMed

Tahtaoui, Chouaib; Guillier, Fabrice; Klotz, Philippe; Galzi, Jean-Luc; Hibert, Marcel; Ilien, Brigitte

2005-12-01

The efficiency of fluorescence resonance energy transfer (FRET) is dependent upon donor-acceptor proximity and spectral overlap, whether the acceptor partner is fluorescent or not. We report here on the design, synthesis, and characterization of two novel pirenzepine derivatives that were coupled to patent blue VF and pinacyanol dyes. These nonfluorescent compounds, when added to cells stably expressing enhanced green fluorescent protein (EGFP)-fused muscarinic M1 receptors, promote EGFP fluorescence extinction in a time-, concentration-, and atropine-dependent manner. They display nanomolar affinity for the muscarinic receptor, determined using either FRET or classical radioligand binding conditions. We provide evidence that these compounds behave as potent acceptors of energy from excited EGFP with quenching efficiencies comparable to those of analogous fluorescent bodipy or rhodamine red pirenzepine derivatives. The advantages they offer over fluorescent ligands are illustrated and discussed in terms of reliability, sensitivity, and wider applicability of FRET-based receptor binding assays.

A Novel Sensitive Luminescence Probe Microspheres for Rapid and Efficient Detection of τ-Fluvalinate in Taihu Lake

PubMed Central

Wang, Jixiang; Wang, Yunyun; Qiu, Hao; Sun, Lin; Dai, Xiaohui; Pan, Jianming; Yan, Yongsheng

2017-01-01

Fluorescent molecularly imprinted polymers have shown great promise in biological or chemical separations and detection, due to their high stability, selectivity and sensitivity. In this work, fluorescent molecularly imprinted microsphere was synthesized via precipitation polymerization, which could separate efficiently and rapidly detect τ-fluvalinate (a toxic insecticide) in water samples, was reported. The fluorescent imprinted sensor showed excellent stability, outstanding selectivity and the limit of detection low to 12.14 nM, good regeneration ability which still kept good sensitivity after 8 cycling experiments and fluorescence quenching mechanism was illustrated in details. In addition, the fluorescent sensor was further used to detect τ-fluvalinate in real samples from Taihu Lake. Despite the relatively complex components of the environment water, the fluorescent imprinted microspheres sitll showed good recovery, clearly demonstrating the potental value of this smart sensor nanomaterial in environment monitoring. PMID:28485402

Method and apparatus for enhanced evanescent fluorescence and color filtering using a high refractive index thin film coating

DOEpatents

Kao, Hung Pin; Schoeniger, Joseph; Yang, Nancy

2001-01-01

A technique for increasing the excitation and collection of evanescent fluorescence radiation emanating from a fiber optic sensor having a high refractive index (n.sub.r), dielectric thin film coating has been disclosed and described. The invention comprises a clad optical fiber core whose cladding is removed on a distal end, the distal end coated with a thin, non-porous, titanium dioxide sol-gel coating. It has been shown that such a fiber will exhibit increased fluorescence coupling due in part by 1) increasing the intensity of the evanescent field at the fiber core surface by a constructive interference effect on the propagating light, and 2) increasing the depth of penetration of the field in the sample. The interference effect created by the thin film imposes a wavelength dependence on the collection of the fluorescence and also suggests a novel application of thin films for color filtering as well as increasing collected fluorescence in fiber sensors. Collected fluorescence radiation increased by up to 6-fold over that of a bare fused silica fiber having a numerical aperture (N.A.) of O.6.

Superior optical nonlinearity of an exceptional fluorescent stilbene dye

DOE Office of Scientific and Technical Information (OSTI.GOV)

He, Tingchao; Division of Physics and Applied Physics, Centre for Disruptive Photonic Technologies; Sreejith, Sivaramapanicker

2015-03-16

Strong multiphoton absorption and harmonic generation in organic fluorescent chromophores are, respectively, significant in many fields of research. However, most of fluorescent chromophores fall short of the full potential due to the absence of the combination of such different nonlinear upconversion behaviors. Here, we demonstrate that an exceptional fluorescent stilbene dye could exhibit efficient two- and three-photon absorption under the excitation of femtosecond pulses in solution phase. Benefiting from its biocompatibility and strong excited state absorption behavior, in vitro two-photon bioimaging and superior optical limiting have been exploited, respectively. Simultaneously, the chromophore could generate efficient three-photon excited fluorescence and third-harmonicmore » generation (THG) when dispersed into PMMA film, circumventing the limitations of classical fluorescent chromophores. Such chromophore may find application in the production of coherent light sources of higher photon energy. Moreover, the combination of three-photon excited fluorescence and THG can be used in tandem to provide complementary information in biomedical studies.« less

Molecularly imprinted fluorescent hollow nanoparticles as sensors for rapid and efficient detection λ-cyhalothrin in environmental water.

PubMed

Wang, Jixiang; Qiu, Hao; Shen, Hongqiang; Pan, Jianming; Dai, Xiaohui; Yan, Yongsheng; Pan, Guoqing; Sellergren, Börje

2016-11-15

Molecularly imprinted fluorescent polymers have shown great promise in biological or chemical separations and detections, due to their high stability, selectivity and sensitivity. In this work, molecularly imprinted fluorescent hollow nanoparticles, which could rapidly and efficiently detect λ-cyhalothrin (a toxic insecticide) in water samples, was reported. The molecularly imprinted fluorescent sensor showed excellent sensitivity (the limit of detection low to 10.26nM), rapid detection rate (quantitative detection of λ-cyhalothrin within 8min), regeneration ability (maintaining good fluorescence properties after 8 cycling operation) and appreciable selectivity over several structural analogs. Moreover, the fluorescent sensor was further used to detect λ-cyhalothrin in real samples form the Beijing-Hangzhou Grand Canal Water. Despite the relatively complex components of the environmental water, the molecularly imprinted fluorescent hollow nanosensor still showed good recovery, clearly demonstrating the potential value of this smart sensor nanomaterial in environmental monitoring. Copyright © 2016 Elsevier B.V. All rights reserved.

The design and application of fluorophore–gold nanoparticle activatable probes

PubMed Central

Swierczewska, Magdalena; Lee, Seulki; Chen, Xiaoyuan

2013-01-01

Fluorescence-based assays and detection techniques are among the most highly sensitive and popular biological tests for researchers. To match the needs of research and the clinic, detection limits and specificities need to improve, however. One mechanism is to decrease non-specific background signals, which is most efficiently done by increasing fluorescence quenching abilities. Reports in the literature of theoretical and experimental work have shown that metallic gold surfaces and nanoparticles are ultra-efficient fluorescence quenchers. Based on these findings, subsequent reports have described gold nanoparticle fluorescence-based activatable probes that were designed to increase fluorescence intensity based on a range of stimuli. In this way, these probes can detect and signify assorted biomarkers and changes in environmental conditions. In this review, we explore the various factors and theoretical models that affect gold nanoparticle fluorescence quenching, explore current uses of activatable probes, and propose an engineering approach for future development of fluorescence based gold nanoparticle activatable probes. PMID:21380462

Signal amplification strategies for DNA and protein detection based on polymeric nanocomposites and polymerization: A review.

PubMed

Zhou, Shaohong; Yuan, Liang; Hua, Xin; Xu, Lingling; Liu, Songqin

2015-06-02

Demand is increasing for ultrasensitive bioassays for disease diagnosis, environmental monitoring and other research areas. This requires novel signal amplification strategies to maximize the signal output. In this review, we focus on a series of significant signal amplification strategies based on polymeric nanocomposites and polymerization. Some common polymers are used as carriers to increase the local concentration of signal probes and/or biomolecules on their surfaces or in their interiors. Some polymers with special fluorescence and optical properties can efficiently transfer the excitation energy from a single site to the whole polymer backbone. This results in superior fluorescence signal amplification due to the resulting collective effort (integration of signal). Recent polymerization-based signal amplification strategies that employ atom transfer radical polymerization (ATRP) and photo-initiated polymerization are also summarized. Several distinctive applications of polymers in ultrasensitive bioanalysis are highlighted. Copyright © 2015 Elsevier B.V. All rights reserved.

Efficient triplet harvesting of hybrid white organic light-emitting diodes using thermally activated delayed fluorescence green emitter

NASA Astrophysics Data System (ADS)

Lee, Song Eun; Lee, Ho Won; Baek, Hyun Jung; Yun, Tae Jun; Yun, Geum Jae; Kim, Woo Young; Kim, Young Kwan

2016-10-01

Hybrid white organic light-emitting diodes (WOLEDs) were fabricated by applying triplet harvesting (TH) using a green thermally activated delayed fluorescence (TADF) emitter. The triplet exciton of the green TADF emitter can be upconverted to its singlet state. The TH involved energy transfer of triplet exciton from a blue fluorescent emitter to a green TADF and red phosphorescent emitters, where they can decay radiatively. In addition, the triplet exciton of the green TADF emitter was energy transferred to its singlet state for a reverse intersystem crossing by green emission. Enhanced hybrid WOLEDs were demonstrated using an efficient green TADF emitter combined with red phosphorescent and blue fluorescent emitters. Hybrid WOLEDs were fabricated with various hole-electron recombination zones as changing blue emitting layer thicknesses. Among these, hybrid WOLEDs showed a maximum external quantum efficiency of 11.23%, luminous efficiency of 29.20 cd/A, and a power efficiency of 26.21 lm/W. Moreover, the WOLED exhibited electroluminescence spectra with Commission International de L'Éclairage chromaticity of (0.38, 0.36) at 1000 cd/m2 and a color rendering index of 82 at a practical brightness of 20,000 cd/m2.

Broadband magnetometry by infrared-absorption detection of nitrogen-vacancy ensembles in diamond

DOE Office of Scientific and Technical Information (OSTI.GOV)

Acosta, V. M.; Bauch, E.; Jarmola, A.

We demonstrate magnetometry by detection of the spin state of high-density nitrogen-vacancy ensembles in diamond using optical absorption at 1042 nm. With this technique, measurement contrast, and collection efficiency can approach unity, leading to an increase in magnetic sensitivity compared to the more common method of collecting red fluorescence. Working at 75 K with a sensor with effective volume 50x50x300 {mu}m{sup 3}, we project photon shot-noise limited sensitivity of 5 pT in one second of acquisition and bandwidth from dc to a few megahertz. Operation in a gradiometer configuration yields a noise floor of 7 nT{sub rms} at {approx}110 Hzmore » in one second of acquisition.« less

Novel multistep BRET-FRET energy transfer using nanoconjugates of firefly proteins, quantum dots, and red fluorescent proteins

NASA Astrophysics Data System (ADS)

Alam, Rabeka; Zylstra, Joshua; Fontaine, Danielle M.; Branchini, Bruce R.; Maye, Mathew M.

2013-05-01

Sequential bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET) from firefly luciferase to red fluorescent proteins using quantum dot or rod acceptor/donor linkers is described. The effect of morphology and tuned optical properties on the efficiency of this unique BRET-FRET system was evaluated.Sequential bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET) from firefly luciferase to red fluorescent proteins using quantum dot or rod acceptor/donor linkers is described. The effect of morphology and tuned optical properties on the efficiency of this unique BRET-FRET system was evaluated. Electronic supplementary information (ESI) available: Experimental details, Fig. S1 and Table S1-S4. See DOI: 10.1039/c3nr01842c

Design of Organic Solar Cells as a Function of Radiative Quantum Efficiency

NASA Astrophysics Data System (ADS)

Godefroid, Blaise; Kozyreff, Gregory

2017-09-01

We study the radiative decay, or fluorescence, of excitons in organic solar cells as a function of its geometrical parameters. Contrary to their nonradiative counterpart, fluorescence losses strongly depend on the environment. By properly tuning the thicknesses of the buffer layers between the active regions of the cell and the electrodes, the exciton lifetime and, hence, the exciton diffusion length can be increased. The importance of this phenomenon depends on the radiative quantum efficiency, which is the fraction of the exciton decay that is intrinsically due to fluorescence. Besides this effect, interferences within the cell control the efficiency of sunlight injection into the active layers. The optimal cell design must rely on a consideration of these two aspects. By properly managing fluorescence losses, one can significantly improve the cell performance. To demonstrate this fact, we use realistic material parameters inspired from literature data and obtain an increase of power-conversion efficiency from 11.3% to 12.7%. Conversely, not to take into account the strong dependence of fluorescence on the environment may lead to a suboptimal cell design and a degradation of cell performance. The presence of radiative losses, however small, significantly changes the optimal set of thicknesses. We illustrate the latter situation with experimental material data.

Single-frame 3D fluorescence microscopy with ultraminiature lensless FlatScope

PubMed Central

Adams, Jesse K.; Boominathan, Vivek; Avants, Benjamin W.; Vercosa, Daniel G.; Ye, Fan; Baraniuk, Richard G.; Robinson, Jacob T.; Veeraraghavan, Ashok

2017-01-01

Modern biology increasingly relies on fluorescence microscopy, which is driving demand for smaller, lighter, and cheaper microscopes. However, traditional microscope architectures suffer from a fundamental trade-off: As lenses become smaller, they must either collect less light or image a smaller field of view. To break this fundamental trade-off between device size and performance, we present a new concept for three-dimensional (3D) fluorescence imaging that replaces lenses with an optimized amplitude mask placed a few hundred micrometers above the sensor and an efficient algorithm that can convert a single frame of captured sensor data into high-resolution 3D images. The result is FlatScope: perhaps the world’s tiniest and lightest microscope. FlatScope is a lensless microscope that is scarcely larger than an image sensor (roughly 0.2 g in weight and less than 1 mm thick) and yet able to produce micrometer-resolution, high–frame rate, 3D fluorescence movies covering a total volume of several cubic millimeters. The ability of FlatScope to reconstruct full 3D images from a single frame of captured sensor data allows us to image 3D volumes roughly 40,000 times faster than a laser scanning confocal microscope while providing comparable resolution. We envision that this new flat fluorescence microscopy paradigm will lead to implantable endoscopes that minimize tissue damage, arrays of imagers that cover large areas, and bendable, flexible microscopes that conform to complex topographies. PMID:29226243

Spectral unmixing of multi-color tissue specific in vivo fluorescence in mice

NASA Astrophysics Data System (ADS)

Zacharakis, Giannis; Favicchio, Rosy; Garofalakis, Anikitos; Psycharakis, Stylianos; Mamalaki, Clio; Ripoll, Jorge

2007-07-01

Fluorescence Molecular Tomography (FMT) has emerged as a powerful tool for monitoring biological functions in vivo in small animals. It provides the means to determine volumetric images of fluorescent protein concentration by applying the principles of diffuse optical tomography. Using different probes tagged to different proteins or cells, different biological functions and pathways can be simultaneously imaged in the same subject. In this work we present a spectral unmixing algorithm capable of separating signal from different probes when combined with the tomographic imaging modality. We show results of two-color imaging when the algorithm is applied to separate fluorescence activity originating from phantoms containing two different fluorophores, namely CFSE and SNARF, with well separated emission spectra, as well as Dsred- and GFP-fused cells in F5-b10 transgenic mice in vivo. The same algorithm can furthermore be applied to tissue-specific spectroscopy data. Spectral analysis of a variety of organs from control, DsRed and GFP F5/B10 transgenic mice showed that fluorophore detection by optical systems is highly tissue-dependent. Spectral data collected from different organs can provide useful insight into experimental parameter optimisation (choice of filters, fluorophores, excitation wavelengths) and spectral unmixing can be applied to measure the tissue-dependency, thereby taking into account localized fluorophore efficiency. Summed up, tissue spectral unmixing can be used as criteria in choosing the most appropriate tissue targets as well as fluorescent markers for specific applications.

Application of CORSIKA Simulation Code to Study Lateral and Longitudinal Distribution of Fluorescence Light in Cosmic Ray Extensive Air Showers

NASA Astrophysics Data System (ADS)

Bagheri, Zahra; Davoudifar, Pantea; Rastegarzadeh, Gohar; Shayan, Milad

2017-03-01

In this paper, we used CORSIKA code to understand the characteristics of cosmic ray induced showers at extremely high energy as a function of energy, detector distance to shower axis, number, and density of secondary charged particles and the nature particle producing the shower. Based on the standard properties of the atmosphere, lateral and longitudinal development of the shower for photons and electrons has been investigated. Fluorescent light has been collected by the detector for protons, helium, oxygen, silicon, calcium and iron primary cosmic rays in different energies. So we have obtained a number of electrons per unit area, distance to the shower axis, shape function of particles density, percentage of fluorescent light, lateral distribution of energy dissipated in the atmosphere and visual field angle of detector as well as size of the shower image. We have also shown that location of highest percentage of fluorescence light is directly proportional to atomic number of elements. Also we have shown when the distance from shower axis increases and the shape function of particles density decreases severely. At the first stages of development, shower axis distance from detector is high and visual field angle is small; then with shower moving toward the Earth, angle increases. Overall, in higher energies, the fluorescent light method has more efficiency. The paper provides standard calibration lines for high energy showers which can be used to determine the nature of the particles.

Improved detection of soma location and morphology in fluorescence microscopy images of neurons.

PubMed

Kayasandik, Cihan Bilge; Labate, Demetrio

2016-12-01

Automated detection and segmentation of somas in fluorescent images of neurons is a major goal in quantitative studies of neuronal networks, including applications of high-content-screenings where it is required to quantify multiple morphological properties of neurons. Despite recent advances in image processing targeted to neurobiological applications, existing algorithms of soma detection are often unreliable, especially when processing fluorescence image stacks of neuronal cultures. In this paper, we introduce an innovative algorithm for the detection and extraction of somas in fluorescent images of networks of cultured neurons where somas and other structures exist in the same fluorescent channel. Our method relies on a new geometrical descriptor called Directional Ratio and a collection of multiscale orientable filters to quantify the level of local isotropy in an image. To optimize the application of this approach, we introduce a new construction of multiscale anisotropic filters that is implemented by separable convolution. Extensive numerical experiments using 2D and 3D confocal images show that our automated algorithm reliably detects somas, accurately segments them, and separates contiguous ones. We include a detailed comparison with state-of-the-art existing methods to demonstrate that our algorithm is extremely competitive in terms of accuracy, reliability and computational efficiency. Our algorithm will facilitate the development of automated platforms for high content neuron image processing. A Matlab code is released open-source and freely available to the scientific community. Copyright © 2016 Elsevier B.V. All rights reserved.

Energy Efficiency Comparison between Compact Fluorescent Lamp and Common Light Bulb

ERIC Educational Resources Information Center

Tanushevsk, Atanas; Rendevski, Stojan

2016-01-01

For acquainting the students of applied physics and students of teaching physics with the concept of energy efficiency, electrical and spectral characteristics of two widely used lamps--integrated fluorescence lamp and common light bulb have been investigated. Characterization of the lamps has been done by measuring the spectral irradiance and…

A wind tunnel test of newly developed personal bioaerosol samplers.

PubMed

Su, Wei-Chung; Tolchinsky, Alexander D; Sigaev, Vladimir I; Cheng, Yung Sung

2012-07-01

In this study the performance of two newly developed personal bioaerosol samplers was evaluated. The two test samplers are cyclone-based personal samplers that incorporate a recirculating liquid film. The performance evaluations focused on the physical efficiencies that a personal bioaerosol sampler could provide, including aspiration, collection, and capture efficiencies. The evaluation tests were carried out in a wind tunnel, and the test personal samplers were mounted on the chest of a full-size manikin placed in the test chamber of the wind tunnel. Monodisperse fluorescent aerosols ranging from 0.5 to 20 microm were used to challenge the samplers. Two wind speeds of 0.5 and 2.0 m/sec were employed as the test wind speeds in this study. The test results indicated that the aspiration efficiency of the two test samplers closely agreed with the ACGIH inhalable convention within the size range of the test aerosols. The aspiration efficiency was found to be independent of the sampling orientation. The collection efficiency acquired from these two samplers showed that the 50% cutoff diameters were both around 0.6 microm. However the wall loss of these two test samplers increased as the aerosol size increased, and the wall loss of PAS-4 was considerably higher than that of PAS-5, especially in the aerosol size larger than 5 microm, which resulted in PAS-4 having a relatively lower capture efficiency than PAS-5. Overall, the PAS-5 is considered a better personal bioaerosol sampler than the PAS-4.

Shape Engineering Boosts Magnetic Mesoporous Silica Nanoparticle-Based Isolation and Detection of Circulating Tumor Cells.

PubMed

Chang, Zhi-Min; Wang, Zheng; Shao, Dan; Yue, Juan; Xing, Hao; Li, Li; Ge, Mingfeng; Li, Mingqiang; Yan, Huize; Hu, Hanze; Xu, Qiaobing; Dong, Wen-Fei

2018-04-04

Magnetic mesoporous silica nanoparticles (M-MSNs) are attractive candidates for the immunomagnetic isolation and detection of circulating tumor cells (CTCs). Understanding of the interactions between the effects of the shape of M-MSNs and CTCs is crucial to maximize the binding capacity and capture efficiency as well as to facilitate the sensitivity and efficiency of detection. In this work, fluorescent M-MSNs were rationally designed with sphere and rod morphologies while retaining their robust fluorescence and uniform surface functionality. After conjugation with the antibody of epithelial cell adhesion molecule (EpCAM), both of the differently shaped M-MSNs-EpCAM obtained achieved efficient enrichment of CTCs and fluorescent-based detection. Importantly, rodlike M-MSNs exhibited faster immunomagnetic isolation as well as better performance in the isolation and detection of CTCs in spiked cells and real clinical blood samples than those of their spherelike counterparts. Our results showed that shape engineering contributes positively toward immunomagnetic isolation, which might open new avenues to the rational design of magnetic-fluorescent nanoprobes for the sensitive and efficient isolation and detection of CTCs.

Quantification issues of trace metal contaminants on silicon wafers by means of TOF-SIMS, ICP-MS, and TXRF

NASA Astrophysics Data System (ADS)

Rostam-Khani, P.; Hopstaken, M. J. P.; Vullings, P.; Noij, G.; O'Halloran, O.; Claassen, W.

2004-06-01

Measurement of surface metal contamination on silicon wafers is essential for yield enhancement in IC manufacturing. Vapor phase decomposition coupled with either inductively coupled plasma mass spectrometry (VPD-ICP-MS), or total reflection X-ray fluorescence (VPD-TXRF), TXRF and more recently time of flight secondary ion mass spectrometry (TOF-SIMS) are used to monitor surface metal contamination. These techniques complement each other in their respective strengths and weaknesses. For reliable and accurate quantification, so-called relative sensitivity factors (RSF) are required for TOF-SIMS analysis. For quantification purposes in VPD, the collection efficiency (CE) is important to ensure complete collection of contamination. A standard procedure has been developed that combines the determination of these RSFs as well as the collection efficiency using all the analytical techniques mentioned above. Therefore, sample wafers were intentionally contaminated and analyzed (by TOF-SIMS) directly after preparation. After VPD-ICP-MS, several scanned surfaces were analyzed again by TOF-SIMS. Comparing the intensities of the specific metals before and after the VPD-DC procedure on the scanned surface allows the determination of so-called removing efficiency (RE). In general, very good agreement was obtained comparing the four analytical techniques after updating the RSFs for TOF-SIMS. Progress has been achieved concerning the CE evaluation as well as determining the RSFs more precisely for TOF-SIMS.

Rare Earth Fluorescent Nanomaterials for Enhanced Development of Latent Fingerprints.

PubMed

Wang, Meng; Li, Ming; Yu, Aoyang; Wu, Jian; Mao, Chuanbin

2015-12-30

The most commonly found fingerprints at crime scenes are latent and, thus, an efficient method for detecting latent fingerprints is very important. However, traditional developing techniques have drawbacks such as low developing sensitivity, high background interference, complicated operation, and high toxicity. To tackle this challenge, we have synthesized two kinds of rare earth fluorescent nanomaterials, including the fluoresce red-emitting YVO4:Eu nanocrystals and green-emitting LaPO4:Ce,Tb nanobelts, and then used them as fluorescent labels for the development of latent fingerprints with high sensitivity, high contrast, high selectivity, high efficiency, and low background interference, on various substrates including noninfiltrating materials, semi-infiltrating materials, and infiltrating materials.

[Prokaryotic community of subglacial bottom sediments of Antarctic Lake Untersee: detection by cultural and direct microscopic techniques].

PubMed

Muliukin, A L; Demkina, E V; Manucharova, N A; Akimov, V N; Andersen, D; McKay, C; Gal'chenko, V F

2014-01-01

The heterotrophic mesophilic component was studied in microbial communities of the samples of frozen regolith collected from the glacier near Lake Untersee collected in 2011 during the joint Russian-American expedition to central Dronning Maud Land (Eastern Antarctica). Cultural techniques revealed high bacterial numbers in the samples. For enumeration of viable cells, the most probable numbers (MPN) method proved more efficient than plating on agar media. Fluorescent in situ hybridization with the relevant oligonucleotide probes revealed members of the groups Eubacteria (Actinobacteria, Firmicutes) and Archaea. Application of the methods of cell resuscitation, such as the use of diluted media and prevention of oxidative stress, did not result in a significant increase in the numbers of viable cells retrieved form subglacial sediment samples. Our previous investigations demonstrated the necessity for special procedures for efficient reactivation of the cells from microbial communities of preserved fossil soil and permafrost samples collected in the Arctic zone. The differences in response to the special resuscitation procedures may reflect the differences in the physiological and morphological state of bacterial cells in microbial communities subject to continuous or periodic low temperatures and dehydration.

Development of a Technique for Separating Raman Scattering Signals from Background Emission with Single-Shot Measurement Potential

NASA Technical Reports Server (NTRS)

Hartfield, Roy

1996-01-01

Raman scattering is a powerful technique for quantitatively probing high temperature and high speed flows. However, this technique has typically been limited to clean hydrogen flames because of the broadband fluorescence interference which occurs in hydrocarbon flames. Fluorescence can also interfere with the Raman signal in clean hydrogen flames when broadband UV lasers are used as the scattering source. A solution to this problem has been demonstrated. The solution to the fluorescence interference lies in the fact that the vibrational Q-branch Raman signal is highly polarized for 90 deg. signal collection and the fluorescence background is essentially unpolarized. Two basic schemes are available for separating the Raman from the background. One scheme involves using a polarized laser and collecting a signal with both horizontal and vertical laser polarizations separately. The signal with the vertical polarization will contain both the Raman and the fluorescence while the signal with the horizontal polarization will contain only the fluorescence. The second scheme involves polarization discrimination on the collection side of the optical setup. For vertical laser polarization, the scattered Q-branch Raman signal will be vertically polarized; hence the two polarizations can be collected separately and the difference between the two is the Raman signal. This approach has been used for the work found herein and has the advantage of allowing the data to be collected from the same laser shot(s). This makes it possible to collect quantitative Raman data with single shot resolution in conditions where interference cannot otherwise be eliminated.

High efficiency and brightness fluorescent organic light emitting diode by triplet-triplet fusion

DOEpatents

Forrest, Stephen; Zhang, Yifan

2015-02-10

A first device is provided. The first device further comprises an organic light emitting device. The organic light emitting device further comprises an anode, a cathode, and an emissive layer disposed between the anode and the cathode. The emissive layer may include an organic host compound and at least one organic emitting compound capable of fluorescent emission at room temperature. Various configurations are described for providing a range of current densities in which T-T fusion dominates over S-T annihilation, leading to very high efficiency fluorescent OLEDs.

Fluorescence imaging of the nanoparticles modified with indocyanine green

NASA Astrophysics Data System (ADS)

Gareev, K. G.; Babikova, K. Y.; Postnov, V. N.; Naumisheva, E. B.; Korolev, D. V.

2017-11-01

The comparative research of silica, the magnetite and magnetite-silica nanoparticles modified with fluorescent dyes using gas-phase and liquid-phase methods was conducted. At the content of fluorescent dye comparable in size a particular spectrophotometric method, nanoparticles with fluorescein have up to 1000 times larger overall luminous efficiency. It is revealed that magnetic nanoparticles are characterized by a smaller light efficiency in comparison with silica particles, at the same time particles of a magnetite are most effective at modification with fluorescein, and magnetite-silica particles - at modification with indocyanine green.

Efficient fluorescence/phosphorescence white organic light-emitting diodes with ultra high color stability and mild efficiency roll-off

NASA Astrophysics Data System (ADS)

Du, Xiaoyang; Tao, Silu; Huang, Yun; Yang, Xiaoxia; Ding, Xulin; Zhang, Xiaohong

2015-11-01

Efficient fluorescence/phosphorescence hybrid white organic light-emitting diodes (OLEDs) with single doped co-host structure have been fabricated. Device using 9-Naphthyl-10 -(4-triphenylamine)anthrancene as the fluorescent dopant and Ir(ppy)3 and Ir(2-phq)3 as the green and orange phosphorescent dopants show the luminous efficiency of 12.4% (17.6 lm/W, 27.5 cd/A) at 1000 cd/m2. Most important to note that the efficiency-brightness roll-off of the device was very mild. With the brightness rising up to 5000 and 10 000 cd/m2, the efficiency could be kept at 11.8% (14.0 lm/W, 26.5 cd/A) and 11.0% (11.8 lm/W, 25.0 cd/A). The Commission Internationale de L'Eclairage (CIE) coordinates and color rending index (CRI) were measured to be (0.45, 0.48) and 65, respectively, and remained the same in a large range of brightness (1000-10 000 cd/m2), which is scarce in the reported white OLEDs. The performance of the device at high luminance (5000 and 10 000 cd/m2) was among the best reported results including fluorescence/phosphorescence hybrid and all-phosphorescent white OLEDs. Moreover, the CRI of the white OLED can be improved to 83 by using a yellow-green emitter (Ir(ppy)2bop) in the device.

Efficient genome editing by FACS enrichment of paired D10A Cas9 nickases coupled with fluorescent proteins.

PubMed

Gopalappa, Ramu; Song, Myungjae; Chandrasekaran, Arun Pandian; Das, Soumyadip; Haq, Saba; Koh, Hyun Chul; Ramakrishna, Suresh

2018-05-31

Targeted genome editing by clustered regularly interspaced short palindromic repeats (CRISPR-Cas9) raised concerns over off-target effects. The use of double-nicking strategy using paired Cas9 nickase has been developed to minimize off-target effects. However, it was reported that the efficiency of paired nickases were comparable or lower than that of either corresponding nuclease alone. Recently, we conducted a systematic comparison of the efficiencies of several paired Cas9 with their corresponding Cas9 nucleases and showed that paired D10A Cas9 nickases are sometimes more efficient than individual nucleases for gene disruption. However, sometimes the designed paired Cas9 nickases exhibited significantly lower mutation frequencies than nucleases, hampering the generation of cells containing paired Cas9 nickase-induced mutations. Here we implemented IRES peptide-conjugation of fluorescent protein to Cas9 nickase and subjected for fluorescence-activated cell sorting. The sorted cell populations are highly enriched with cells containing paired Cas9 nickase-induced mutations, by a factor of up to 40-fold as compared with the unsorted population. Furthermore, gene-disrupted single cell clones using paired nickases followed by FACS sorting strategy were generated highly efficiently, without compromising with its low off-target effects. We envision that our fluorescent protein coupled paired nickase-mediated gene disruption, facilitating efficient and highly specific genome editing in medical research.

A plasmid collection for PCR-based gene targeting in the filamentous ascomycete Ashbya gossypii.

PubMed

Kaufmann, Andreas

2009-08-01

PCR-based gene targeting with heterologous markers is an efficient method to delete genes, generate gene fusions, and modulate gene expression. For the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, several plasmid collections are available covering a wide range of tags and markers. For several reasons, many of these cassettes cannot be used in the filamentous ascomycete Ashbya gossypii. This article describes the construction of 93 heterologous modules for C- and N-terminal tagging and promoter replacements in A. gossypii. The performance of 12 different fluorescent tags was evaluated by monitoring their brightness, detectability, and photostability when fused to the myosin light-chain protein Mlc2. Furthermore, the thiamine-repressible S. cerevisiae THI13 promoter was established to regulate gene expression in A. gossypii. This collection will help accelerate analysis of gene function in A. gossypii and in other ascomycetes where S. cerevisiae promoter elements are functional.

Rapid Global Fitting of Large Fluorescence Lifetime Imaging Microscopy Datasets

PubMed Central

Warren, Sean C.; Margineanu, Anca; Alibhai, Dominic; Kelly, Douglas J.; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Katan, Matilda

2013-01-01

Fluorescence lifetime imaging (FLIM) is widely applied to obtain quantitative information from fluorescence signals, particularly using Förster Resonant Energy Transfer (FRET) measurements to map, for example, protein-protein interactions. Extracting FRET efficiencies or population fractions typically entails fitting data to complex fluorescence decay models but such experiments are frequently photon constrained, particularly for live cell or in vivo imaging, and this leads to unacceptable errors when analysing data on a pixel-wise basis. Lifetimes and population fractions may, however, be more robustly extracted using global analysis to simultaneously fit the fluorescence decay data of all pixels in an image or dataset to a multi-exponential model under the assumption that the lifetime components are invariant across the image (dataset). This approach is often considered to be prohibitively slow and/or computationally expensive but we present here a computationally efficient global analysis algorithm for the analysis of time-correlated single photon counting (TCSPC) or time-gated FLIM data based on variable projection. It makes efficient use of both computer processor and memory resources, requiring less than a minute to analyse time series and multiwell plate datasets with hundreds of FLIM images on standard personal computers. This lifetime analysis takes account of repetitive excitation, including fluorescence photons excited by earlier pulses contributing to the fit, and is able to accommodate time-varying backgrounds and instrument response functions. We demonstrate that this global approach allows us to readily fit time-resolved fluorescence data to complex models including a four-exponential model of a FRET system, for which the FRET efficiencies of the two species of a bi-exponential donor are linked, and polarisation-resolved lifetime data, where a fluorescence intensity and bi-exponential anisotropy decay model is applied to the analysis of live cell homo-FRET data. A software package implementing this algorithm, FLIMfit, is available under an open source licence through the Open Microscopy Environment. PMID:23940626

PHOTOSYNTHETIC EFFICIENCY OF MARINE PLANTS

PubMed Central

Yocum, C. S.; Blinks, L. R.

1954-01-01

Multicellular marine plants were collected from their natural habitats and the quantum efficiency of their photosynthesis was determined in the laboratory in five narrow wave length bands in the visible spectrum. The results along with estimates of the relative absorption by the various plastid pigments show a fairly uniform efficiency of 0.08 molecules O2 per absorbed quantum for (a) chlorophyll of one flowering plant, green algae, and brown algae, (b) fucoxanthol and other carotenoids of brown algae, and (c) the phycobilin pigments phycocyanin and phycoerythrin of red algae. The carotenoids of green algae are sometimes less efficient while those of red algae are largely or entirely inactive. Chlorophyll a of red algae is about one-half as efficient (φo2 = 0.04) as either the phycobilins, or the chlorophyll of most other plants. These results as well as those of high intensity and of fluorescence experiments are consistent with a mechanism in which about half the chlorophyll is inactive while the other half is fully active and is an intermediate in phycoerythrin- and phycocyanin-sensitized photosynthesis. PMID:13192311

Using simulated fluorescence cell micrographs for the evaluation of cell image segmentation algorithms.

PubMed

Wiesmann, Veit; Bergler, Matthias; Palmisano, Ralf; Prinzen, Martin; Franz, Daniela; Wittenberg, Thomas

2017-03-18

Manual assessment and evaluation of fluorescent micrograph cell experiments is time-consuming and tedious. Automated segmentation pipelines can ensure efficient and reproducible evaluation and analysis with constant high quality for all images of an experiment. Such cell segmentation approaches are usually validated and rated in comparison to manually annotated micrographs. Nevertheless, manual annotations are prone to errors and display inter- and intra-observer variability which influence the validation results of automated cell segmentation pipelines. We present a new approach to simulate fluorescent cell micrographs that provides an objective ground truth for the validation of cell segmentation methods. The cell simulation was evaluated twofold: (1) An expert observer study shows that the proposed approach generates realistic fluorescent cell micrograph simulations. (2) An automated segmentation pipeline on the simulated fluorescent cell micrographs reproduces segmentation performances of that pipeline on real fluorescent cell micrographs. The proposed simulation approach produces realistic fluorescent cell micrographs with corresponding ground truth. The simulated data is suited to evaluate image segmentation pipelines more efficiently and reproducibly than it is possible on manually annotated real micrographs.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bazalova, M; Ahmad, M; Fahrig, R

Purpose: To evaluate x-ray fluorescence computed tomography induced with proton beams (pXFCT) for imaging of gold contrast agent. Methods: Proton-induced x-ray fluorescence was studied by means of Monte Carlo (MC) simulations using TOPAS, a MC code based on GEANT4. First, proton-induced K-shell and L-shell fluorescence was studied as a function of proton beam energy and 1) depth in water and 2) size of contrast object. Second, pXFCT images of a 2-cm diameter cylindrical phantom with four 5- mm diameter contrast vials and of a 20-cm diameter phantom with 1-cm diameter vials were simulated. Contrast vials were filled with water andmore » water solutions with 1-5% gold per weight. Proton beam energies were varied from 70-250MeV. pXFCT sinograms were generated based on the net number of gold K-shell or L-shell x-rays determined by interpolations from the neighboring 0.5keV energy bins of spectra collected with an idealized 4π detector. pXFCT images were reconstructed with filtered-back projection, and no attenuation correction was applied. Results: Proton induced x-ray fluorescence spectra showed very low background compared to x-ray induced fluorescence. Proton induced L-shell fluorescence had a higher cross-section compared to K-shell fluorescence. Excitation of L-shell fluorescence was most efficient for low-energy protons, i.e. at the Bragg peak. K-shell fluorescence increased with increasing proton beam energy and object size. The 2% and 5% gold contrast vials were accurately reconstructed in K-shell pXFCT images of both the 2-cm and 20-cm diameter phantoms. Small phantom L-shell pXFCT image required attenuation correction and had a higher sensitivity for 70MeV protons compared to 250MeV protons. With attenuation correction, L-shell pXFCT might be a feasible option for imaging of small size (∼2cm) objects. Imaging doses for all simulations were 5-30cGy. Conclusion: Proton induced x-ray fluorescence CT promises to be an alternative quantitative imaging technique to the commonly considered XFCT imaging with x-ray beams.« less

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, Alexander M.; Benson, Scott C.

1999-01-01

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, Alexander M.; Benson, Scott C.

1998-01-01

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, Alexander N.; Benson, Scott C.

1995-01-01

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, Alexander N.; Benson, Scott C.

1997-01-01

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Efficiency and concentration ratio measurements of fluorescent solar concentrators using a xenon measurement system.

PubMed

Heidler, K

1981-03-01

An indoor test stand for fluorescent planar concentrator-collectors (FPC) with a 1.6-kW xenon light source, irradiating a rectangular triangle (active area 400 cm(2)) with 82 mW cm(-2), and a typical homogeneity of +/-10% is described in detail. Conditions and definitions for the efficiency measurements are discussed, taking intensity and reflection effects into account. As an illustration FPC efficiency and concentration ratios are reported (best optical efficiency was 7.5% for a single-plate one-dye FPC), the influence of thickness variation is discussed, and an enhancement in solar cell efficiency is demonstrated.

Detection of Mycobacterium avium subsp. paratuberculosis in bovine manure using Whatman FTA card technology and Lightcycler real-time PCR.

PubMed

Jaravata, Carmela V; Smith, Wayne L; Rensen, Gabriel J; Ruzante, Juliana M; Cullor, James S

2006-01-01

A modified forensic DNA extraction and real-time fluorescent polymerase chain reaction assay has been evaluated for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine fecal samples using primers and fluorescent resonance energy transfer (FRET) probes targeting the IS900 gene sequence of MAP. DNA was successfully extracted from manure samples by utilizing the Whatman FTA card technology, which allows for simple processing and storage of samples at room temperature. The FTA cards were washed and subjected to a Chelex-100 incubation to remove any remaining polymerase chain reaction (PCR) inhibitors and to elute the DNA from the FTA card. This isolated DNA was then subjected to direct real time fluorescent PCR analysis. Detection of MAP DNA from bovine fecal samples spiked with known concentrations of viable MAP cells was obtained. The detection limits of the assay was consistently found to be between 10(2) and 10(4) colony forming units [CFU]/g, with some samples containing as low as 10 CFU/g, yielding positive assay results. This cost-efficient assay allows reporting of results as early as 4 h after fecal collection, which can be particularly useful in highthroughput herd screening.

Estimating background-subtracted fluorescence transients in calcium imaging experiments: a quantitative approach.

PubMed

Joucla, Sébastien; Franconville, Romain; Pippow, Andreas; Kloppenburg, Peter; Pouzat, Christophe

2013-08-01

Calcium imaging has become a routine technique in neuroscience for subcellular to network level investigations. The fast progresses in the development of new indicators and imaging techniques call for dedicated reliable analysis methods. In particular, efficient and quantitative background fluorescence subtraction routines would be beneficial to most of the calcium imaging research field. A background-subtracted fluorescence transients estimation method that does not require any independent background measurement is therefore developed. This method is based on a fluorescence model fitted to single-trial data using a classical nonlinear regression approach. The model includes an appropriate probabilistic description of the acquisition system's noise leading to accurate confidence intervals on all quantities of interest (background fluorescence, normalized background-subtracted fluorescence time course) when background fluorescence is homogeneous. An automatic procedure detecting background inhomogeneities inside the region of interest is also developed and is shown to be efficient on simulated data. The implementation and performances of the proposed method on experimental recordings from the mouse hypothalamus are presented in details. This method, which applies to both single-cell and bulk-stained tissues recordings, should help improving the statistical comparison of fluorescence calcium signals between experiments and studies. Copyright © 2013 Elsevier Ltd. All rights reserved.

Contrast Induced by a Static Magnetic Field for Improved Detection in Nanodiamond Fluorescence Microscopy

NASA Astrophysics Data System (ADS)

Singam, Shashi K. R.; Motylewski, Jaroslaw; Monaco, Antonina; Gjorgievska, Elena; Bourgeois, Emilie; Nesládek, Milos; Giugliano, Michele; Goovaerts, Etienne

2016-12-01

Diamond nanoparticles with negatively charged nitrogen-vacancy (NV) centers are highly efficient nonblinking emitters that exhibit spin-dependent intensity. An attractive application of these emitters is background-free fluorescence microscopy exploiting the fluorescence quenching induced either by resonant microwaves (RMWs) or by an applied static magnetic field (SMF). Here, we compare RMW- and SMF-induced contrast measurements over a wide range of optical excitation rates for fluorescent nanodiamonds (FNDs) and for NV centers shallowly buried under the (100)-oriented surface of a diamond single crystal (SC). Contrast levels are found to be systematically lower in the FNDs than in the SC. At low excitation rates, the RMW contrast initially rises to a maximum (up to 7% in FNDs and 13% in the SC) but then decreases steadily at higher intensities. Conversely, the SMF contrast increases from approximately 12% at low excitation rates to high values of 20% and 38% for the FNDs and SC, respectively. These observations are well described in a rate-equations model for the charged NV defect using parameters in good agreement with the literature. The SMF approach yields higher induced contrast in image collection under commonly applied optical excitation. Unlike the RMW method, there is no thermal load exerted on the aqueous media in biological samples in the SMF approach. We demonstrate imaging by SMF-induced contrast in neuronal cultures incorporating FNDs (i) in a setup for patch-clamp experiments in parallel with differential-interference-contrast microscopy, (ii) after a commonly used staining procedure as an illustration of the high selectivity against background fluorescence, and (iii) in a confocal fluorescence microscope in combination with bright-field microscopy.

Near real time, accurate, and sensitive microbiological safety monitoring using an all-fibre spectroscopic fluorescence system

NASA Astrophysics Data System (ADS)

Vanholsbeeck, F.; Swift, S.; Cheng, M.; Bogomolny, E.

2013-11-01

Enumeration of microorganisms is an essential microbiological task for many industrial sectors and research fields. Various tests for detection and counting of microorganisms are used today. However most of the current methods to enumerate bacteria require either long incubation time for limited accuracy, or use complicated protocols along with bulky equipment. We have developed an accurate, all-fibre spectroscopic system to measure fluorescence signal in-situ. In this paper, we examine the potential of this setup for near real time bacteria enumeration in aquatic environment. The concept is based on a well-known phenomenon that the fluorescence quantum yields of some nucleic acid stains significantly increase upon binding with nucleic acids of microorganisms. In addition we have used GFP labeled organisms. The fluorescence signal increase can be correlated to the amount of nucleic acid present in the sample. In addition we have used GFP labeled organisms. Our results show that we are able to detect a wide range of bacteria concentrations without dilution or filtration (1-108 CFU/ml) using different optical probes we designed. This high sensitivity is due to efficient light delivery with an appropriate collection volume and in situ fluorescence detection as well as the use of a sensitive CCD spectrometer. By monitoring the laser power, we can account for laser fluctuations while measuring the fluorescence signal which improves as well the system accuracy. A synchronized laser shutter allows us to achieve a high SNR with minimal integration time, thereby reducing the photobleaching effect. In summary, we conclude that our optical setup may offer a robust method for near real time bacterial detection in aquatic environment.

Energy efficient fluorescent ballasts. Phase I, final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stevens Luminoptics Corporation

1978-06-21

The development of a high-frequency electronic (Stevens) ballast for fluorescent lamps is described. It is claimed that use of this ballast could reduce use energy consumption by 1.2 to 2.5 percent. The Stevens ballast has a basic efficiency of 29 percent when used with conventional lamps. With the more efficient lamps, the efficiency increases drastically. The conventional ballast and lamp has an efficiacy of approximately 60 to 63 lumens per watt (LPW). With the Stevens ballast the efficiacy raises to between 75 and 80 lumens per watt. When the Stevens ballast is utilized with the newer high efficiency lamps themore » efficiacy increases to 90 to 95 lumens per watt or a full 51 percent improvement over conventional coil and core ballasts and 25 percent over the best high efficiency premium coil and core ballasts. In addition to its energy savings capabilities, this high frequency fluorescent lamp ballast has the advantages that it is a true retrofit device that is directly interchangeable with the conventional coil core ballast, and it is dimmable over a wide and continuous range. (LCLC)« less

Real-time and quantitative fluorescent live-cell imaging with quadruplex-specific red-edge probe (G4-REP).

PubMed

Yang, Sunny Y; Amor, Souheila; Laguerre, Aurélien; Wong, Judy M Y; Monchaud, David

2017-05-01

The development of quadruplex-directed molecular diagnostic and therapy rely on mechanistic insights gained at both cellular and tissue levels by fluorescence imaging. This technique is based on fluorescent reporters that label cellular DNA and RNA quadruplexes to spatiotemporally address their complex cell biology. The photophysical characteristics of quadruplex probes usually dictate the modality of cell imaging by governing the selection of the light source (lamp, LED, laser), the optical light filters and the detection modality. Here, we report the characterizations of prototype from a new generation of quadruplex dye termed G4-REP (for quadruplex-specific red-edge probe) that provides fluorescence responses regardless of the excitation wavelength and modality (owing to the versatility gained through the red-edge effect), thus allowing for diverse applications and most imaging facilities. This is demonstrated by cell images (and associated quantifications) collected through confocal and multiphoton microscopy as well as through real-time live-cell imaging system over extended period, monitoring both non-cancerous and cancerous human cell lines. Our results promote a new way of designing versatile, efficient and convenient quadruplex-reporting dyes for tracking these higher-order nucleic acid structures in living human cells. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio. Copyright © 2016 Elsevier B.V. All rights reserved.

User-Friendly End Station at the ALS for Nanostructure Characterization

DOE Office of Scientific and Technical Information (OSTI.GOV)

F. J. Himpsel; P. Alivisatos; T. Callcott

2006-07-05

This is a construction project for an end station at the ALS, which is optimized for measuring NEXAFS of nanostructures with fluorescence detection. Compared to the usual electron yield detection, fluorescence is able to probe buried structures and is sensitive to dilute species, such as nanostructures supported on a substrate. Since the quantum yield for fluorescence is 10{sup -4}-10{sup -5} times smaller than for electrons in the soft x-ray regime, such an end station requires bright undulator beamlines at the ALS. In order to optimize the setup for a wide range of applications, two end stations were built: (1) Amore » simple, mobile chamber with efficient photon detection (>10{sup 4} times the solid angle collection of fluorescence spectrographs) and a built-in magnet for MCD measurements at EPU beamlines (Fig. 1 left). It allows rapid mapping the electronic states of nanostructures (nanocrystals, nanowires, tailored magnetic materials, buried interfaces, biologically-functionalized surfaces). It was used with BL 8.0 (linear polarized undulator) and BL 4.0 (variable polarization). (2) A sophisticated, stationary end station operating at Beamline 8.0 (Fig. 1 right). It contains an array of surface characterization instruments and a micro-focus capability for scanning across graded samples (wedges for thickness variation, stoichiometry gradients, and general variations of the sample preparation conditions for optimizing nanostructures).« less

Characterisation and qualification of natural organic matter with a new online fluorescene sensor

NASA Astrophysics Data System (ADS)

Wagner, Martin; Dahlaus, Anna; Moldaenke, Christian; Schmidt, Wido

2016-04-01

Natural organic water compounds are determined usually with the bulk parameter DOC (dissolved organic carbon). The DOC is a heterogeneous parameter which consists of various organic fractions and shows often spatially as well as temporally a high dynamic range. The fluorescence spectroscopy is a tool for measuring individual DOC groups in a quick and easy way. A fluorescence sensor was developed within the framework of a research project that provides online detection of humic substances and organic polymers. Humic substances can be differentiated fulvic and humic acids, bio-polymers in proteins and algal chlorophyll-a. The chlorophyll fluorescence can be additionally assigned to green algae and diatoms as well as in cyanobacteria. The sensor has been tested during several measurement programs and was used in various waterworks for monitoring of raw water and treated water. The sensor is based on LED technology, works long term stable and is of low maintenance due to an autonomous cleaning unit. Using the sensor qualitative and quantitative changes of the raw water during drinking water treatment could be estimated efficiently. The processing stage of flocculation/filtration showed a significant reduction in the humic substances concentration, where macromolecular humic acids were removed with higher efficiency than low molecular weighted fulvic acids. Dynamical, seasonal-related processes in the water body of a drinking water reservoir could also be traced. Seasonal and single-event-related changes in temperature, turbidity and the composition of humic substances and algae were collected with high sensitivity for example during the autumn circulation in the water body.

Fluorescent pyrimidine ribonucleotide: synthesis, enzymatic incorporation and utilization

PubMed Central

Srivatsan, Seergazhi G.

2008-01-01

Fluorescent nucleobase analogs that respond to changes in their microenvironment are valuable for studying RNA structure, dynamics and recognition. The most commonly used fluorescent ribonucleoside is 2-aminopurine, a highly responsive purine analog. Responsive isosteric fluorescent pyrimidine analogs are, however, rare. Appending 5-membered aromatic heterocycles at the 5-position on a pyrimidine core has recently been found to provide a family of responsive fluorescent nucleoside analogs with emission in the visible range. To explore the potential utility of this chromophore for studying RNA–ligand interactions, an efficient incorporation method is necessary. Here we describe the synthesis of the furan-containing ribonucleoside and its triphosphate, as well as their basic photophysical characteristics. We demonstrate that T7 RNA polymerase accepts this fluorescent ribonucleoside triphosphate as a substrate in in vitro transcription reactions and very efficiently incorporates it into RNA oligonucleotides, generating fluorescent constructs. Furthermore, we utilize this triphosphate for the enzymatic preparation of a fluorescent bacterial A-site, an RNA construct of potential therapeutic utility. We show that the binding of this RNA target to aminoglycoside antibiotics, its cognate ligands, can be effectively monitored by fluorescence spectroscopy. These observations are significant since isosteric emissive U derivatives are scarce and the trivial synthesis and effective enzymatic incorporation of the furan-containing U triphosphate make it accessible to the biophysical community. PMID:17256858

Enhanced speed in fluorescence imaging using beat frequency multiplexing

NASA Astrophysics Data System (ADS)

Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

2016-03-01

Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

New anthracene derivatives as triplet acceptors for efficient green-to-blue low-power upconversion.

PubMed

Liang, Zuo-Qin; Sun, Bin; Ye, Chang-Qing; Wang, Xiao-Mei; Tao, Xu-Tang; Wang, Qin-Hua; Ding, Ping; Wang, Bao; Wang, Jing-Jing

2013-10-21

Three new anthracene derivatives [2-chloro-9,10-dip-tolylanthracene (DTACl), 9,10-dip-tolylanthracene-2-carbonitrile (DTACN), and 9,10-di(naphthalen-1-yl)anthracene-2-carbonitrile (DNACN)] were synthesized as triplet acceptors for low-power upconversion. Their linear absorption, single-photon-excited fluorescence, and upconversion fluorescence properties were studied. The acceptors exhibit high fluorescence yields in DMF. Selective excitation of the sensitizer Pd(II)octaethylporphyrin (PdOEP) in solution containing DTACl, DTACN, or DNA-CN at 532 nm with an ultralow excitation power density of 0.5 W cm(-2) results in anti-Stokes blue emission. The maximum upconversion quantum yield (Φ(UC) =17.4%) was obtained for the couple PdOEP/DTACl. In addition, the efficiency of the triplet-triplet energy transfer process was quantitatively studied by quenching experiments. Experimental results revealed that a highly effective acceptor for upconversion should combine high fluorescence quantum yields with efficient quenching of the sensitizer triplet. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescent vancomycin and terephthalate comodified europium-doped layered double hydroxides nanoparticles: synthesis and application for bacteria labelling

NASA Astrophysics Data System (ADS)

Sun, Jianchao; Fan, Hai; Wang, Nan; Ai, Shiyun

2014-09-01

Vancomycin (Van)- and terephthalate (TA)-comodified europium-doped layered double hydroxides (Van-TA-Eu-LDHs) nanoparticles were successfully prepared by a two-step method, in which, TA acted as a sensitizer to enhance the fluorescent property and Van was modified on the surface of LDH to act as an affinity reagent to bacteria. The obtained products were characterized by X-ray diffraction, transmission electron microscope and fluorescent spectroscopy. The results demonstrated that the prepared Van- and TA-comodified europium-doped layered double hydroxides (Van-TA-Eu-LDHs) nanoparticles with diameter of 50 nm in size showed highly efficient fluorescent property. Furthermore, due to the high affinity of Van to bacteria, the prepared Van-TA-Eu-LDHs nanoparticles showed efficient bacteria labelling by fluorescent property. The prepared nanoparticles may have wide applications in the biological fields, such as biomolecular labelling and cell imaging.

Removal of fluorescent dissolved organic matter in biologically treated textile effluents by NDMP anion exchange process: efficiency and mechanism.

PubMed

Li, Wen-Tao; Xu, Zi-Xiao; Shuang, Chen-Dong; Zhou, Qing; Li, Hai-Bo; Li, Ai-Min

2016-03-01

The efficiency and mechanism of anion exchange resin Nanda Magnetic Polymer (NDMP) for removal of fluorescent dissolved organic matter in biologically treated textile effluents were studied. The bench-scale experiments showed that as well as activated carbon, anion exchange resin could efficiently remove both aniline-like and humic-like fluorescent components, which can be up to 40 % of dissolved organic matter. The humic-like fluorescent component HS-Em460-Ex3 was more hydrophilic than HS-Em430-Ex2 and contained fewer alkyl chains but more acid groups. As a result, HS-Em460-Ex3 was eliminated more preferentially by NDMP anion exchange. However, compared with adsorption resins, the polarity of fluorescent components had a relatively small effect on the performance of anion exchange resin. The long-term pilot-scale experiments showed that the NDMP anion exchange process could remove approximately 30 % of the chemical oxygen demand and about 90 % of color from the biologically treated textile effluents. Once the issue of waste brine from resin desorption is solved, the NDMP anion exchange process could be a promising alternative for the advanced treatment of textile effluents.

Enhancement of fluorescence using nanoplasmonic and photonic structures

NASA Astrophysics Data System (ADS)

Arya, Akash; Tagore, Amit K.; Dantham, Venkata R.

2018-05-01

Two different nanoplasmonic structures have been synthesized using wet-chemistry method and demonstrated the enhancement of fluorescence of dye molecules. The difficulties associated with the plasmonic enhancement of fluorescence are discussed and to overcome these, an efficient approach has been proposed to enhance the fluorescence of a few molecules with the help of a high quality factor photonic microstructure (whispering gallery mode microresonator). The fabrication details of microresonators and experimental arrangement for enhancing the fluorescence are reported here.

Cavity-Enhanced Optical Readout of a Single Solid-State Spin

NASA Astrophysics Data System (ADS)

Sun, Shuo; Kim, Hyochul; Solomon, Glenn S.; Waks, Edo

2018-05-01

We demonstrate optical readout of a single spin using cavity quantum electrodynamics. The spin is based on a single trapped electron in a quantum dot that has a poor branching ratio of 0.43. Selectively coupling one of the optical transitions of the quantum dot to the cavity mode results in a spin-dependent cavity reflectivity that enables spin readout by monitoring the reflected intensity of an incident optical field. Using this approach, we demonstrate spin-readout fidelity of 0.61. Achieving this fidelity using resonance fluorescence from a bare dot would require 43 times improvement in photon collection efficiency.

Measurement of flow inside a vacuum cleaner head

NASA Astrophysics Data System (ADS)

Iguchi, Ryotaro; Ban, Hisataka; Sakakibara, Jun

2017-11-01

Vacuum cleaner head with rotating brushes is widely used as a home appliance. Although it efficiently collects dusts from the floor, flow field of the air and motion of the dust inside the head have not been fully investigated. In this study, we performed 3D-PIV (particle tracking velocimetry) measurement of velocity field inside the head. Water was used as working fluid, which allows a use of fluorescent particle to reduce unwanted reflection from the brushes and inner surface of the head. Mean velocity field and turbulence statistics in the head with and without the brush will be presented.

A simple but highly efficient multi-formyl phenol-amine system for fluorescence detection of peroxide explosive vapour.

PubMed

Xu, Wei; Fu, Yanyan; Gao, Yixun; Yao, Junjun; Fan, Tianchi; Zhu, Defeng; He, Qingguo; Cao, Huimin; Cheng, Jiangong

2015-07-11

A simple, highly stable, sensitive and selective fluorescent system for peroxide explosives was developed via an aromatic aldehyde oxidation reaction. The high efficiency arises from its higher HOMO level and multiple H-bonding. The sensitivity is obtained to be 0.1 ppt for H2O2 and 0.2 ppb for TATP.

Making Optical-Fiber Chemical Detectors More Sensitive

NASA Technical Reports Server (NTRS)

Rogowski, Robert S.; Egalon, Claudio O.

1993-01-01

Calculations based on exact theory of optical fiber shown how to increase optical efficiency and sensitivity of active-cladding step-index-profile optical-fiber fluorosensor using evanescent wave coupling. Optical-fiber fluorosensor contains molecules fluorescing when illuminated by suitable light in presence of analyte. Fluorescence coupled into and launched along core by evanescent-wave interaction. Efficiency increases with difference in refractive indices.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, A.N.; Benson, S.C.

1997-07-08

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figs.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, A.M.; Benson, S.C.

1998-06-16

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figs.

DNA complexes with dyes designed for energy transfer as fluorescent markers

DOEpatents

Glazer, A.N.; Benson, S.C.

1995-03-28

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figures.

Visualizing Epithelial Expression in Vertical and Horizontal Planes With Dual Axes Confocal Endomicroscope Using Compact Distal Scanner.

PubMed

Li, Gaoming; Li, Haijun; Duan, Xiyu; Zhou, Quan; Zhou, Juan; Oldham, Kenn R; Wang, Thomas D

2017-07-01

The epithelium is a thin layer of tissue that lines hollow organs, such as colon. Visualizing in vertical cross sections with sub-cellular resolution is essential to understanding early disease mechanisms that progress naturally in the plane perpendicular to the tissue surface. The dual axes confocal architecture collects optical sections in tissue by directing light at an angle incident to the surface using separate illumination and collection beams to reduce effects of scattering, enhance dynamic range, and increase imaging depth. This configuration allows for images to be collected in the vertical as well as horizontal planes. We designed a fast, compact monolithic scanner based on the principle of parametric resonance. The mirrors were fabricated using microelectromechanical systems (MEMS) technology and were coated with aluminum to maximize near-infrared reflectivity. We achieved large axial displacements [Formula: see text] and wide lateral deflections >20°. The MEMS chip has a 3.2×2.9 mm 2 form factor that allows for efficient packaging in the distal end of an endomicroscope. Imaging can be performed in either the vertical or horizontal planes with [Formula: see text] depth or 1 ×1 mm 2 area, respectively, at 5 frames/s. We systemically administered a Cy5.5-labeled peptide that is specific for EGFR, and collected near-infrared fluorescence images ex vivo from pre-malignant mouse colonic epithelium to reveal the spatial distribution of this molecular target. Here, we demonstrate a novel scanning mechanism in a dual axes confocal endomicroscope that collects optical sections of near-infrared fluorescence in either vertical or horizontal planes to visualize molecular expression in the epithelium.

PHYTOPLANKTON NUTRIENT STATUS AND VARIABLE FLUORESCENCE MEASUREMENTS IN A GULF COAST ESTUARY

EPA Science Inventory

Changes in variable fluorescence parameters such as the maximum quantum yield of fluorescence (a.k.a. photosynthetic efficiency and Fv/Fm) have been related to nutrient status in single-species cultures. To test if changes in Fv/Fm of mixed natural assemblages were related to nut...

Assessing FRET using Spectral Techniques

PubMed Central

Leavesley, Silas J.; Britain, Andrea L.; Cichon, Lauren K.; Nikolaev, Viacheslav O.; Rich, Thomas C.

2015-01-01

Förster resonance energy transfer (FRET) techniques have proven invaluable for probing the complex nature of protein–protein interactions, protein folding, and intracellular signaling events. These techniques have traditionally been implemented with the use of one or more fluorescence band-pass filters, either as fluorescence microscopy filter cubes, or as dichroic mirrors and band-pass filters in flow cytometry. In addition, new approaches for measuring FRET, such as fluorescence lifetime and acceptor photobleaching, have been developed. Hyperspectral techniques for imaging and flow cytometry have also shown to be promising for performing FRET measurements. In this study, we have compared traditional (filter-based) FRET approaches to three spectral-based approaches: the ratio of acceptor-to-donor peak emission, linear spectral unmixing, and linear spectral unmixing with a correction for direct acceptor excitation. All methods are estimates of FRET efficiency, except for one-filter set and three-filter set FRET indices, which are included for consistency with prior literature. In the first part of this study, spectrofluorimetric data were collected from a CFP–Epac–YFP FRET probe that has been used for intracellular cAMP measurements. All comparisons were performed using the same spectrofluorimetric datasets as input data, to provide a relevant comparison. Linear spectral unmixing resulted in measurements with the lowest coefficient of variation (0.10) as well as accurate fits using the Hill equation. FRET efficiency methods produced coefficients of variation of less than 0.20, while FRET indices produced coefficients of variation greater than 8.00. These results demonstrate that spectral FRET measurements provide improved response over standard, filter-based measurements. Using spectral approaches, single-cell measurements were conducted through hyperspectral confocal microscopy, linear unmixing, and cell segmentation with quantitative image analysis. Results from these studies confirmed that spectral imaging is effective for measuring subcellular, time-dependent FRET dynamics and that additional fluorescent signals can be readily separated from FRET signals, enabling multilabel studies of molecular interactions. PMID:23929684

Assessing FRET using spectral techniques.

PubMed

Leavesley, Silas J; Britain, Andrea L; Cichon, Lauren K; Nikolaev, Viacheslav O; Rich, Thomas C

2013-10-01

Förster resonance energy transfer (FRET) techniques have proven invaluable for probing the complex nature of protein-protein interactions, protein folding, and intracellular signaling events. These techniques have traditionally been implemented with the use of one or more fluorescence band-pass filters, either as fluorescence microscopy filter cubes, or as dichroic mirrors and band-pass filters in flow cytometry. In addition, new approaches for measuring FRET, such as fluorescence lifetime and acceptor photobleaching, have been developed. Hyperspectral techniques for imaging and flow cytometry have also shown to be promising for performing FRET measurements. In this study, we have compared traditional (filter-based) FRET approaches to three spectral-based approaches: the ratio of acceptor-to-donor peak emission, linear spectral unmixing, and linear spectral unmixing with a correction for direct acceptor excitation. All methods are estimates of FRET efficiency, except for one-filter set and three-filter set FRET indices, which are included for consistency with prior literature. In the first part of this study, spectrofluorimetric data were collected from a CFP-Epac-YFP FRET probe that has been used for intracellular cAMP measurements. All comparisons were performed using the same spectrofluorimetric datasets as input data, to provide a relevant comparison. Linear spectral unmixing resulted in measurements with the lowest coefficient of variation (0.10) as well as accurate fits using the Hill equation. FRET efficiency methods produced coefficients of variation of less than 0.20, while FRET indices produced coefficients of variation greater than 8.00. These results demonstrate that spectral FRET measurements provide improved response over standard, filter-based measurements. Using spectral approaches, single-cell measurements were conducted through hyperspectral confocal microscopy, linear unmixing, and cell segmentation with quantitative image analysis. Results from these studies confirmed that spectral imaging is effective for measuring subcellular, time-dependent FRET dynamics and that additional fluorescent signals can be readily separated from FRET signals, enabling multilabel studies of molecular interactions. © 2013 International Society for Advancement of Cytometry. Copyright © 2013 International Society for Advancement of Cytometry.

Analysis of Wilhelm Ostwald's "Colour Organ" with Raman microspectroscopy

NASA Astrophysics Data System (ADS)

Bridarolli, Alexandra; Atak, Sefkan; Herm, Christoph

2016-11-01

The "Scientific Colour Organ" is a collection of 680 pigment powders, created by the chemist Wilhelm Ostwald in 1925 as a means to represent his colour system. Today, it remains a leading part of colour theory. Analysis of these materials was undertaken to understand how the colour system was realised and to gain indications for preservation of the collection to which it belongs. Dispersive Raman microspectroscopy was applied directly to the powders, as well as using alternative techniques to suppress fluorescence. Barium sulphate was detected in all of the samples with one exception. Portable X-ray fluorescence revealed that this compound was a constituent of lithopone pigment. Raman spectroscopy furthermore revealed synthetic ultramarine (C.I. PB 29) as well as six different synthetic organic pigments and dyes (C.I. PY3; C.I. PO5; C.I. PR81:1; C.I. PV2 and two different triarylmethane dyes). Thin-layer chromatography was applied to determine the exact combination of dyes causing the gradual change in colour of each powder compared to the adjacent samples. With the exception of triarylmethane, the synthetic organic dyes could be identified with Raman spectroscopy directly on the chromatographic plate. The efficiency of thin-layer chromatography combined with Raman spectroscopy for identification of organic pigments could thus be shown. X-ray fluorescence indicated the presence of tungsten-molybdenum lakes in some samples. Comparison of the analytical results to information published by Oswald in 1917 showed that he switched to more light-stable synthetic organic pigments used for his "Scientific Colour Organ".

A wavelength-dispersive instrument for characterizing fluorescence and scattering spectra of individual aerosol particles on a substrate

NASA Astrophysics Data System (ADS)

Huffman, Donald R.; Swanson, Benjamin E.; Huffman, J. Alex

2016-08-01

We describe a novel, low-cost instrument to acquire both elastic and inelastic (fluorescent) scattering spectra from individual supermicron-size particles in a multi-particle collection on a microscope slide. The principle of the device is based on a slitless spectroscope that is often employed in astronomy to determine the spectra of individual stars in a star cluster but had not been applied to atmospheric particles. Under excitation, most commonly by either a 405 nm diode laser or a UV light-emitting diode (LED), fluorescence emission spectra of many individual particles can be determined simultaneously. The instrument can also acquire elastic scattering spectra from particles illuminated by a white-light source. The technique also provides the ability to detect and rapidly estimate the number fraction of fluorescent particles that could contaminate a collection of non-fluorescent material, even without analyzing full spectra. Advantages and disadvantages of using black-and-white cameras compared to color cameras are given. The primary motivation for this work has been to develop an inexpensive technique to characterize fluorescent biological aerosol particles, especially particles such as pollen and mold spores that can cause allergies. An example of an iPhone-enabled device is also shown as a means for collecting data on biological aerosols at lower cost or by utilizing citizen scientists for expanded data collection.

Radiative decay engineering 3. Surface plasmon-coupled directional emission

PubMed Central

Lakowicz, Joseph R.

2009-01-01

A new method of fluorescence detection that promises to increase sensitivity by 20- to 1000-fold is described. This method will also decrease the contribution of sample autofluorescence to the detected signal. The method depends on the coupling of excited fluorophores with the surface plasmon resonance present in thin metal films, typically silver and gold. The phenomenon of surface plasmon-coupled emission (SPCE) occurs for fluorophores 20–250 nm from the metal surface, allowing detection of fluorophores over substantial distances beyond the metal–sample interface. SPCE depends on interactions of the excited fluorophore with the metal surface. This interaction is independent of the mode of excitation; that is, it does not require evanescent wave or surface-plasmon excitation. In a sense, SPCE is the inverse process of the surface plasmon resonance absorption of thin metal films. Importantly, SPCE occurs over a narrow angular distribution, converting normally isotropic emission into easily collected directional emission. Up to 50% of the emission from unoriented samples can be collected, much larger than typical fluorescence collection efficiencies near 1% or less. SPCE is due only to fluorophores near the metal surface and may be regarded as emission from the induced surface plasmons. Autofluorescence from more distal parts of the sample is decreased due to decreased coupling. SPCE is highly polarized and autofluorescence can be further decreased by collecting only the polarized component or only the light propagating with the appropriate angle. Examples showing how simple optical configurations can be used in diagnostics, sensing, or biotechnology applications are presented. Surface plasmon-coupled emission is likely to find widespread applications throughout the biosciences. PMID:14690679

Efficient enzymatic synthesis and dual-colour fluorescent labelling of DNA probes using long chain azido-dUTP and BCN dyes

PubMed Central

Ren, Xiaomei; El-Sagheer, Afaf H.; Brown, Tom

2016-01-01

A sterically undemanding azide analogue of dTTP (AHP dUTP) with an alkyl chain and ethynyl attachment to the nucleobase was designed and incorporated into DNA by primer extension, reverse transcription and polymerase chain reaction (PCR). An azide-modified 523 bp PCR amplicon with all 335 thymidines replaced by AHP dU was shown to be a perfect copy of the template from which it was amplified. Replacement of thymidine with AHP dU increases duplex stability, accounting in part for the high incorporation efficiency of the azide-modified triphosphate. Single-stranded azide-labelled DNA was conveniently prepared from PCR products by λ-exonuclease digestion and streptavidin magnetic bead isolation. Efficient fluorescent labelling of single and double-stranded DNA was carried out using dyes functionalized with bicyclo[6.1.0]non-4-yne (BCN) via the strain-promoted alkyne-azide cycloaddition (SPAAC) reaction. This revealed that the degree of labelling must be carefully controlled to achieve optimum fluorescence and avoid fluorescence quenching. Dual-coloured probes were obtained in a single tube fluorescent labelling reaction; and varying the ratios of the two dyes provides a simple method to prepare DNA probes with unique fluorescent signatures. AHP dUTP is a versatile clickable nucleotide with potentially wide applications in biology and nanotechnology including single molecule studies and synthesis of modified aptamer libraries via SELEX. PMID:26819406

Novel insights into anoxic/aerobic(1)/aerobic(2) biological fluidized-bed system for coke wastewater treatment by fluorescence excitation-emission matrix spectra coupled with parallel factor analysis.

PubMed

Ou, Hua-Se; Wei, Chao-Hai; Mo, Ce-Hui; Wu, Hai-Zhen; Ren, Yuan; Feng, Chun-Hua

2014-10-01

Fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC) was applied to investigate the contaminant removal efficiency and fluorescent characteristic variations in a full scale coke wastewater (CWW) treatment plant with a novel anoxic/aerobic(1)/aerobic(2) (A/O(1)/O(2)) process, which combined with internal-loop fluidized-bed reactor. Routine monitoring results indicated that primary contaminants in CWW, such as phenols and free cyanide, were removed efficiently in A/O(1)/O(2) process (removal efficiency reached 99% and 95%, respectively). Three-dimensional excitation-emission matrix fluorescence spectroscopy and PARAFAC identified three fluorescent components, including two humic-like fluorescence components (C1 and C3) and one protein-like component (C2). Principal component analysis revealed that C1 and C2 correlated with COD (correlation coefficient (r)=0.782, p<0.01 and r=0.921, p<0.01), respectively) and phenols (r=0.796, p<0.01 and r=0.914, p<0.01, respectively), suggesting that C1 and C2 might be associated with the predominating aromatic contaminants in CWW. C3 correlated with mixed liquor suspended solids (r=0.863, p<0.01) in fluidized-bed reactors, suggesting that it might represent the biological dissolved organic matter. In A/O(1)/O(2) process, the fluorescence intensities of C1 and C2 consecutively decreased, indicating the degradation of aromatic contaminants. Correspondingly, the fluorescence intensity of C3 increased in aerobic(1) stage, suggesting an increase of biological dissolved organic matter. Copyright © 2014 Elsevier Ltd. All rights reserved.

Highly selective fluorescent and colorimetric chemosensor for detection of Hg2 + ion in aqueous media

NASA Astrophysics Data System (ADS)

Zareh Jonaghani, Mohammad; Zali-Boeini, Hassan

2017-05-01

A highly efficient and selective fluorescent and colorimetric chemosensor based on naphthothiazole skeleton was synthesized and its colorimetric and fluorescent properties were investigated. The sensor displays a rapid and highly selective colorimetric and fluorescence response toward Hg2 + without interference with other metal ions in CH3CN/H2O mixture (50/50, v/v). The detection limit for the fluorescent chemosensor S1 toward Hg2 + was 3.42 × 10- 8 M.

Antibody incubation at 37°C improves fluorescent immunolabeling in free-floating thick tissue sections.

PubMed

Xiao, Xia; Feng, Ya-Ping; Du, Bin; Sun, Han-Ru; Ding, You-Quan; Qi, Jian-Guo

2017-03-01

Fluorescent immunolabeling and imaging in free-floating thick (50-60 μm) tissue sections is relatively simple in practice and enables design-based non-biased stereology, or 3-D reconstruction and analysis. This method is widely used for 3-D in situ quantitative biology in many areas of biological research. However, the labeling quality and efficiency of standard protocols for fluorescent immunolabeling of these tissue sections are not always satisfactory. Here, we systematically evaluate the effects of raising the conventional antibody incubation temperatures (4°C or 21°C) to mammalian body temperature (37°C) in these protocols. Our modification significantly enhances the quality (labeling sensitivity, specificity, and homogeneity) and efficiency (antibody concentration and antibody incubation duration) of fluorescent immunolabeling of free-floating thick tissue sections.

Multicolor fluorescent biosensor for multiplexed detection of DNA.

PubMed

Hu, Rong; Liu, Tao; Zhang, Xiao-Bing; Huan, Shuang-Yan; Wu, Cuichen; Fu, Ting; Tan, Weihong

2014-05-20

Development of efficient methods for highly sensitive and rapid screening of specific oligonucleotide sequences is essential to the early diagnosis of serious diseases. In this work, an aggregated cationic perylene diimide (PDI) derivative was found to efficiently quench the fluorescence emission of a variety of anionic oligonucleotide-labeled fluorophores that emit at wavelengths from the visible to NIR region. This broad-spectrum quencher was then adopted to develop a multicolor biosensor via a label-free approach for multiplexed fluorescent detection of DNA. The aggregated perylene derivative exhibits a very high quenching efficiency on all ssDNA-labeled dyes associated with biosensor detection, having efficiency values of 98.3 ± 0.9%, 97 ± 1.1%, and 98.2 ± 0.6% for FAM, TAMRA, and Cy5, respectively. An exonuclease-assisted autocatalytic target recycling amplification was also integrated into the sensing system. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity toward target DNA, resulting in a detection limit of 20 pM, which is about 50-fold lower than that of traditional unamplified homogeneous fluorescent assay methods. The quencher did not interfere with the catalytic activity of nuclease, and the biosensor could be manipulated in either preaddition or postaddition manner with similar sensitivity. Moreover, the proposed sensing system allows for simultaneous and multicolor analysis of several oligonucleotides in homogeneous solution, demonstrating its potential application in the rapid screening of multiple biotargets.

[Laser Induced Fluorescence Spectroscopic Analysis of Aromatics from One Ring to Four Rings].

PubMed

Zhang, Peng; Liu, Hai-feng; Yue, Zong-yu; Chen, Bei-ling; Yao, Ming-fa

2015-06-01

In order to distinguish small aromatics preferably, a Nd : YAG Laser was used to supply an excitation laser, which was adjusted to 0.085 J x cm(-2) at 266 nm. Benzene, toluene, naphthalene, phenanthrene, anthracene, pyrene and chrysene were used as the representative of different rings aromatics. The fluorescence emission spectra were researched for each aromatic hydrocarbon and mixtures by Laser induced fluorescence (LIF). Results showed that the rings number determined the fluorescence emission spectra, and the structure with same rings number did not affect the emission fluorescence spectrum ranges. This was due to the fact that the absorption efficiency difference at 266 nm resulted in that the fluorescence intensities of each aromatic hydrocarbon with same rings number were different and the fluorescence intensities difference were more apparently with aromatic ring number increasing. When the absorption efficiency was similar at 266 nm and the concentrations of each aromatic hydrocarbon were same, the fluorescence intensities were increased with aromatic ring number increasing. With aromatic ring number increasing, the fluorescence spectrum and emission peak wavelength were all red-shifted from ultraviolet to visible and the fluorescence spectrum range was also wider as the absorption efficiency was similar. The fluorescence emission spectra from one to four rings could be discriminated in the following wavelengths, 275 to 320 nm, 320 to 375 nm, 375 to 425 nm, 425 to 556 nm, respectively. It can be used for distinguish the type of the polycyclic aromatic hydrocarbons (PAHs) as it exists in single type. As PAHs are usually exist in a variety of different rings number at the same time, the results for each aromatic hydrocarbon may not apply to the aromatic hydrocarbon mixtures. For the aromatic hydrocarbon mixtures, results showed that the one- or two-ring PAHs in mixtures could not be detected by fluorescence as three- or four-ring PAHs existed in mixture. This was caused by radiation energy transfer mechanism, in which the ultraviolet light was lost in mixtures but the fluorescence intensities were increased with the one- or two-ring PAHs adding. When the mixture only contained three- and four-ring PAHs, the fluorescence emission spectrum showed the both characteristics of three- and four-ring PAHs fluorescence. When three- and four-ring PAHs existed in mixtures at the same time, the fluorescence emission spectra were related to each concentration, so the rings number could be discriminated to a certain extent.

New Frontiers in NanoBiotechnology: Monitoring the Protein Function With Single Protein Resolution

DTIC Science & Technology

2005-03-29

Protein (GFP) is a spontaneously fluorescent polypeptide of 27 kD from the jellyfish Aequorea victoria that absorbs UV-blue light and emits in the...will have vast applications in science. Relationship between structure and optical properties in Green Fluorescent Proteins : A quantum mechanical study...RESEARCH AND DEVELOPMENT Invited talks Folding, stability and fluorescence efficiency of the Green and Red Fluorescent Proteins Saverio Alberti Lab.

Thermally activated delayed fluorescence of fluorescein derivative for time-resolved and confocal fluorescence imaging.

PubMed

Xiong, Xiaoqing; Song, Fengling; Wang, Jingyun; Zhang, Yukang; Xue, Yingying; Sun, Liangliang; Jiang, Na; Gao, Pan; Tian, Lu; Peng, Xiaojun

2014-07-09

Compared with fluorescence imaging utilizing fluorophores whose lifetimes are in the order of nanoseconds, time-resolved fluorescence microscopy has more advantages in monitoring target fluorescence. In this work, compound DCF-MPYM, which is based on a fluorescein derivative, showed long-lived luminescence (22.11 μs in deaerated ethanol) and was used in time-resolved fluorescence imaging in living cells. Both nanosecond time-resolved transient difference absorption spectra and time-correlated single-photon counting (TCSPC) were employed to explain the long lifetime of the compound, which is rare in pure organic fluorophores without rare earth metals and heavy atoms. A mechanism of thermally activated delayed fluorescence (TADF) that considers the long wavelength fluorescence, large Stokes shift, and long-lived triplet state of DCF-MPYM was proposed. The energy gap (ΔEST) of DCF-MPYM between the singlet and triplet state was determined to be 28.36 meV by the decay rate of DF as a function of temperature. The ΔE(ST) was small enough to allow efficient intersystem crossing (ISC) and reverse ISC, leading to efficient TADF at room temperature. The straightforward synthesis of DCF-MPYM and wide availability of its starting materials contribute to the excellent potential of the compound to replace luminescent lanthanide complexes in future time-resolved imaging technologies.

Study of improving signal-noise ratio for fluorescence channel

NASA Astrophysics Data System (ADS)

Wang, Guoqing; Li, Xin; Lou, Yue; Chen, Dong; Zhao, Xin; Wang, Ran; Yan, Debao; Zhao, Qi

2017-10-01

Laser-induced fluorescence(LIFS), which is one of most effective discrimination methods to identify the material at the molecular level by inducing fluorescence spectrum, has been popularized for its fast and accurate probe's results. According to the research, violet laser or ultraviolet laser is always used as excitation light source. While, There is no atmospheric window for violet laser and ultraviolet laser, causing laser attenuation along its propagation path. What's worse, as the laser reaching sample, part of the light is reflected. That is, excitation laser really react on sample to produce fluorescence is very poor, leading to weak fluorescence mingled with the background light collected by LIFS' processing unit, when it used outdoor. In order to spread LIFS to remote probing under the complex background, study of improving signal-noise ratio for fluorescence channel is a meaningful work. Enhancing the fluorescence intensity and inhibiting background light both can improve fluorescence' signal-noise ratio. In this article, three different approaches of inhibiting background light are discussed to improve the signal-noise ratio of LIFS. The first method is increasing fluorescence excitation area in the proportion of LIFS' collecting field by expanding laser beam, if the collecting filed is fixed. The second one is changing field angle base to accommodate laser divergence angle. The third one is setting a very narrow gating circuit to control acquisition circuit, which is shortly open only when fluorescence arriving. At some level, these methods all can reduce the background light. But after discussion, the third one is best with adding gating acquisition circuit to acquisition circuit instead of changing light path, which is effective and economic.

Efficient and Rapid Isolation of Early-stage Embryos from Arabidopsis thaliana Seeds

PubMed Central

Raissig, Michael T.; Gagliardini, Valeria; Jaenisch, Johan; Grossniklaus, Ueli; Baroux, Célia

2013-01-01

In flowering plants, the embryo develops within a nourishing tissue - the endosperm - surrounded by the maternal seed integuments (or seed coat). As a consequence, the isolation of plant embryos at early stages (1 cell to globular stage) is technically challenging due to their relative inaccessibility. Efficient manual dissection at early stages is strongly impaired by the small size of young Arabidopsis seeds and the adhesiveness of the embryo to the surrounding tissues. Here, we describe a method that allows the efficient isolation of young Arabidopsis embryos, yielding up to 40 embryos in 1 hr to 4 hr, depending on the downstream application. Embryos are released into isolation buffer by slightly crushing 250-750 seeds with a plastic pestle in an Eppendorf tube. A glass microcapillary attached to either a standard laboratory pipette (via a rubber tube) or a hydraulically controlled microinjector is used to collect embryos from droplets placed on a multi-well slide on an inverted light microscope. The technical skills required are simple and easily transferable, and the basic setup does not require costly equipment. Collected embryos are suitable for a variety of downstream applications such as RT-PCR, RNA sequencing, DNA methylation analyses, fluorescence in situ hybridization (FISH), immunostaining, and reporter gene assays. PMID:23770918

Efficiency of the intermolecular interaction of salicylic acid neutral form and monoanion with Cd2 + ion studied by methods of absorption and fluorescence

NASA Astrophysics Data System (ADS)

Lavrik, N. L.; Mulloev, N. U.

2018-02-01

The methods of absorption and fluorescence were used to study the efficiency of the interaction between salicylic acid derivatives SAD (neutral SA form and SA monoanion) and Cd2 + ions (in CdBr2 salt) within the range pH = 1.5 ÷ 8. The efficiency was determined from the change in both the absorption band contour and the fluorescence intensity of various SAD forms. It has been established that depending on the SAD form, the addition of CdBr2 to a starting solution leads to the formation of additional absorption for both the shorter wave lengths in the absorption spectrum of the neutral form (at pH < 3) and the longer wave lengths in the absorption spectrum for the HSal- monoanion (at pH > 4). In the fluorescence spectra, the intensity was observed to increase for the neutral SAD form (at pH < 3) and to decrease for the HSal- monoanion (at pH > 4) after addition of CdBr2. The spectral changes were interpreted in the framework of common notions about the effect of three physicochemical factors that determine the interaction between the SAD and the Cd2 + ion and affect the parameters of absorption and fluorescence spectra. These factors are: (1) the decrease in pH after addition of CdBr2 to the SAD solution, (2) the decrease in the efficiency of the H-bonding of SAD molecules to the water ones, and (3) the existence of electrostatic ion-ion interaction between the HSal- monoanion and the Cd2 + ion. The bimolecular fluorescence quenching constants Kq of HSal- monoanion fluorescence quenching by the Cd2 + ion appeared to be substantially less than those of the quenching which would follow either the dynamic (diffusion) or the concentration (static) mechanisms.

Injectant mole-fraction imaging in compressible mixing flows using planar laser-induced iodine fluorescence

NASA Technical Reports Server (NTRS)

Hartfield, Roy J., Jr.; Abbitt, John D., III; Mcdaniel, James C.

1989-01-01

A technique is described for imaging the injectant mole-fraction distribution in nonreacting compressible mixing flow fields. Planar fluorescence from iodine, seeded into air, is induced by a broadband argon-ion laser and collected using an intensified charge-injection-device array camera. The technique eliminates the thermodynamic dependence of the iodine fluorescence in the compressible flow field by taking the ratio of two images collected with identical thermodynamic flow conditions but different iodine seeding conditions.

Micro-Tom Tomato as an Alternative Plant Model System: Mutant Collection and Efficient Transformation.

PubMed

Shikata, Masahito; Ezura, Hiroshi

2016-01-01

Tomato is a model plant for fruit development, a unique feature that classical model plants such as Arabidopsis and rice do not have. The tomato genome was sequenced in 2012 and tomato is becoming very popular as an alternative system for plant research. Among many varieties of tomato, Micro-Tom has been recognized as a model cultivar for tomato research because it shares some key advantages with Arabidopsis including its small size, short life cycle, and capacity to grow under fluorescent lights at a high density. Mutants and transgenic plants are essential materials for functional genomics research, and therefore, the availability of mutant resources and methods for genetic transformation are key tools to facilitate tomato research. Here, we introduce the Micro-Tom mutant database "TOMATOMA" and an efficient transformation protocol for Micro-Tom.

Quantification of free cysteines in membrane and soluble proteins using a fluorescent dye and thermal unfolding.

PubMed

Branigan, Emma; Pliotas, Christos; Hagelueken, Gregor; Naismith, James H

2013-11-01

Cysteine is an extremely useful site for selective attachment of labels to proteins for many applications, including the study of protein structure in solution by electron paramagnetic resonance (EPR), fluorescence spectroscopy and medical imaging. The demand for quantitative data for these applications means that it is important to determine the extent of the cysteine labeling. The efficiency of labeling is sensitive to the 3D context of cysteine within the protein. Where the label or modification is not directly measurable by optical or magnetic spectroscopy, for example, in cysteine modification to dehydroalanine, assessing labeling efficiency is difficult. We describe a simple assay for determining the efficiency of modification of cysteine residues, which is based on an approach previously used to determine membrane protein stability. The assay involves a reaction between the thermally unfolded protein and a thiol-specific coumarin fluorophore that is only fluorescent upon conjugation with thiols. Monitoring fluorescence during thermal denaturation of the protein in the presence of the dye identifies the temperature at which the maximum fluorescence occurs; this temperature differs among proteins. Comparison of the fluorescence intensity at the identified temperature between modified, unmodified (positive control) and cysteine-less protein (negative control) allows for the quantification of free cysteine. We have quantified both site-directed spin labeling and dehydroalanine formation. The method relies on a commonly available fluorescence 96-well plate reader, which rapidly screens numerous samples within 1.5 h and uses <100 μg of material. The approach is robust for both soluble and detergent-solubilized membrane proteins.

Development of a dose-controlled multiculture cell exposure chamber for efficient delivery of airborne and engineered nanoparticles

NASA Astrophysics Data System (ADS)

Asimakopoulou, Akrivi; Daskalos, Emmanouil; Lewinski, Nastassja; Riediker, Michael; Papaioannou, Eleni; Konstandopoulos, Athanasios G.

2013-04-01

In order to study the various health influencing parameters related to engineered nanoparticles as well as to soot emitted by Diesel engines, there is an urgent need for appropriate sampling devices and methods for cell exposure studies that simulate the respiratory system and facilitate associated biological and toxicological tests. The objective of the present work was the further advancement of a Multiculture Exposure Chamber (MEC) into a dose-controlled system for efficient delivery of nanoparticles to cells. It was validated with various types of nanoparticles (Diesel engine soot aggregates, engineered nanoparticles for various applications) and with state-of-the-art nanoparticle measurement instrumentation to assess the local deposition of nanoparticles on the cell cultures. The dose of nanoparticles to which cell cultures are being exposed was evaluated in the normal operation of the in vitro cell culture exposure chamber based on measurements of the size specific nanoparticle collection efficiency of a cell free device. The average efficiency in delivering nanoparticles in the MEC was approximately 82%. The nanoparticle deposition was demonstrated by Transmission Electron Microscopy (TEM). Analysis and design of the MEC employs Computational Fluid Dynamics (CFD) and true to geometry representations of nanoparticles with the aim to assess the uniformity of nanoparticle deposition among the culture wells. Final testing of the dose-controlled cell exposure system was performed by exposing A549 lung cell cultures to fluorescently labeled nanoparticles. Delivery of aerosolized nanoparticles was demonstrated by visualization of the nanoparticle fluorescence in the cell cultures following exposure. Also monitored was the potential of the aerosolized nanoparticles to generate reactive oxygen species (ROS) (e.g. free radicals and peroxides generation), thus expressing the oxidative stress of the cells which can cause extensive cellular damage or damage on DNA.

Detection of munitions grade g-series nerve agents using Raman excitation at 1064 nm

NASA Astrophysics Data System (ADS)

Roy, Eric; Wilcox, Phillip G.; Hoffland, Soren; Pardoe, Ian

2015-05-01

Raman spectroscopy is a powerful tool for obtaining molecular structure information of a sample. While Raman spectroscopy is a common laboratory based analytical tool, miniaturization of opto-electronic components has allowed handheld Raman analyzers to become commercially available. These handheld systems are utilized by Military and First Responder operators tasked with rapidly identifying potentially hazardous chemicals in the field. However, one limitation of many handheld Raman detection systems is strong interference caused by fluorescence of the sample or underlying surface which obscures the characteristic Raman signature of the target analyte. Munitions grade chemical warfare agents (CWAs) are produced and stored in large batches and typically have more impurities from the storage container, degradation, or unreacted precursors. In this work, Raman spectra of munitions grade CWAs were collected using a handheld Raman spectrometer with a 1064 nm excitation laser. While Raman scattering generated by a 1064 nm laser is inherently less efficient than excitation at shorter wavelengths, high quality spectra were easily obtained due to significantly reduced fluorescence of the munitions grade CWAs. The spectra of these less pure, but more operationally relevant, munitions grade CWAs were then compared to spectra of CASARM grade CWAs, as well as Raman spectra collected using the more common 785 nm excitation laser.

Photodynamic effects on human and chicken erythrocytes

NASA Astrophysics Data System (ADS)

Kimel, Sol; Koenig, Karsten; Berns, Michael W.

1995-02-01

The intracellular accumulation of a variety of photosensitizers in human (non-nucleated) and chicken (nucleated) erythrocytes, as well as the photodynamically induced hemolysis were studied using 488 nm laser microirradiation (15 (mu) W, 100X) and confocal laser scanning fluorescence microscopy. Cells incubated with the negatively charged hydrophilic compounds TPPS4 and Pd-TPPS4 exhibited no significant fluorescence before irradiation, but developed strong fluorescence in the cellular and nuclear membranes following photoinduced membrane damage. In contrast, microirradiation of Photofrin-incubated erythrocytes showed instantaneous fluorescence which decreased due to photodegradation. For the cationic, hydrophilic dye Methylene Blue, significant fluorescence was detected in the nucleus only. Following ALA incubation, large intercellular differences were observed in fluorescence in the red spectral region. These differences are probably due to the differential ability of individual erythrocytes to biosynthesize protoporphyrin IX. Photofrin was the most efficient photosensitizer to induce hemolysis. Higher radiant exposures were required for lysis of nucleated than of human red blood cells, except in the case of Methylene Blue. Irradiation was more efficient for unwashed cell suspensions than for washed suspensions, indicating the non-negligible role of extracellular photosensitizing molecules.

pH-Responsive Fluorescence Enhancement in Graphene Oxide-Naphthalimide Nanoconjugates: A Fluorescence Turn-On Sensor for Acetylcholine.

PubMed

Mangalath, Sreejith; Abraham, Silja; Joseph, Joshy

2017-08-22

A pH-sensitive, fluorescence "turn-on" sensor based on a graphene oxide-naphthalimide (GO-NI) nanoconjugate for the detection of acetylcholine (ACh) by monitoring the enzymatic activity of acetylcholinesterase (AChE) in aqueous solution is reported. These nanoconjugates were synthesized by covalently anchoring picolyl-substituted NI derivatives on the GO/reduced GO surface through a 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide coupling strategy, and the morphological and photophysical properties were studied in detail. Synergistic effects of π-π interactions between GO and the NI chromophore, and efficient photoinduced electron- and energy-transfer processes, were responsible for the strong quenching of fluorescence of these nanoconjugates, which were perturbed under acidic pH conditions, leading to significant enhancement of fluorescence emission. This nanoconjugate was successfully employed for the efficient sensing of pH changes caused by the enzymatic activity of AChE, thereby demonstrating its utility as a fluorescence turn-on sensor for ACh in the neurophysiological range. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Evaluation of the charge transfer efficiency of organic thin-film photovoltaic devices fabricated using a photoprecursor approach.

PubMed

Masuo, Sadahiro; Sato, Wataru; Yamaguchi, Yuji; Suzuki, Mitsuharu; Nakayama, Ken-ichi; Yamada, Hiroko

2015-05-01

Recently, a unique 'photoprecursor approach' was reported as a new option to fabricate a p-i-n triple-layer organic photovoltaic device (OPV) through solution processes. By fabricating the p-i-n architecture using two kinds of photoprecursors and a [6,6]-phenyl C71 butyric acid methyl ester (PC71BM) as the donor and the acceptor, the p-i-n OPVs afforded a higher photovoltaic efficiency than the corresponding p-n devices and i-devices, while the photovoltaic efficiency of p-i-n OPVs depended on the photoprecursors. In this work, the charge transfer efficiency of the i-devices composed of the photoprecursors and PC71BM was investigated using high-sensitivity fluorescence microspectroscopy combined with a time-correlated single photon counting technique to elucidate the photovoltaic efficiency depending on the photoprecursors and the effects of the p-i-n architecture. The spatially resolved fluorescence images and fluorescence lifetime measurements clearly indicated that the compatibility of the photoprecursors with PC71BM influences the charge transfer and the photovoltaic efficiencies. Although the charge transfer efficiency of the i-device was quite high, the photovoltaic efficiency of the i-device was much lower than that of the p-i-n device. These results imply that the carrier generation and carrier transportation efficiencies can be increased by fabricating the p-i-n architecture.

Green-emitting MADF complex for OLED applications

NASA Astrophysics Data System (ADS)

Klimes, Kody; Zhu, Zhi-Qiang; Holloway, Sean; Li, Jian

2016-09-01

In this article, we demonstrated an exceptional palladium complex that exhibits both phosphorescence and delayed fluorescence for use as an efficient emitter in OLEDs. Devices employing PdN3N achieved external quantum efficiencies in excess of 22% and remarkable device operational lifetime to 90% initial luminance estimated at over 30,000 h at a practical luminance of 100 cd/m2. Further tuning of the phosphorescent and delayed fluorescent emission should have a great impact in the development of efficient and stable emitters for deep blue or white OLEDs.

Nanoscale Plasmonic V-Groove Waveguides for the Interrogation of Single Fluorescent Bacterial Cells.

PubMed

Lotan, Oren; Bar-David, Jonathan; Smith, Cameron L C; Yagur-Kroll, Sharon; Belkin, Shimshon; Kristensen, Anders; Levy, Uriel

2017-09-13

We experimentally demonstrate the interrogation of an individual Escherichia coli cell using a nanoscale plasmonic V-groove waveguide. Several different configurations were studied. The first involved the excitation of the cell in a liquid environment because it flows on top of the waveguide nanocoupler, while the obtained fluorescence is coupled into the waveguide and collected at the other nanocoupler. The other two configurations involved the positioning of the bacterium within the nanoscale waveguide and its excitation in a dry environment either directly from the top or through waveguide modes. This is achieved by taking advantage of the waveguide properties not only for light guiding but also as a mechanical tool for trapping the bacteria within the V-grooves. The obtained results are supported by a set of numerical simulations, shedding more light on the mechanism of excitation. This demonstration paves the way for the construction of an efficient bioplasmonic chip for diverse cell-based sensing applications.

Fluorescent Amplified-Fragment Length Polymorphism Genotyping of Neisseria meningitidis Identifies Clones Associated with Invasive Disease

PubMed Central

Goulding, Jonathan N.; Hookey, John V.; Stanley, John; Olver, Will; Neal, Keith R.; Ala'Aldeen, Dlawer A. A.; Arnold, Catherine

2000-01-01

Fluorescent amplified-fragment length polymorphism (FAFLP), a genotyping technique with phylogenetic significance, was applied to 123 isolates of Neisseria meningitidis. Nine of these were from an outbreak in a British university; 9 were from a recent outbreak in Pontypridd, Glamorgan; 15 were from sporadic cases of meningococcal disease; 26 were from the National Collection of Type Cultures; 58 were carrier isolates from Ironville, Derbyshire; 1 was a disease isolate from Ironville; and five were representatives of invasive clones of N. meningitidis. FAFLP analysis results were compared with previously published multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) results. FAFLP was able to identify hypervirulent, hyperendemic lineages (invasive clones) of N. meningitidis as well as did MLST. PFGE did not discriminate between two strains from the outbreak that were classified as similar but distinct by FAFLP. The results suggest that high resolution of N. meningitidis for outbreak and other epidemiological analyses is more cost efficient by FAFLP than by sequencing procedures. PMID:11101599

White organic light-emitting diodes with fluorescent tube efficiency.

PubMed

Reineke, Sebastian; Lindner, Frank; Schwartz, Gregor; Seidler, Nico; Walzer, Karsten; Lüssem, Björn; Leo, Karl

2009-05-14

The development of white organic light-emitting diodes (OLEDs) holds great promise for the production of highly efficient large-area light sources. High internal quantum efficiencies for the conversion of electrical energy to light have been realized. Nevertheless, the overall device power efficiencies are still considerably below the 60-70 lumens per watt of fluorescent tubes, which is the current benchmark for novel light sources. Although some reports about highly power-efficient white OLEDs exist, details about structure and the measurement conditions of these structures have not been fully disclosed: the highest power efficiency reported in the scientific literature is 44 lm W(-1) (ref. 7). Here we report an improved OLED structure which reaches fluorescent tube efficiency. By combining a carefully chosen emitter layer with high-refractive-index substrates, and using a periodic outcoupling structure, we achieve a device power efficiency of 90 lm W(-1) at 1,000 candelas per square metre. This efficiency has the potential to be raised to 124 lm W(-1) if the light outcoupling can be further improved. Besides approaching internal quantum efficiency values of one, we have also focused on reducing energetic and ohmic losses that occur during electron-photon conversion. We anticipate that our results will be a starting point for further research, leading to white OLEDs having efficiencies beyond 100 lm W(-1). This could make white-light OLEDs, with their soft area light and high colour-rendering qualities, the light sources of choice for the future.

High-efficiency electroluminescence and amplified spontaneous emission from a thermally activated delayed fluorescent near-infrared emitter

NASA Astrophysics Data System (ADS)

Kim, Dae-Hyeon; D'Aléo, Anthony; Chen, Xian-Kai; Sandanayaka, Atula D. S.; Yao, Dandan; Zhao, Li; Komino, Takeshi; Zaborova, Elena; Canard, Gabriel; Tsuchiya, Youichi; Choi, Eunyoung; Wu, Jeong Weon; Fages, Frédéric; Brédas, Jean-Luc; Ribierre, Jean-Charles; Adachi, Chihaya

2018-02-01

Near-infrared organic light-emitting diodes and semiconductor lasers could benefit a variety of applications including night-vision displays, sensors and information-secured displays. Organic dyes can generate electroluminescence efficiently at visible wavelengths, but organic light-emitting diodes are still underperforming in the near-infrared region. Here, we report thermally activated delayed fluorescent organic light-emitting diodes that operate at near-infrared wavelengths with a maximum external quantum efficiency of nearly 10% using a boron difluoride curcuminoid derivative. As well as an effective upconversion from triplet to singlet excited states due to the non-adiabatic coupling effect, this donor-acceptor-donor compound also exhibits efficient amplified spontaneous emission. By controlling the polarity of the active medium, the maximum emission wavelength of the electroluminescence spectrum can be tuned from 700 to 780 nm. This study represents an important advance in near-infrared organic light-emitting diodes and the design of alternative molecular architectures for photonic applications based on thermally activated delayed fluorescence.

Advanced Imaging Approaches to Characterize Stromal and Metabolic Changes in In Vivo Mammary Tumor Models

DTIC Science & Technology

2013-03-01

characterization and toward future intravital studies. Preliminary fluorescence lifetime images were also collected intravitally through a mammary imaging window...intend to use this characterization to understand shifts in fluorescence lifetime collected by intravital imaging using a mammary imaging window...collected intravitally through a mammary imaging window implanted in a female, PyVT positive, Col1a1 heterozygote, mouse (Figure 7). A paper has

The Non-Specific Binding of Fluorescent-Labeled MiRNAs on Cell Surface by Hydrophobic Interaction.

PubMed

Lu, Ting; Lin, Zongwei; Ren, Jianwei; Yao, Peng; Wang, Xiaowei; Wang, Zhe; Zhang, Qunye

2016-01-01

MicroRNAs are small noncoding RNAs about 22 nt long that play key roles in almost all biological processes and diseases. The fluorescent labeling and lipofection are two common methods for changing the levels and locating the position of cellular miRNAs. Despite many studies about the mechanism of DNA/RNA lipofection, little is known about the characteristics, mechanisms and specificity of lipofection of fluorescent-labeled miRNAs. Therefore, miRNAs labeled with different fluorescent dyes were transfected into adherent and suspension cells using lipofection reagent. Then, the non-specific binding and its mechanism were investigated by flow cytometer and laser confocal microscopy. The results showed that miRNAs labeled with Cy5 (cyanine fluorescent dye) could firmly bind to the surface of adherent cells (Hela) and suspended cells (K562) even without lipofection reagent. The binding of miRNAs labeled with FAM (carboxyl fluorescein) to K562 cells was obvious, but it was not significant in Hela cells. After lipofectamine reagent was added, most of the fluorescently labeled miRNAs binding to the surface of Hela cells were transfected into intra-cell because of the high transfection efficiency, however, most of them were still binding to the surface of K562 cells. Moreover, the high-salt buffer which could destroy the electrostatic interactions did not affect the above-mentioned non-specific binding, but the organic solvent which could destroy the hydrophobic interactions eliminated it. These results implied that the fluorescent-labeled miRNAs could non-specifically bind to the cell surface by hydrophobic interaction. It would lead to significant errors in the estimation of transfection efficiency only according to the cellular fluorescence intensity. Therefore, other methods to evaluate the transfection efficiency and more appropriate fluorescent dyes should be used according to the cell types for the accuracy of results.

Use of a multiseparation fiber optic probe for the optical diagnosis of breast cancer.

PubMed

Zhu, Changfang; Palmer, Gregory M; Breslin, Tara M; Xu, Fushen; Ramanujam, Nirmala

2005-01-01

We explore the effects of the illumination and collection geometry on optical spectroscopic diagnosis of breast cancer. Fluorescence and diffuse reflectance spectroscopy in the UV-visible spectral range are made with a multiseparation probe at three illumination-collection separations of 735, 980, and 1225 microm, respectively, from 13 malignant and 34 nonmalignant breast tissues. Statistical analysis is carried out on two types of data inputs: (1) the fluorescence and diffuse reflectance spectra recorded at each of the three illumination-collection separations and (2) the integrated fluorescence (at each excitation wavelength) or diffuse reflectance over the entire spectrum at all three illumination-collection separations. The results show that using the integrated fluorescence intensities recorded at a single excitation wavelength at all three illumination-collection separations can discriminate malignant from nonmalignant breast tissues with similar classification accuracy to that using spectral data measured at several excitation wavelengths with a single illumination-collection separation. These findings have significant implications with respect to the design of an optical system for breast cancer diagnosis. Examining the intensity attenuation at a single wavelength rather than spectral intensities at multiple wavelengths can significantly reduce the measurement and data processing time in a clinical setting as well as the cost and complexity of the optical system. Copyright 2005 Society of Photo-Optical Instrumentation Engineers.

Efficient HOMO-LUMO separation by multiple resonance effect toward ultrapure blue thermally activated delayed fluorescence

NASA Astrophysics Data System (ADS)

Hatakeyama, Takuji; Ikuta, Toshiaki; Shiren, Kazushi; Nakajima, Kiichi; Nomura, Shintaro; Ni, Jingping

2016-09-01

Organic light-emitting diodes (OLEDs) play an important role in the new generation of flat-panel displays. Conventional OLEDs employing fluorescent materials together with triplet-triplet annihilation suffer from a relatively low internal quantum efficiency (IQE) of 62.5%. On the other hand, the IQE of OLEDs employing phosphorescent or thermally activated delayed fluorescence (TADF) materials can reach 100%. However, these materials exhibit very broad peaks with a full-width at half-maximum (FWHM) of 70-100 nm and cannot satisfy the color-purity requirements for displays. Therefore, the latest commercial OLED displays employ blue fluorescent materials with a relatively low IQE, and efficient blue emitters with a small FWHM are highly needed. In our manuscript, we present organic molecules that exhibit ultrapure blue fluorescence based on TADF. These molecules consist of three benzene rings connected by one boron and two nitrogen atoms, which establish a rigid polycyclic framework and significant localization of the highest occupied and lowest unoccupied molecular orbitals by a multiple resonance effect. An OLED device based on the new emitter exhibits ultrapure blue emission at 467 nm with an FWHM of 28 nm, Commission Internationale de l'Eclairage (CIE) coordinates of (0.12, 0.13), and an IQE of 100%, which represent record-setting performance for blue OLED devices.

Immunocytochemistry and fluorescence imaging efficiently identify individual neurons with CRISPR/Cas9-mediated gene disruption in primary cortical cultures.

PubMed

Tsunematsu, Hiroto; Uyeda, Akiko; Yamamoto, Nobuhiko; Sugo, Noriyuki

2017-08-01

CRISPR/Cas9 system is a powerful method to investigate the role of genes by introducing a mutation selectively and efficiently to specific genome positions in cell and animal lines. However, in primary neuron cultures, this method is affected by the issue that the effectiveness of CRISPR/Cas9 is different in each neuron. Here, we report an easy, quick and reliable method to identify mutants induced by the CRISPR/Cas9 system at a single neuron level, using immunocytochemistry (ICC) and fluorescence imaging. Dissociated cortical cells were transfected with CRISPR/Cas9 plasmids targeting the transcription factor cAMP-response element binding protein (CREB). Fluorescence ICC with CREB antibody and quantitative analysis of fluorescence intensity demonstrated that CREB expression disappeared in a fraction of the transfected neurons. The downstream FOS expression was also decreased in accordance with suppressed CREB expression. Moreover, dendritic arborization was decreased in the transfected neurons which lacked CREB immunoreactivity. Detection of protein expression is efficient to identify individual postmitotic neurons with CRISPR/Cas9-mediated gene disruption in primary cortical cultures. The present method composed of CRISPR/Cas9 system, ICC and fluorescence imaging is applicable to study the function of various genes at a single-neuron level.

Transfer of ultraviolet photon energy into fluorescent light in the visible path represents a new and efficient protection mechanism of sunscreens

NASA Astrophysics Data System (ADS)

Vergou, Theognosia; Patzelt, Alexa; Richter, Heike; Schanzer, Sabine; Zastrow, Leonhard; Golz, Karin; Doucet, Olivier; Antoniou, Christina; Sterry, Wolfram; Lademann, Juergen

2011-10-01

The development of sunscreens with high sun protection factor (SPF) values but low filter concentrations is the ultimate goal. The purpose of the present study was to investigate why a sunscreen spray and cream with different concentrations of the same UV-filters provided the same SPF. Therefore, the homogeneity of the distribution of both sunscreens was investigated by laser scanning microscopy (LSM) and tape stripping (TS). Additionally, the energy transfer mechanisms of the sunscreens on the skin were analyzed. The TS and LSM showed a better homogeneity of the distribution of the spray. With Wood's light, a total absorption of the irradiation was detected in the spray area. In contrast, after cream treatment, an intensive fluorescent signal was observed. It was demonstrated that this fluorescent signal was caused by nonthermal energy transferred from the UV-filters to one compound of the cream releasing its excitation energy by fluorescence. This nonthermal energy transfer seemed to be the reason for the high efficiency of the cream, which is subjected to thermal relaxation. The transfer of UV photon energy into fluorescent light represents a new approach to increase the efficiency of sunscreens and could form the basis for a new generation of sunscreens.

The efficiency of non-photochemical fluorescence quenching by cation radicals in photosystem II reaction centers.

PubMed

Paschenko, V Z; Churin, A A; Gorokhov, V V; Grishanova, N P; Korvatovskii, B N; Maksimov, E G; Mamedov, M D

2016-12-01

In a direct experiment, the rate constants of photochemical k p and non-photochemical k p + quenching of the chlorophyll fluorescence have been determined in spinach photosystem II (PS II) membrane fragments, oxygen-evolving PS II core, as well as manganese-depleted PS II particles using pulse fluorimetry. In the dark-adapted reaction center(s) (RC), the fluorescence decay kinetics of the antenna were measured at low-intensity picosecond pulsed excitation. To create a "closed" P680 + Q A - state, RCs were illuminated by high-intensity actinic flash 8 ns prior to the measuring flash. The obtained data were approximated by the sum of two decaying exponents. It was found that the antennae fluorescence quenching efficiency by the oxidized photoactive pigment of RC P680 + was about 1.5 times higher than that of the neutral P680 state. These results were confirmed by a single-photon counting technique, which allowed to resolve the additional slow component of the fluorescence decay. Slow component was assigned to the charge recombination of P680 + Pheo - in PS II RC. Thus, for the first time, the ratio k p + /k p  ≅ 1.5 was found directly. The mechanism of the higher efficiency of non-photochemical quenching comparing to photochemical quenching is discussed.

A New Airborne Lidar for Remote Sensing of Canopy Fluorescence and Vertical Profile

NASA Astrophysics Data System (ADS)

Ounis, A.; Bach, J.; Mahjoub, A.; Daumard, F.; Moya, I.; Goulas, Y.

2016-06-01

We report the development of a new lidar system for airborne remote sensing of chlorophyll fluorescence (ChlF) and vertical profile of canopies. By combining laserinduced fluorescence (LIF), sun-induced fluorescence (SIF) and canopy height distribution, the new instrument will low the simultaneous assessment of gross primary production (GPP), photosynthesis efficiency and above ground carbon stocks. Technical issues of the lidar development are discussed and expected performances are presented.

FRET Studies Between CdTe Capped by Small-Molecule Ligands and Fluorescent Protein

NASA Astrophysics Data System (ADS)

Zhang, Yue; Zhou, Dejian; He, Junhui

2014-12-01

Water-soluble luminescent semiconductor nanocrystals also known as quantum dots (QDs) that have prominent photostability, wide absorption cross sections and tunable narrow emission, have been shown as promising probes in immunoassays. QDs are often used as donors in fluorescence resonance energy transfer (FRET) based sensors using organic dyes or fluorescent proteins as acceptors. Here, the FRET between a QD donor and fluorescent protein acceptors has been studied. The fluorescent protein (FP)mCherry appended with a hexa-histidine-tag could effectively self-assemble onto CdTe to produce small donor-acceptor distances and hence highly efficient FRET (efficiency > 80%) at relatively low FP:CdTe copy numbers (ca.1). Using the Förster dipole-dipole interaction formula, the Förster radius (R0) and respective donor-acceptor distances for the CdTe-FP FRET systems have been calculated. The binding constants (Kd) of the QD-FP systems have also been evaluated by the emission spectra.

Materials That Enhance Efficiency and Radiation Resistance of Solar Cells

NASA Technical Reports Server (NTRS)

Sun, Xiadong; Wang, Haorong

2012-01-01

A thin layer (approximately 10 microns) of a novel "transparent" fluorescent material is applied to existing solar cells or modules to effectively block and convert UV light, or other lower solar response waveband of solar radiation, to visible or IR light that can be more efficiently used by solar cells for additional photocurrent. Meanwhile, the layer of fluorescent coating material remains fully "transparent" to the visible and IR waveband of solar radiation, resulting in a net gain of solar cell efficiency. This innovation alters the effective solar spectral power distribution to which an existing cell gets exposed, and matches the maximum photovoltaic (PV) response of existing cells. By shifting a low PV response waveband (e.g., UV) of solar radiation to a high PV response waveband (e.g. Vis-Near IR) with novel fluorescent materials that are transparent to other solar-cell sensitive wavebands, electrical output from solar cells will be enhanced. This approach enhances the efficiency of solar cells by converting UV and high-energy particles in space that would otherwise be wasted to visible/IR light. This innovation is a generic technique that can be readily implemented to significantly increase efficiencies of both space and terrestrial solar cells, without incurring much cost, thus bringing a broad base of economical, social, and environmental benefits. The key to this approach is that the "fluorescent" material must be very efficient, and cannot block or attenuate the "desirable" and unconverted" waveband of solar radiation (e.g. Vis-NIR) from reaching the cells. Some nano-phosphors and novel organometallic complex materials have been identified that enhance the energy efficiency on some state-of-the-art commercial silicon and thin-film-based solar cells by over 6%.

Fluorescent filtered electrophosphorescence

DOEpatents

Forrest, Stephen; Sun, Yiru; Giebink, Noel; Thompson, Mark E.

2010-08-03

The present invention relates to organic light emitting devices (OLEDs), and more specifically to OLEDS that emit light using a combination of fluorescent emitters and phosphorescent emitters for the efficient utilization of all of the electrically generated excitons.

Fluorescent filtered electrophosphorescence

DOEpatents

Forrest, Stephen R [Princeton, NJ; Sun, Yiru [Princeton, NJ; Giebink, Noel [Princeton, NJ; Thompson, Mark E [Anaheim Hills, CA

2009-01-06

The present invention relates to organic light emitting devices (OLEDs), and more specifically to OLEDS that emit light using a combination of fluorescent emitters and phosphorescent emitters for the efficient utilization of all of the electrically generated excitons.

The development of skin immersion clearing method for increasing of laser exposure efficiency on subcutaneous objects

NASA Astrophysics Data System (ADS)

Kozina, Alexandra M.; Genina, Elina A.; Terentyuk, Georgy S.; Terentyuk, Artem G.; Bashkatov, Alexey N.; Tuchin, Valery V.; Khlebtsov, Boris N.

2012-06-01

In this paper we have studied effect of a hyperosmotic optical clearing agent (OCA), such as polyethylene glycol, on the fluorescence intensity from a target located in subcutaneous area in the model experiments. As a fluorescence agent the nanocomposite including gold nanorods with hematophorphyrin was used. The remitted fluorescent signal traveling to the tissue surface was monitored over time as the tissue was treated with the OCA. The detected fluorescent signal increased as the scattering in tissue samples was substantially reduced. The study has shown how OCA can be used to improve the detected signal at localization of subcutaneous target tissue at the photothermal or photodynamic therapy. Immersion clearing of skin can be also useful for improvement of laser exposure efficiency due to the increasing of light penetration depth.

Design of peptide substrates for nanosecond time-resolved fluorescence assays of proteases: 2,3-diazabicyclo[2.2.2]oct-2-ene as a noninvasive fluorophore.

PubMed

Hennig, Andreas; Florea, Mara; Roth, Doris; Enderle, Thilo; Nau, Werner M

2007-01-15

Fluorescence protease assays were investigated with peptide substrates containing a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) as a fluorescent amino acid. The special characteristic of the fluorophore Dbo is its exceedingly long fluorescence lifetime (ca. 300 ns in water under air), which allows the use of nanosecond time-resolved fluorescence (Nano-TRF) detection to efficiently suppress shorter-lived background emission. In addition, the natural amino acids tryptophan and tyrosine can be employed as intramolecular fluorescence quenchers, which facilitates substrate design. Fourteen synthetic peptide substrates (composed of 2-19 amino acids) and five enzymes (trypsin, pepsin, carboxypeptidase A, leucine aminopeptidase, and chymotrypsin) were investigated and, in all 28 examined combinations, enzymatic activity was detected by monitoring the increase in steady state fluorescence with time and determining the reaction rates as kcat/Km values, which ranged from 0.2 to 80x10(6) M-1 min-1. The results suggest an excellent compatibility of the very small and hydrophilic fluorescent probe Dbo with solid-phase peptide synthesis and the investigated proteases. For all 14 peptides the fluorescence lifetimes before and after enzymatic cleavage were measured and Nano-TRF measurements were performed in 384-well microplates. The fluorescence lifetimes of the different peptides provide the basis for the rational design of Dbo-based fluorescent substrates for protease assays. Measurements in Nano-TRF mode revealed, in addition to efficient suppression of background fluorescence, an increased differentiation between cleaved and uncleaved substrate. The Dbo-based assays can be adapted for high-throughput screening.

Designing an anion-functionalized fluorescent ionic liquid as an efficient and reversible turn-off sensor for detecting SO2.

PubMed

Che, Siying; Dao, Rina; Zhang, Weidong; Lv, Xiaoyu; Li, Haoran; Wang, Congmin

2017-03-30

A novel anion-functionalized fluorescent ionic liquid was designed and prepared, which was capable of capturing sulphur dioxide with high capacity and could also be used as a good colorimetric and fluorescent SO 2 sensor. Compared to conventional fluorescent sensors, this fluorescent ionic liquid did not undergo aggregation-caused quenching or aggregation-induced emission, and the fluorescence was quenched when exposed to SO 2 , and the fluorescence would quench when exposed to SO 2 . The experimental absorption, spectroscopic investigation, and quantum chemical calculations indicated that the quenching of the fluorescence originated from SO 2 physical absorption, not chemical absorption. Furthermore, this fluorescent ionic liquid exhibited high selectivity, good quantification, and excellent reversibility for SO 2 detection, and showed potential for an excellent liquid sensor.

Fundamental limits to superresolution fluorescence microscopy

NASA Astrophysics Data System (ADS)

Small, Alex

2013-02-01

Superresolution fluorescence microscopy techniques such as PALM, STORM, STED, and Structured Illumination Microscopy (SIM) enable imaging of live cells at nanometer resolution. The common theme in all of these techniques is that the diffraction limit is circumvented by controlling the states of fluorescent molecules. Although the samples are labeled very densely (i.e. with spacing much smaller than the Airy distance), not all of the molecules are emitting at the same time. Consequently, one does not encounter overlapping blurs. In the deterministic techniques (STED, SIM) the achievable resolution scales as the wavelength of light divided by the square root of the intensity of a beam used to control the fluorescent state. In the stochastic techniques (PALM, STORM), the achievable resolution scales as the wavelength of light divided by the square root of the number of photons collected. Although these limits arise from very different mechanisms (parabolic beam profiles for STED and SIM, statistics for PALM and STORM), in all cases the resolution scales inversely with the square root of a measure of the number of photons used in the experiment. We have developed a proof that this relationship between resolution and photon count is universal to techniques that control the states of fluorophores using classical light. Our proof encompasses linear and nonlinear optics, as well as computational post-processing techniques for extracting information beyond the diffraction limit. If there are techniques that can achieve a more efficient relationship between resolution and photon count, those techniques will require light exhibiting non-classical correlations.

Phosphor Thermometry at ORNL

NASA Astrophysics Data System (ADS)

Allison, S. W.; Gates, M. R.; Beshears, D. L.; Gillies, G. T.

2003-09-01

Phosphor materials are, by design, capable of efficiently converting excitation energy into fluorescence. The temperature-dependent characteristics of this fluorescence provide the basis for noncontact thermometry. In the past decade this approach has been applied to turbine engine diagnostics, liquid temperature measurements for heat pump research, combustion engine intake valve and piston measurements, galvanneal steel processing, transient thermometry of particle beam targets, and microcantilevers used in MEMS devices. The temperatures involved range from ambient to in excess of 1200 °C. Some of these applications have involved fiber optics for light delivery and/or fluorescence signal collection. In addition to fielding these applications, there has been considerable work in the laboratory aimed at exploring further improvements and adding to the database of temperature-characterized phosphors. The activities involve investigation of short-decay time phosphors for use on imaging surfaces moving at high speeds, measuring and modeling pressure as well as temperature dependence, developing phosphor adhesion methods, developing phase-based data acquisition approaches. A significant advance is that light-emitting diodes can now be used to provide adequate stimulation of fluorescence in some applications. Recently nanophosphors have become available. The spectral properties and, by implication, thermal dependence of these properties change with particle size. This has ramifications that need to be explored. The ways in which such materials can be exploited for micro- and nano-technology are just now being addressed. These applications and developments mentioned above will be surveyed and discussed as well as envisioned future improvements and new uses for this thermometry technique.

Resonance Fluorescence of an InGaAs Quantum Dot in a Planar Cavity Using Orthogonal Excitation and Detection.

PubMed

Chen, Disheng; Lander, Gary R; Flagg, Edward B

2017-10-13

The ability to perform simultaneous resonant excitation and fluorescence detection is important for quantum optical measurements of quantum dots (QDs). Resonant excitation without fluorescence detection - for example, a differential transmission measurement - can determine some properties of the emitting system, but does not allow applications or measurements based on the emitted photons. For example, the measurement of photon correlations, observation of the Mollow triplet, and realization of single photon sources all require collection of the fluorescence. Incoherent excitation with fluorescence detection - for example, above band-gap excitation - can be used to create single photon sources, but the disturbance of the environment due to the excitation reduces the indistinguishability of the photons. Single photon sources based on QDs will have to be resonantly excited to have high photon indistinguishability, and simultaneous collection of the photons will be necessary to make use of them. We demonstrate a method to resonantly excite a single QD embedded in a planar cavity by coupling the excitation beam into this cavity from the cleaved face of the sample while collecting the fluorescence along the sample's surface normal direction. By carefully matching the excitation beam to the waveguide mode of the cavity, the excitation light can couple into the cavity and interact with the QD. The scattered photons can couple to the Fabry-Perot mode of the cavity and escape in the surface normal direction. This method allows complete freedom in the detection polarization, but the excitation polarization is restricted by the propagation direction of the excitation beam. The fluorescence from the wetting layer provides a guide to align the collection path with respect to the excitation beam. The orthogonality of the excitation and detection modes enables resonant excitation of a single QD with negligible laser scattering background.

Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances

PubMed Central

Suzuki, Yoshio; Yokoyama, Kenji

2015-01-01

This review is confined to sensors that use fluorescence to transmit biochemical information. Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors. Parameters that define the application of such sensors include intensity, decay time, anisotropy, quenching efficiency, and luminescence energy transfer. To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades. This review describes the immense variety of fluorescent probes that have been designed for the recognitions of ions, small and large molecules, and their biological applications in terms of intracellular fluorescent imaging techniques. PMID:26095660

Chirped circular dielectric gratings for near-unity collection efficiency from quantum emitters in bulk diamond

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zheng, Jiabao; Liapis, Andreas C.; Chen, Edward H.

Effcient collection of fluorescence from nitrogen vacancy (NV) centers in diamond underlies the spin-dependent optical read-out that is necessary for quantum information processing and enhanced sensing applications. The optical collection effciency from NVs within diamond substrates is limited primarily due to the high refractive index of diamond and the non-directional dipole emission. Here we introduce a light collection strategy based on chirped, circular dielectric gratings that can be fabricated on a bulk diamond substrate to redirect an emitter’s far-field radiation pattern. Using a genetic optimization algorithm, these grating designs achieve 98.9% collection effciency for the NV zero-phonon emission line, collectedmore » from the back surface of the diamond with an objective of aperture 0.9. Across the broadband emission spectrum of the NV (600-800 nm), the chirped grating achieves 82.2% collection e ciency into a numerical aperture of 1.42, corresponding to an oil immersion objective again on the back side of the diamond. Our proposed bulk-dielectric grating structures are applicable to other optically active solid state quantum emitters in high index host materials.« less

Chirped circular dielectric gratings for near-unity collection efficiency from quantum emitters in bulk diamond

DOE PAGES

Zheng, Jiabao; Liapis, Andreas C.; Chen, Edward H.; ...

2017-12-13

Effcient collection of fluorescence from nitrogen vacancy (NV) centers in diamond underlies the spin-dependent optical read-out that is necessary for quantum information processing and enhanced sensing applications. The optical collection effciency from NVs within diamond substrates is limited primarily due to the high refractive index of diamond and the non-directional dipole emission. Here we introduce a light collection strategy based on chirped, circular dielectric gratings that can be fabricated on a bulk diamond substrate to redirect an emitter’s far-field radiation pattern. Using a genetic optimization algorithm, these grating designs achieve 98.9% collection effciency for the NV zero-phonon emission line, collectedmore » from the back surface of the diamond with an objective of aperture 0.9. Across the broadband emission spectrum of the NV (600-800 nm), the chirped grating achieves 82.2% collection e ciency into a numerical aperture of 1.42, corresponding to an oil immersion objective again on the back side of the diamond. Our proposed bulk-dielectric grating structures are applicable to other optically active solid state quantum emitters in high index host materials.« less

A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection.

PubMed

Hung, Tran Quang; Chin, Wai Hoe; Sun, Yi; Wolff, Anders; Bang, Dang Duong

2017-04-15

Solid-phase PCR (SP-PCR) has become increasingly popular for molecular diagnosis and there have been a few attempts to incorporate SP-PCR into lab-on-a-chip (LOC) devices. However, their applicability for on-line diagnosis is hindered by the lack of sensitive and portable on-chip optical detection technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Polarized fluorescence for skin cancer diagnostic with a multi-aperture camera

NASA Astrophysics Data System (ADS)

Kandimalla, Haripriya; Ramella-Roman, Jessica C.

2008-02-01

Polarized fluorescence has shown some promising results in assessment of skin cancer margins. Researchers have used tetracycline and cross polarization imaging for nonmelanoma skin cancer demarcation as well as investigating endogenous skin polarized fluorescence. In this paper we present a new instrument for polarized fluorescence imaging, able to calculate the full fluorescence Stokes vector in one snapshot. The core of our system is a multi-aperture camera constructed with a two by two lenslet array. Three of the lenses have polarizing elements in front of them, oriented at 0°, + 45°and 90° with respect to light source polarization. A flash lamp combined with a polarizer parallel to the source-camera-sample plane and a UV filter is used as an excitation source. A blue filter in front of the camera system is used to collect only the fluorescent emission of interest and filter out the incident light. In-vitro tests of endogenous and exogenous polarized fluorescence on collagen rich material like bovine tendon were performed and Stokes vector of polarized fluorescence calculated. The system has the advantage of eliminating moving artifacts with the collection of different polarization states and stoke vector in a single snap shot.

One-step fabrication of PEGylated fluorescent nanodiamonds through the thiol-ene click reaction and their potential for biological imaging

NASA Astrophysics Data System (ADS)

Huang, Hongye; Liu, Meiying; Tuo, Xun; Chen, Junyu; Mao, Liucheng; Wen, Yuanqing; Tian, Jianwen; Zhou, Naigen; Zhang, Xiaoyong; Wei, Yen

2018-05-01

Over the past years, fluorescent carbon nanoparticles have got growing interest for biological imaging. Fluorescent nanodiamonds (FNDs) are novel fluorescent carbon nanoparticles with multitudinous useful properties, including remarkable fluorescence properties, extremely low toxicity and high refractive index. However, facile preparation of FNDs with designable properties and functions from non-fluorescent detonation nanodiamonds (DNDs) has demonstrated to be challengeable. In this work, we reported for the first time that preparation of Polyethylene glycol (PEG) functionalized FNDs through a one-step thiol-ene click reaction using thiol containing PEG (PEG-SH) as the coating agent. Based on the characterization results, we demonstrated that PEG-SH could be efficiently introduced on DNDs to obtain FNDs through the thiol-ene click chemistry. The resultant FND-PEG composites showed high water dispersibility, strong fluorescence and low cytotoxicity. Moreover, FND-PEG composites could be internalized by cells and displayed good cell dyeing performance. All of these features implied that FND-PEG composites are of great potential for biological imaging. Taken together, a facile one-step strategy based on the one-step thiol-ene click reaction has been developed for efficient preparation of FND-PEG composites from non-fluorescent DNDs. The strategy should be also useful for fabrication of many other functional FNDs via using different thiol containing compounds for the universality of thiol-ene click reaction.

A current-assisted CMOS photonic sampler with two taps for fluorescence lifetime sensing

NASA Astrophysics Data System (ADS)

Ingelberts, H.; Kuijk, M.

2016-04-01

Imaging based on fluorescence lifetime is becoming increasingly important in medical and biological applications. State-of- the-art fluorescence lifetime microscopes either use bulky and expensive gated image intensifiers coupled to a CCD or single-photon detectors in a slow scanning setup. Numerous attempts are being made to create compact, cost-effective all- CMOS imagers for fluorescence lifetime sensing. Single-photon avalanche diode (SPAD) imagers can have very good timing resolution and noise characteristics but have low detection efficiency. Another approach is to use CMOS imagers based on demodulation detectors. These imagers can be either very fast or very efficient but it remains a challenge to combine both characteristics. Recently we developed the current-assisted photonic sampler (CAPS) to tackle these problems and in this work, we present a new CAPS with two detection taps that can sample a fluorescence decay in two time windows. In the case of mono-exponential decays, two windows provide enough information to resolve the lifetime. We built an electro-optical setup to characterize the detector and use it for fluorescence lifetime measurements. It consists of a supercontinuum pulsed laser source, an optical system to focus light into the detector and picosecond timing electronics. We describe the structure and operation of the two-tap CAPS and provide basic characterization of the speed performance at multiple wavelengths in the visible and near-infrared spectrum. We also record fluorescence decays of different visible and NIR fluorescent dyes and provide different methods to resolve the fluorescence lifetime.

Combining CPP-ACP with fluoride: a synergistic remineralization potential of artificially demineralized enamel or not?

NASA Astrophysics Data System (ADS)

El-Sayad, I. I.; Sakr, A. K.; Badr, Y. A.

2008-08-01

Background and objective: Minimal intervention dentistry (MID) calls for early detection and remineralization of initial demineralization. Laser fluorescence is efficient in detecting changes in mineral tooth content. Recaldent is a product of casein phosphopeptide-amorphous calcium phosphate (CPP- ACP) which delivers calcium and phosphate ions to enamel. A new product which also contains fluoride is launched in United States. The remineralizing potential of CPP- ACP per se, or when combined with 0.22% Fl supplied in an oral care gel on artificially demineralised enamel using laser fluorescence was investigated. Methods: Fifteen sound human molars were selected. Mesial surfaces were tested using He-Cd laser beam at 441.5nm with 18mW power as excitation source on a suitable set-up based on Spex 750 M monochromator provided with PMT for detection of collected auto-fluorescence from sound enamel. Mesial surfaces were subjected to demineralization for ten days. The spectra from demineralized enamel were measured. Teeth were then divided according to the remineralizing regimen into three groups: group I recaldent per se, group II recaldent combined with fluoride gel and group III artificial saliva as a positive control. After following these protocols for three weeks, the spectra from remineralized enamel from the three groups were measured. The spectra of enamel auto-fluorescence were recorded and normalized to peak intensity at about 540 nm to compare between spectra from sound, demineralized and remineralized enamel surfaces. Results: A slight red shift was noticed in spectra from demineralized enamel, while a blue shift may occur in remineralized enamel. Group II showed the highest remineralizing potential. Conclusions: Combining fluoride with CPP-ACP had a synergistic effect on enamel remineralization. In addition, laser auto-fluorescence is an accurate technique for assessment of changes in tooth enamel minerals.

Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

NASA Astrophysics Data System (ADS)

Xu, Dongli; Peng, Leilei

2016-03-01

Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

Efficient globally optimal segmentation of cells in fluorescence microscopy images using level sets and convex energy functionals.

PubMed

Bergeest, Jan-Philip; Rohr, Karl

2012-10-01

In high-throughput applications, accurate and efficient segmentation of cells in fluorescence microscopy images is of central importance for the quantification of protein expression and the understanding of cell function. We propose an approach for segmenting cell nuclei which is based on active contours using level sets and convex energy functionals. Compared to previous work, our approach determines the global solution. Thus, the approach does not suffer from local minima and the segmentation result does not depend on the initialization. We consider three different well-known energy functionals for active contour-based segmentation and introduce convex formulations of these functionals. We also suggest a numeric approach for efficiently computing the solution. The performance of our approach has been evaluated using fluorescence microscopy images from different experiments comprising different cell types. We have also performed a quantitative comparison with previous segmentation approaches. Copyright © 2012 Elsevier B.V. All rights reserved.

Two-photon excited fluorescence from a pseudoisocyanine-attached gold-coated tip via a thin tapered fiber under a weak continuous wave excitation.

PubMed

Ren, Fang; Takashima, Hideaki; Tanaka, Yoshito; Fujiwara, Hideki; Sasaki, Keiji

2013-11-18

A simple tapered fiber based photonic-plasmonic hybrid nanostructure composed of a thin tapered fiber and a pseudoisocyanine (PIC)-attached Au-coated tip was demonstrated. Using this simple hybrid nanostructure, we succeeded in observing two-photon excited fluorescence from the PIC dye molecules under a weak continuous wave excitation condition. From the results of the tip-fiber distance dependence and excitation polarization dependence, we found that using a thin tapered fiber and an Au-coated tip realized efficient coupling of the incident light (~95%) and LSP excitation at the Au-coated tip, suggesting the possibility of efficiently inducing two-photon excited fluorescence from the PIC dye molecules attached on the Au-coated tip. This simple photonic-plasmonic hybrid system is one of the promising tools for single photon sources, highly efficient plasmonic sensors, and integrated nonlinear plasmonic devices.

Achieving 100% Efficient Postcolumn Hydride Generation for As Speciation Analysis by Atomic Fluorescence Spectrometry.

PubMed

Marschner, Karel; Musil, Stanislav; Dědina, Jiří

2016-04-05

An experimental setup consisting of a flow injection hydride generator coupled to an atomic fluorescence spectrometer was optimized in order to generate arsanes from tri- and pentavalent inorganic arsenic species (iAs(III), iAs(V)), monomethylarsonic acid (MAs(V)), and dimethylarsinic acid (DMAs(V)) with 100% efficiency with the use of only HCl and NaBH4 as the reagents. The optimal concentration of HCl was 2 mol L(-1); the optimal concentration of NaBH4 was 2.5% (m/v), and the volume of the reaction coil was 8.9 mL. To prevent excessive signal noise due to fluctuations of hydride supply to an atomizer, a new design of a gas-liquid separator was implemented. The optimized experimental setup was subsequently interfaced to HPLC and employed for speciation analysis of arsenic. Two chromatography columns were tested: (i) ion-pair chromatography and (ii) ion exchange chromatography. The latter offered much better results for human urine samples without a need for sample dilution. Due to the equal hydride generation efficiency (and thus the sensitivities) of all As species, a single species standardization by DMAs(V) standard was feasible. The limits of detection for iAs(III), iAs(V), MAs(V), and DMAs(V) were 40, 97, 57, and 55 pg mL(-1), respectively. Accuracy of the method was tested by the analysis of the standard reference material (human urine NIST 2669), and the method was also verified by the comparative analyses of human urine samples collected from five individuals with an independent reference method.

Highly efficient deep-blue organic light emitting diode with a carbazole based fluorescent emitter

NASA Astrophysics Data System (ADS)

Sahoo, Snehasis; Dubey, Deepak Kumar; Singh, Meenu; Joseph, Vellaichamy; Thomas, K. R. Justin; Jou, Jwo-Huei

2018-04-01

High efficiency deep-blue emission is essential to realize energy-saving, high-quality display and lighting applications. We demonstrate here a deep-blue organic light emitting diode using a novel carbazole based fluorescent emitter 7-[4-(diphenylamino)phenyl]-9-(2-ethylhexyl)-9H-carbazole-2-carbonitrile (JV234). The solution processed resultant device shows a maximum luminance above 1,750 cd m-2 and CIE coordinates (0.15,0.06) with a 1.3 lm W-1 power efficiency, 2.0 cd A-1 current efficiency, and 4.1% external quantum efficiency at 100 cd m-2. The resulting deep-blue emission enables a greater than 100% color saturation. The high efficiency may be attributed to the effective host-to-guest energy transfer, suitable device architecture facilitating balanced carrier injection and low doping concentration preventing efficiency roll-off caused by concentration quenching.

Efficient expression of green fluorescent protein (GFP) mediated by a chimeric promoter in Chlamydomonas reinhardtii

NASA Astrophysics Data System (ADS)

Wu, Jinxia; Hu, Zhangli; Wang, Chaogang; Li, Shuangfei; Lei, Anping

2008-08-01

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein (GFP) was expressed in C. reinhardtii under the control of promoters: RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae: RBCS2 mediated strain Tran-I and HSP70A-RBCS2 mediated strain Tran-II. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-II was at least double of that in Tran-I. In addition, a threefold increase of GFP in Tran-II was induced by heat shock at 40°C. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.

A pH sensing system using fluorescence-based fibre optical sensor capable of small volume sample measurement

NASA Astrophysics Data System (ADS)

Deng, Shijie; McAuliffe, Michael A. P.; Salaj-Kosla, Urszula; Wolfe, Raymond; Lewis, Liam; Huyet, Guillaume

2017-02-01

In this work, a low cost optical pH sensing system that allows for small volume sample measurements was developed. The system operates without the requirement of laboratory instruments (e.g. laser source, spectrometer and CCD camera), this lowers the cost and enhances the portability. In the system, an optical arrangement employing a dichroic filter was used which allows the excitation and emission light to be transmitted using a single fibre thus improving the collection efficiency of the fluorescence signal and also the ability of inserting measurement. The pH sensor in the system uses bromocresol purple as the indicator which is immobilised by sol-gel technology through a dip-coating process. The sensor material was coated on the tip of a 1 mm diameter optical fibre which makes it possible for inserting into very small volume samples to measure the pH. In the system, a LED with a peak emission wavelength of 465 nm is used as the light source and a silicon photo-detector is used to detect the uorescence signal. Optical filters are applied after the LED and in front of the photo-detector to separate the excitation and emission light. The fluorescence signal collected is transferred to a PC through a DAQ and processed by a Labview-based graphic-user-interface (GUI). Experimental results show that the system is capable of sensing pH values from 5.3 to 8.7 with a linear response of R2=0.969. Results also show that the response times for a pH changes from 5.3 to 8.7 is approximately 150 s and for a 0.5 pH changes is approximately 50 s.

Efficient L1 regularization-based reconstruction for fluorescent molecular tomography using restarted nonlinear conjugate gradient.

PubMed

Shi, Junwei; Zhang, Bin; Liu, Fei; Luo, Jianwen; Bai, Jing

2013-09-15

For the ill-posed fluorescent molecular tomography (FMT) inverse problem, the L1 regularization can protect the high-frequency information like edges while effectively reduce the image noise. However, the state-of-the-art L1 regularization-based algorithms for FMT reconstruction are expensive in memory, especially for large-scale problems. An efficient L1 regularization-based reconstruction algorithm based on nonlinear conjugate gradient with restarted strategy is proposed to increase the computational speed with low memory consumption. The reconstruction results from phantom experiments demonstrate that the proposed algorithm can obtain high spatial resolution and high signal-to-noise ratio, as well as high localization accuracy for fluorescence targets.

Spectroscopic probing of ortho-nitrophenol localization in phospholipid bilayers.

PubMed

Sánchez, Julieta M; Turina, Anahí del V; Perillo, María A

2007-11-12

In the present study we estimated the localization of ortho-nitrophenol (ONP) within model membranes through its efficiency to quench and to modify the anisotropy of DPH and TMA-DPH fluorescence. These fluorescent probes are known to sense the hydrocarbon core and the polar head group region of membranes, respectively. TMA-DPH fluorescence in MLVs was more efficiently quenched than DPH (K(q,TMA-DPH)=2.36 and K(q,DPH)=1.07 mM ns(-1)). Moreover, these results demonstrated the interfacial localization of ONP and may contribute to understand membrane-mediated mechanisms of ONP-induced toxicity and the behavior of ONP as a product of several enzymatic reactions occurring in the presence of lipid-water interfaces.

Fast globally optimal segmentation of cells in fluorescence microscopy images.

PubMed

Bergeest, Jan-Philip; Rohr, Karl

2011-01-01

Accurate and efficient segmentation of cells in fluorescence microscopy images is of central importance for the quantification of protein expression in high-throughput screening applications. We propose a new approach for segmenting cell nuclei which is based on active contours and convex energy functionals. Compared to previous work, our approach determines the global solution. Thus, the approach does not suffer from local minima and the segmentation result does not depend on the initialization. We also suggest a numeric approach for efficiently computing the solution. The performance of our approach has been evaluated using fluorescence microscopy images of different cell types. We have also performed a quantitative comparison with previous segmentation approaches.

Design of ortho-Substituted Donor-Acceptor Molecules as Highly Efficient Green Thermally Activated Delayed Fluorescent Emitters

NASA Astrophysics Data System (ADS)

Cha, Jae-Ryung; Gong, Myoung-Seon; Lee, Tak Jae; Ha, Tae Hoon; Lee, Chil Won

2018-04-01

The ortho-substituted donor-acceptor molecules 2-(4,6-diphenyl-1, 3, 5-triazin-2-yl)- N,Ndiphenylaniline (DPA- o-Trz) and 2-(4,6-diphenyl-1, 3, 5-triazine-2-yl)- N,N-di- p-tolylaniline (MPA- o-Trz) were designed, synthesized, and found to exhibit green fluorescence characteristics. Notably, the singlet-triplet energy gap was less than 0.1 eV, indicating that reverse intersystem crossing gave rise to thermally activated delayed fluorescence (TADF). The organic light-emitting device performance of MPA- o-Trz showed a high external quantum efficiency of 16.3% and good color stability from 0.1 cd/m2 to 5000 cd/m2.

Fluorescence lifetime imaging of skin cancer

NASA Astrophysics Data System (ADS)

Patalay, Rakesh; Talbot, Clifford; Munro, Ian; Breunig, Hans Georg; König, Karsten; Alexandrov, Yuri; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Chris

2011-03-01

Fluorescence intensity imaging and fluorescence lifetime imaging microscopy (FLIM) using two photon microscopy (TPM) have been used to study tissue autofluorescence in ex vivo skin cancer samples. A commercially available system (DermaInspect®) was modified to collect fluorescence intensity and lifetimes in two spectral channels using time correlated single photon counting and depth-resolved steady state measurements of the fluorescence emission spectrum. Uniquely, image segmentation has been used to allow fluorescence lifetimes to be calculated for each cell. An analysis of lifetime values obtained from a range of pigmented and non-pigmented lesions will be presented.

Photochemical synthesis and photophysical properties of coumarins bearing extended polyaromatic rings studied by emission and transient absorption measurements.

PubMed

Yamaji, Minoru; Hakoda, Yuma; Okamoto, Hideki; Tani, Fumito

2017-04-12

We prepared a variety of coumarin derivatives having expanded π-electron systems along the direction crossing the C 3 -C 4 bond of the coumarin skeleton via a photochemical cyclization process and investigated their photophysical features as a function of the number (n) of the added benzene rings based on emission and transient absorption measurements. Upon increasing n, the fluorescence quantum yields of the π-extended coumarins increased. Expanding the π-electron system on the C 3 -C 4 bond of the coumarin skeleton was found to be efficient for increasing the fluorescence ability more than that on the C 7 -C 8 bond. Introducing the methoxy group at the 7-position was also efficient for enhancing the fluorescence quantum yield and rate of the expanded coumarins. The non-radiative process from the fluorescence state was not substantially influenced by the expanded π-electron system. The competitive process with the fluorescence was found to be intersystem crossing to the triplet state based on the observations of the triplet-triplet absorption. The effects of the expanded π-electron systems on the fluorescence ability were investigated with the aid of TD-DFT calculations.

A coumarin-based "turn-on" fluorescent sensor for the determination of Al3+: single crystal X-ray structure and cell staining properties.

PubMed

Guha, Subarna; Lohar, Sisir; Sahana, Animesh; Banerjee, Arnab; Safin, Damir A; Babashkina, Maria G; Mitoraj, Mariusz P; Bolte, Michael; Garcia, Yann; Mukhopadhyay, Subhra Kanti; Das, Debasis

2013-07-28

An efficient Al(3+) receptor, 6-(2-hydroxybenzylideneamino)-2H-chromen-2-one (HBC), has been synthesized by condensing salicylaldehyde with 6-aminocoumarin. The molecular structure of HBC has been determined by a single crystal X-ray analysis. It was established that in the presence of Al(3+), HBC shows 25 fold enhancement of fluorescence intensity which might be attributed to the chelation-enhanced fluorescence (CHEF) process. HBC binds Al(NO3)3 in a 1 : 1 stoichiometry with a binding constant (K) of 7.9 × 10(4) M(-1). Fe(3+) and Mn(2+) quench the emission intensity of the [HBC + Al(3+)] system to an insignificant extent at a concentration 10 times higher compared to that of Al(3+). HBC is highly efficient in the detection of intracellular Al(3+) under a fluorescence microscope.

Imaging workflow and calibration for CT-guided time-domain fluorescence tomography

PubMed Central

Tichauer, Kenneth M.; Holt, Robert W.; El-Ghussein, Fadi; Zhu, Qun; Dehghani, Hamid; Leblond, Frederic; Pogue, Brian W.

2011-01-01

In this study, several key optimization steps are outlined for a non-contact, time-correlated single photon counting small animal optical tomography system, using simultaneous collection of both fluorescence and transmittance data. The system is presented for time-domain image reconstruction in vivo, illustrating the sensitivity from single photon counting and the calibration steps needed to accurately process the data. In particular, laser time- and amplitude-referencing, detector and filter calibrations, and collection of a suitable instrument response function are all presented in the context of time-domain fluorescence tomography and a fully automated workflow is described. Preliminary phantom time-domain reconstructed images demonstrate the fidelity of the workflow for fluorescence tomography based on signal from multiple time gates. PMID:22076264

Application of air ions for bacterial de-colonization in air filters contaminated by aerosolized bacteria.

PubMed

Kim, Yang Seon; Yoon, Ki Young; Park, Jae Hong; Hwang, Jungho

2011-01-15

We aerosolized the Escherichia coli (E. coli) and Staphylococcus epidermidis (S. epidermidis) bacteria and collected them on membrane filters. Then we generated air ions by applying a high voltage to a carbon fiber tip and applied them to the contaminated filters. The antibacterial efficiency was not significantly affected by the bacteria being Gram-positive or Gram-negative, however, negative ions showed a lower antibacterial efficiency than positive ions to both E. coli and S. epidermidis, even though the concentration of negative air ions was much higher than that of positive air ions. With a field emission scanning electron microscope (FE-SEM) images and fluorescence microscopy images using a LIVE/DEAD BacLight Bacterial Viability Kit, electrostatic disruption of the bacteria was found to be the dominant antibacterial effect. Copyright © 2010 Elsevier B.V. All rights reserved.

Baculovirus-mediated gene transfer in butterfly wings in vivo: an efficient expression system with an anti-gp64 antibody.

PubMed

Dhungel, Bidur; Ohno, Yoshikazu; Matayoshi, Rie; Otaki, Joji M

2013-03-25

Candidate genes for color pattern formation in butterfly wings have been known based on gene expression patterns since the 1990s, but their functions remain elusive due to a lack of a functional assay. Several methods of transferring and expressing a foreign gene in butterfly wings have been reported, but they have suffered from low success rates or low expression levels. Here, we developed a simple, practical method to efficiently deliver and express a foreign gene using baculovirus-mediated gene transfer in butterfly wings in vivo. A recombinant baculovirus containing a gene for green fluorescent protein (GFP) was injected into pupae of the blue pansy butterfly Junonia orithya (Nymphalidae). GFP fluorescence was detected in the pupal wings and other body parts of the injected individuals three to five days post-injection at various degrees of fluorescence. We obtained a high GFP expression rate at relatively high virus titers, but it was associated with pupal death before color pattern formation in wings. To reduce the high mortality rate caused by the baculovirus treatment, we administered an anti-gp64 antibody, which was raised against baculovirus coat protein gp64, to infected pupae after the baculovirus injection. This treatment greatly reduced the mortality rate of the infected pupae. GFP fluorescence was observed in pupal and adult wings and other body parts of the antibody-treated individuals at various degrees of fluorescence. Importantly, we obtained completely developed wings with a normal color pattern, in which fluorescent signals originated directly from scales or the basal membrane after the removal of scales. GFP fluorescence in wing tissues spatially coincided with anti-GFP antibody staining, confirming that the fluorescent signals originated from the expressed GFP molecules. Our baculovirus-mediated gene transfer system with an anti-gp64 antibody is reasonably efficient, and it can be an invaluable tool to transfer, express, and functionally examine foreign genes in butterfly wings and also in other non-model insect systems.

Fluorescent minerals, a review

USGS Publications Warehouse

Modreski, P.J.; Aumente-Modreski, R.

1996-01-01

Fluorescent minerals are more than just an attractive novelty, and collecting them is a speciality for thousands of individuals who appreciate their beauty, rarity, and scientific value. Fluorescent properties can be used as an aid to mineral identification, locality determination, and distinction between natural and synthetic gemstones. This article gives an overview of those aspects of fluorescence that are of most interest to collectors, hobbyists, and mineralogists. -from Authors

Solid state laser media driven by remote nuclear powered fluorescence

DOEpatents

Prelas, Mark A.

1992-01-01

An apparatus is provided for driving a solid state laser by a nuclear powered fluorescence source which is located remote from the fluorescence source. A nuclear reaction produced in a reaction chamber generates fluorescence or photons. The photons are collected from the chamber into a waveguide, such as a fiber optic waveguide. The waveguide transports the photons to the remote laser for exciting the laser.

Highly efficient blue and warm white organic light-emitting diodes with a simplified structure

NASA Astrophysics Data System (ADS)

Li, Xiang-Long; Ouyang, Xinhua; Chen, Dongcheng; Cai, Xinyi; Liu, Ming; Ge, Ziyi; Cao, Yong; Su, Shi-Jian

2016-03-01

Two blue fluorescent emitters were utilized to construct simplified organic light-emitting diodes (OLEDs) and the remarkable difference in device performance was carefully illustrated. A maximum current efficiency of 4.84 cd A-1 (corresponding to a quantum efficiency of 4.29%) with a Commission Internationale de l’Eclairage (CIE) coordinate of (0.144, 0.127) was achieved by using N,N-diphenyl-4″-(1-phenyl-1H-benzo[d]imidazol-2-yl)-[1, 1‧:4‧, 1″-terphenyl]-4-amine (BBPI) as a non-doped emission layer of the simplified blue OLEDs without carrier-transport layers. In addition, simplified fluorescent/phosphorescent (F/P) hybrid warm white OLEDs without carrier-transport layers were fabricated by utilizing BBPI as (1) the blue emitter and (2) the host of a complementary yellow phosphorescent emitter (PO-01). A maximum current efficiency of 36.8 cd A-1 and a maximum power efficiency of 38.6 lm W-1 were achieved as a result of efficient energy transfer from the host to the guest and good triplet exciton confinement on the phosphorescent molecules. The blue and white OLEDs are among the most efficient simplified fluorescent blue and F/P hybrid white devices, and their performance is even comparable to that of most previously reported complicated multi-layer devices with carrier-transport layers.

Highly Efficient Multiple-Anchored Fluorescent Probe for the Detection of Aniline Vapor Based on Synergistic Effect: Chemical Reaction and PET.

PubMed

Jiao, Zinuo; Zhang, Yu; Xu, Wei; Zhang, Xiangtao; Jiang, Haibo; Wu, Pengcheng; Fu, Yanyan; He, Qingguo; Cao, Huimin; Cheng, Jiangong

2017-05-26

A multiple-anchored fluorescent probe ((((hexane-1,6-diylbis(2,7-bis(4-formyl)-phenyl)-9H-fluorine-9,9-diyl))-bis(hexane-6,1-diyl))-bis(9H-carbazole-9,3,6-triyl))-tetrakis(benzene-4,1-diyl))-tetraformyl-(8FP-2F) with eight aldehyde groups was designed and synthesized. The molecule has four branches and highly twisted structure. Furthermore, it tends to self-assemble into nanospheres, which is beneficial for gaseous analyte penetration and high fluorescence quantum efficiency. Among gaseous analytes, detection of aniline vapor is extraordinarily important in the control of environmental issues and human diseases. Herein, 8FP-2F was introduced to detect aniline vapor with distinguished sensitivity and selectivity via simple Schiff base reaction at room temperature. After exposure to saturate aniline vapor, the 89% fluorescence of 8FP-2F was quenched in 50 s and the detection limit was as low as 3 ppb. Further study showed the suitable HOMO/LUMO energy levels and matched orbital symmetry between probe and aniline molecules ensured chemical reaction and PET process work together. The synergistic effect resulted in a significant sensing performance and fluorescence quenching toward aniline vapor. Moreover, the multiple active sites structure of 8FP-2F means it could be applied for constructing many interesting structures and highly efficient organic optoelectronic functional materials.

Comparison of flow cytometry, fluorescence microscopy and spectrofluorometry for analysis of gene electrotransfer efficiency.

PubMed

Marjanovič, Igor; Kandušer, Maša; Miklavčič, Damijan; Keber, Mateja Manček; Pavlin, Mojca

2014-12-01

In this study, we compared three different methods used for quantification of gene electrotransfer efficiency: fluorescence microscopy, flow cytometry and spectrofluorometry. We used CHO and B16 cells in a suspension and plasmid coding for GFP. The aim of this study was to compare and analyse the results obtained by fluorescence microscopy, flow cytometry and spectrofluorometry and in addition to analyse the applicability of spectrofluorometry for quantifying gene electrotransfer on cells in a suspension. Our results show that all the three methods detected similar critical electric field strength, around 0.55 kV/cm for both cell lines. Moreover, results obtained on CHO cells showed that the total fluorescence intensity and percentage of transfection exhibit similar increase in response to increase electric field strength for all the three methods. For B16 cells, there was a good correlation at low electric field strengths, but at high field strengths, flow cytometer results deviated from results obtained by fluorescence microscope and spectrofluorometer. Our study showed that all the three methods detected similar critical electric field strengths and high correlations of results were obtained except for B16 cells at high electric field strengths. The results also demonstrated that flow cytometry measures higher values of percentage transfection compared to microscopy. Furthermore, we have demonstrated that spectrofluorometry can be used as a simple and consistent method to determine gene electrotransfer efficiency on cells in a suspension.

What limits photosynthetic energy conversion efficiency in nature? Lessons from the oceans.

PubMed

Falkowski, Paul G; Lin, Hanzhi; Gorbunov, Maxim Y

2017-09-26

Constraining photosynthetic energy conversion efficiency in nature is challenging. In principle, two yield measurements must be made simultaneously: photochemistry, fluorescence and/or thermal dissipation. We constructed two different, extremely sensitive and precise active fluorometers: one measures the quantum yield of photochemistry from changes in variable fluorescence, the other measures fluorescence lifetimes in the picosecond time domain. By deploying the pair of instruments on eight transoceanic cruises over six years, we obtained over 200 000 measurements of fluorescence yields and lifetimes from surface waters in five ocean basins. Our results revealed that the average quantum yield of photochemistry was approximately 0.35 while the average quantum yield of fluorescence was approximately 0.07. Thus, closure on the energy budget suggests that, on average, approximately 58% of the photons absorbed by phytoplankton in the world oceans are dissipated as heat. This extraordinary inefficiency is associated with the paucity of nutrients in the upper ocean, especially dissolved inorganic nitrogen and iron. Our results strongly suggest that, in nature, most of the time, most of the phytoplankton community operates at approximately half of its maximal photosynthetic energy conversion efficiency because nutrients limit the synthesis or function of essential components in the photosynthetic apparatus.This article is part of the themed issue 'Enhancing photosynthesis in crop plants: targets for improvement'. © 2017 The Author(s).

Controlling Synergistic Oxidation Processes for Efficient and Stable Blue Thermally Activated Delayed Fluorescence Devices.

PubMed

Cui, Lin-Song; Deng, Ya-Li; Tsang, Daniel Ping-Kuen; Jiang, Zuo-Quan; Zhang, Qisheng; Liao, Liang-Sheng; Adachi, Chihaya

2016-09-01

Efficient sky-blue organic light-emitting diodes (OLEDs) employing thermally activated delayed fluorescence (TADF) display a three orders of magnitude increase in lifetime, which is superior to those of controlled phosphorescent OLEDs used in this study. The combination of electro-oxidation and photo-oxidation of the TADF emitters in their triplet excited-states is suppressed through molecule design and device engineering. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A novel near-infrared nanomaterial with high quantum efficiency and its applications in real time in vivo imaging

NASA Astrophysics Data System (ADS)

Cui, X. X.; Fan, Q.; Shi, S. J.; Wen, W. H.; Chen, D. F.; Guo, H. T.; Xu, Y. T.; Gao, F.; Nie, R. Z.; Ford, Harold D.; Tang, Gordon H.; Hou, C. Q.; Peng, B.

2018-05-01

Fluorescence imaging signal is severely limited by the quantum efficiency and emission wavelength. To overcome these challenges, novel NIR-emitting K5NdLi2F10 nanoparticles under NIR excitation was introduced as fluorescence imaging probe for the first time. The photostability of K5NdLi2F10 nanoparticles in the water, phosphate buffer saline, fetal bovine serum and living mice was investigated. The fluorescence signal was detected with depths of 3.5 and 2.0 cm in phantom and pork tissue, respectively. Fluorescence spectrum with a significant signal-to-background ratio of 10:1 was captured in living mice. Moreover, clear NIR images were virtualized for the living mice after intravenous injection. The imaging ability of nanoparticles in tumor-beard mice were evaluated, the enrichment of K5NdLi2F10 nanoparticles in tumor site due to the enhanced permeability and retention effect was confirmed. The systematic studies of toxicity, bio-distribution and in-vivo dynamic imaging suggest that these materials give high biocompatibility and low toxicity. These NIR-emitting nanoparticles with high quantum efficiency, high penetration and low toxicity might facilitate tumor identification in deep tissues more sensitively.

A novel near-infrared nanomaterial with high quantum efficiency and its applications in real time in vivo imaging.

PubMed

Cui, X X; Fan, Q; Shi, S J; Wen, W H; Chen, D F; Guo, H T; Xu, Y T; Gao, F; Nie, R Z; Ford, Harold D; Tang, Gordon H; Hou, C Q; Peng, B

2018-05-18

Fluorescence imaging signal is severely limited by the quantum efficiency and emission wavelength. To overcome these challenges, novel NIR-emitting K 5 NdLi 2 F 10 nanoparticles under NIR excitation was introduced as fluorescence imaging probe for the first time. The photostability of K 5 NdLi 2 F 10 nanoparticles in the water, phosphate buffer saline, fetal bovine serum and living mice was investigated. The fluorescence signal was detected with depths of 3.5 and 2.0 cm in phantom and pork tissue, respectively. Fluorescence spectrum with a significant signal-to-background ratio of 10:1 was captured in living mice. Moreover, clear NIR images were virtualized for the living mice after intravenous injection. The imaging ability of nanoparticles in tumor-beard mice were evaluated, the enrichment of K 5 NdLi 2 F 10 nanoparticles in tumor site due to the enhanced permeability and retention effect was confirmed. The systematic studies of toxicity, bio-distribution and in-vivo dynamic imaging suggest that these materials give high biocompatibility and low toxicity. These NIR-emitting nanoparticles with high quantum efficiency, high penetration and low toxicity might facilitate tumor identification in deep tissues more sensitively.

Development of the Concept of Recycling of Light

NASA Astrophysics Data System (ADS)

Harmer, Brian

Environmental and economic issues are the highlights of any new product or system created today. The efficient use of energy helps satisfy both of these concerns as a reduction in energy consumption contributes to a reduction both in fuel consumption and carbon emissions. Illumination efficiency has been one of the main areas of research as luminaires are one of the largest consumers of electricity in the world. The incandescent bulb is one of the oldest pieces of technology still used today, but is being phased out as compact fluorescent lamps and LED light sources have a much lower power consumption for the same amount of light emission. However, the light source design, while very important, is not the only way to improve the efficiency of an illumination system. This thesis proposes a new concept, the recycling of light (ROL). The ROL system collects, transports, and emits unused light from one area to another through the use of optical fibers. To find an optimal ROL system, many variables need to be accounted for. This thesis covers the effect of different luminaires on light collection areas. The collection area for the ROL system needs to be placed in the areas of a room that are of little or no importance, but still receive light, such as the ceiling or the upper section of the walls. The fiber-to-source distance and offset effects on fiber emission are investigated, as well as the length and type of the optical fibers. Additionally, this thesis looks at the possibility of beveling optical fiber ends to be used as a focusing mechanism for the ROL system.

Collective fluorescence and decoherence of a few nearly identical quantum dots

NASA Astrophysics Data System (ADS)

Sitek, Anna; Machnikowski, Paweł

2007-01-01

We study the collective interaction of excitons in closely spaced artificial molecules and arrays of nearly identical quantum dots with the electromagnetic modes. We discuss how collective fluorescence builds up in the presence of a small mismatch of the transition energy. We show that a superradiant state of a single exciton in a molecule of two dots with realistic energy mismatch undergoes a two-rate decay. We also analyze the stability of subdecoherent states for nonidentical systems.

[MODIS Investigation

NASA Technical Reports Server (NTRS)

Abbott, Mark R.

1998-01-01

The objectives of the last six months were: (1) Revise the algorithms for the Fluorescence Line Height (FLH) and Chlorophyll Fluorescence Efficiency (CFE) products, especially the data quality flags; (2) Revise the MOCEAN validation plan; (3) Deploy and recover bio-optical instrumentation at the Hawaii Ocean Time-series (HOT) site as part of the Joint Global Ocean Flux Study (JGOFS); (4) Prepare for field work in the Antarctic Polar Frontal Zone as part of JGOFS; (5) Submit manuscript on bio-optical time scales as estimated from Lagrangian drifters; (6) Conduct chemostat experiments on fluorescence; (7) Interface with the Global Imager (GLI) science team; and (8) Continue development of advanced data system browser. We are responsible for the delivery of two at-launch products for AM-1: Fluorescence line height (FLH) and chlorophyll fluorescence efficiency (CFE). We also considered revising the input chlorophyll, which is used to determine the degree of binning. We have refined the quality flags for the Version 2 algorithms. We have acquired and installed a Silicon Graphics Origin 200. We are working with the University of Miami team to develop documentation that will describe how the MODIS ocean components are linked together.

Systemic Propagation of a Fluorescent Infectious Clone of a Polerovirus Following Inoculation by Agrobacteria and Aphids

PubMed Central

Boissinot, Sylvaine; Pichon, Elodie; Sorin, Céline; Piccini, Céline; Scheidecker, Danièle; Ziegler-Graff, Véronique; Brault, Véronique

2017-01-01

A fluorescent viral clone of the polerovirus Turnip yellows virus (TuYV) was engineered by introducing the Enhanced Green Fluorescent Protein (EGFP) sequence into the non-structural domain sequence of the readthrough protein, a minor capsid protein. The resulting recombinant virus, referred to as TuYV-RTGFP, was infectious in several plant species when delivered by agroinoculation and invaded efficiently non-inoculated leaves. As expected for poleroviruses, which infect only phloem cells, the fluorescence emitted by TuYV-RTGFP was restricted to the vasculature of infected plants. In addition, TuYV-RTGFP was aphid transmissible and enabled the observation of the initial sites of infection in the phloem after aphid probing in epidermal cells. The aphid-transmitted virus moved efficiently to leaves distant from the inoculation sites and importantly retained the EGFP sequence in the viral genome. This work reports on the first engineered member in the Luteoviridae family that can be visualized by fluorescence emission in systemic leaves of different plant species after agroinoculation or aphid transmission. PMID:28661469

Systemic Propagation of a Fluorescent Infectious Clone of a Polerovirus Following Inoculation by Agrobacteria and Aphids.

PubMed

Boissinot, Sylvaine; Pichon, Elodie; Sorin, Céline; Piccini, Céline; Scheidecker, Danièle; Ziegler-Graff, Véronique; Brault, Véronique

2017-06-29

A fluorescent viral clone of the polerovirus Turnip yellows virus (TuYV) was engineered by introducing the Enhanced Green Fluorescent Protein (EGFP) sequence into the non-structural domain sequence of the readthrough protein, a minor capsid protein. The resulting recombinant virus, referred to as TuYV-RT GFP , was infectious in several plant species when delivered by agroinoculation and invaded efficiently non-inoculated leaves. As expected for poleroviruses, which infect only phloem cells, the fluorescence emitted by TuYV-RT GFP was restricted to the vasculature of infected plants. In addition, TuYV-RT GFP was aphid transmissible and enabled the observation of the initial sites of infection in the phloem after aphid probing in epidermal cells. The aphid-transmitted virus moved efficiently to leaves distant from the inoculation sites and importantly retained the EGFP sequence in the viral genome. This work reports on the first engineered member in the Luteoviridae family that can be visualized by fluorescence emission in systemic leaves of different plant species after agroinoculation or aphid transmission.

CRISPR/Cas9-Mediated Fluorescent Tagging of Endogenous Proteins in Human Pluripotent Stem Cells.

PubMed

Sharma, Arun; Toepfer, Christopher N; Ward, Tarsha; Wasson, Lauren; Agarwal, Radhika; Conner, David A; Hu, Johnny H; Seidman, Christine E

2018-01-24

Human induced pluripotent stem cells (hiPSCs) can be used to mass produce surrogates of human tissues, enabling new advances in drug screening, disease modeling, and cell therapy. Recent developments in clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing technology use homology-directed repair (HDR) to efficiently generate custom hiPSC lines harboring a variety of genomic insertions and deletions. Thus, hiPSCs that encode an endogenous protein fused to a fluorescent reporter protein can be rapidly created by employing CRISPR/Cas9 genome editing, enhancing HDR efficiency and optimizing homology arm length. These fluorescently tagged hiPSCs can be used to visualize protein function and dynamics in real time as cells proliferate and differentiate. Given that nearly any intracellular protein can be fluorescently tagged, this system serves as a powerful tool to facilitate new discoveries across many biological disciplines. In this unit, we present protocols for the design, generation, and monoclonal expansion of genetically customized hiPSCs encoding fluorescently tagged endogenous proteins. © 2018 by John Wiley & Sons, Inc. Copyright © 2018 John Wiley & Sons, Inc.

Photo-triggered fluorescent theranostic prodrugs as DNA alkylating agents for mechlorethamine release and spatiotemporal monitoring.

PubMed

Cao, Yanting; Pan, Rong; Xuan, Weimin; Wei, Yongyi; Liu, Kejian; Zhou, Jiahong; Wang, Wei

2015-06-28

We describe a new theranostic strategy for selective delivery and spatiotemporal monitoring of mechlorethamine, a DNA alkylating agent. A photo-responsive prodrug is designed and composed of a photolabile o-nitrophenylethyl group, a DNA alkylating mechlorethamine drug and a coumarin fluorophore. Masking of the "N" in mechlorethamine in a positively charged state in the prodrug renders it inactive, non-toxic, selective and non-fluorescent. Indeed, the stable prodrug shows negligible cytotoxicity towards normal cells with and without UV activation and is completely non-fluorescent. However, upon photo-irradiation, the active mechlorethamine is released and induces efficient DNA cross-links, accompanied by a strong fluorescence enhancement (152 fold). Furthermore, DNA cross-linking activity from the release can be transformed into anticancer activity observed in in vitro studies of tumor cells. Importantly, the drug release progress and the movement can be conveniently monitored by fluorescence spectroscopy. The mechanistic study proves that the DNA cross-linking activity is mainly due to the release of DNA alkylating mechlorethamine. Altogether, the studies show the power of the theranostic strategy for efficient therapy in cancer treatment.

Fluorescent Lamp Replacement Study

DTIC Science & Technology

2017-07-01

friendly products, advances in efficiency, and lower production costs for lamps. The conversion of fluorescent bulbs to LED technology has many benefits ...of 4727 W. An economic analysis was calculated to compare the various lighting technologies that were implemented at ATC and the cost benefits ...the various lighting technologies that were implemented at ATC and the cost benefits of each, a lifecycle comparison was made between the fluorescent

Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging.

PubMed

Chen, Bin; Feng, Guangxue; He, Bairong; Goh, Chiching; Xu, Shidang; Ramos-Ortiz, Gabriel; Aparicio-Ixta, Laura; Zhou, Jian; Ng, Laiguan; Zhao, Zujin; Liu, Bin; Tang, Ben Zhong

2016-02-10

Robust luminescent dyes with efficient two-photon fluorescence are highly desirable for biological imaging applications, but those suitable for organic dots fabrication are still rare because of aggregation-caused quenching. In this work, a red fluorescent silole, 2,5-bis[5-(dimesitylboranyl)thiophen-2-yl]-1-methyl-1,3,4-triphenylsilole ((MesB)2 DTTPS), is synthesized and characterized. (MesB)2 DTTPS exhibits enhanced fluorescence efficiency in nanoaggregates, indicative of aggregation-enhanced emission (AEE). The organic dots fabricated by encapsulating (MesB)2 DTTPS within lipid-PEG show red fluorescence peaking at 598 nm and a high fluorescence quantum yield of 32%. Upon excitation at 820 nm, the dots show a large two-photon absorption cross section of 3.43 × 10(5) GM, which yields a two-photon action cross section of 1.09 × 10(5) GM. These (MesB)2 DTTPS dots show good biocompatibility and are successfully applied to one-photon and two-photon fluorescence imaging of MCF-7 cells and two-photon in vivo visualization of the blood vascular of mouse muscle in a high-contrast and noninvasive manner. Moreover, the 3D blood vasculature located at the mouse ear skin with a depth of over 100 μm can also be visualized clearly, providing the spatiotemporal information about the whole blood vascular network. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Turn-off fluorescence sensor for the detection of ferric ion in water using green synthesized N-doped carbon dots and its bio-imaging.

PubMed

Edison, Thomas Nesakumar Jebakumar Immanuel; Atchudan, Raji; Shim, Jae-Jin; Kalimuthu, Senthilkumar; Ahn, Byeong-Cheol; Lee, Yong Rok

2016-05-01

This paper reports turn-off fluorescence sensor for Fe(3+) ion in water using fluorescent N-doped carbon dots as a probe. A simple and efficient hydrothermal carbonization of Prunus avium fruit extract for the synthesis of fluorescent nitrogen-doped carbon dots (N-CDs) is described. This green approach proceeds quickly and provides good quality N-CDs. The mean size of synthesized N-CDs was approximately 7nm calculated from the high-resolution transmission electron microscopic images. X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy revealed the presence of -OH, -NH2, -COOH, and -CO functional groups over the surface of CDs. The N-CDs showed excellent fluorescent properties, and emitted blue fluorescence at 411nm upon excitation at 310nm. The calculated quantum yield of the synthesized N-CDs is 13% against quinine sulfate as a reference fluorophore. The synthesized N-CDs were used as a fluorescent probe towards the selective and sensitive detection of biologically important Fe(3+) ions in water by fluorescence spectroscopy and for bio-imaging of MDA-MB-231 cells. The limit of detection (LOD) and the Stern-Volmer quenching constant for the synthesized N-CDs were 0.96μM and 2.0958×10(3)M of Fe(3+) ions. The green synthesized N-CDs are efficiently used as a promising candidate for the detection of Fe(3+) ions and bio-imaging. Copyright © 2016 Elsevier B.V. All rights reserved.

Size-dependent fluorescence of bioaerosols: Mathematical model using fluorescing and absorbing molecules in bacteria

DOE PAGES

Hill, Steven C.; Williamson, Chatt C.; Doughty, David C.; ...

2015-02-02

This paper uses a mathematical model of fluorescent biological particles composed of bacteria and/or proteins (mostly as in Hill et al., 2013 [23]) to investigate the size-dependence of the total fluorescence emitted in all directions. The model applies to particles which have negligible reabsorption of fluorescence within the particle. The specific particles modeled here are composed of ovalbumin and of a generic Bacillus. The particles need not be spherical, and in some cases need not be homogeneous. However, the results calculated in this paper are for spherical homogeneous particles. Light absorbing and fluorescing molecules included in the model are aminomore » acids, nucleic acids, and several coenzymes. Here the excitation wavelength is 266 nm. The emission range, 300 to 370 nm, encompasses the fluorescence of tryptophan. The fluorescence cross section (C F) is calculated and compared with one set of published measured values. We investigate power law (Ad y) approximations to C F, where d is diameter, and A and y are parameters adjusted to fit the data, and examine how y varies with d and composition, including the fraction as water. The particle's fluorescence efficiency (Q F=C F/geometric-cross-section) can be written for homogeneous particles as Q absR F, where Q abs is the absorption efficiency, and R F, the fraction of the absorbed light emitted as fluorescence, is independent of size and shape. When Q F is plotted vs. m id or mi(m r-1)d, where m=m r+im i is the complex refractive index, the plots for different fractions of water in the particle tend to overlap.« less

Studies on Cation-induced Thylakoid Membrane Stacking, Fluorescence Yield, and Photochemical Efficiency 1

PubMed Central

Jennings, Robert Charles; Forti, Giorgio; Gerola, Paolo Domenico; Garlaschi, Flavio Massimo

1978-01-01

Trypsin digestion of photosynthetic membranes isolated from spinach (Spinacia oleracea L.) leaves eliminates the cation stimulation of chlorophyll fluorescence. High concentrations of cations protect the fluorescence yield against trypsin digestion, and the cation specificity for this protection closely resembles that required for the stimulation of fluorescence by cations. Trypsin digestion reverses cation-induced thylakoid stacking, and the time course of this effect seems to parallel that of the reversal of cation fluorescence. High concentrations of cations protect thylakoid stacking and cation-stimulated fluorescence alike. The cation stimulation of photosytem II photochemistry remains intact after trypsinization has reversed both cation-induced thylakoid stacking and fluorescence yield. It is concluded that cation-stimulated fluorescence yield, and not the cation stimulation of photosystem II photochemistry, is associated with thylakoid membrane stacking. ImagesFig. 2Fig. 3 PMID:16660630

Fluorescent nanoscale zinc(II)-carboxylate coordination polymers for explosive sensing.

PubMed

Zhang, Chengyi; Che, Yanke; Zhang, Zengxing; Yang, Xiaomei; Zang, Ling

2011-02-28

Fluorescent nanoscale coordination polymers with cubic morphology and long range ordered structure were fabricated and exhibited efficient sensing for both nitroaromatic explosive and nitromethane due to large surface area to volume ratio and strong binding affinity to explosive molecules.

Fiber optic biofluorometer for physiological research on muscle slices

NASA Astrophysics Data System (ADS)

Belz, Mathias; Dendorfer, Andreas; Werner, Jan; Lambertz, Daniel; Klein, Karl-Friedrich

2016-03-01

A focus of research in cell physiology is the detection of Ca2+, NADH, FAD, ATPase activity or membrane potential, only to name a few, in muscle tissues. In this work, we report on a biofluorometer using ultraviolet light emitting diodes (UV-LEDs), optical fibers and two photomultipliers (PMTs) using synchronized fluorescence detection with integrated background correction to detect free calcium, Ca2+, in cardiac muscle tissue placed in a horizontal tissue bath and a microscope setup. Fiber optic probes with imaging optics have been designed to transport excitation light from the biofluorometer's light output to a horizontal tissue bath and to collect emission light from a tissue sample of interest to two PMTs allowing either single excitation / single emission or ratiometric, dual excitation / single emission or single excitation / dual emission fluorescence detection of indicator dyes or natural fluorophores. The efficient transport of light from the excitation LEDs to the tissue sample, bleaching effects of the excitation light in both, polymer and fused silica-based fibers will be discussed. Furthermore, a new approach to maximize light collection of the emission light using high NA fibers and high NA coupling optics will be shown. Finally, first results on Ca2+ measurements in cardiac muscle slices in a traditional microscope setup and a horizontal tissue bath using fiber optic probes will be introduced and discussed.

Inkjet printed fluorescent nanorod layers exhibit superior optical performance over quantum dots

NASA Astrophysics Data System (ADS)

Halivni, Shira; Shemesh, Shay; Waiskopf, Nir; Vinetsky, Yelena; Magdassi, Shlomo; Banin, Uri

2015-11-01

Semiconductor nanocrystals exhibit unique fluorescence properties which are tunable in size, shape and composition. The high quantum yield and enhanced stability have led to their use in biomedical imaging and flat panel displays. Here, semiconductor nanorod based inkjet inks are presented, overcoming limitations of the commonly reported quantum dots in printing applications. Fluorescent seeded nanorods were found to be outstanding candidates for fluorescent inks, due to their low particle-particle interactions and negligible self-absorption. This is manifested by insignificant emission shifts upon printing, even in highly concentrated printed layers and by maintenance of a high fluorescence quantum yield, unlike quantum dots which exhibit fluorescence wavelength shifts and quenching effects. This behavior results from the reduced absorption/emission overlap, accompanied by low energy transfer efficiencies between the nanorods as supported by steady state and time resolved fluorescence measurements. The new seeded nanorod inks enable patterning of thin fluorescent layers, for demanding light emission applications such as signage and displays.Semiconductor nanocrystals exhibit unique fluorescence properties which are tunable in size, shape and composition. The high quantum yield and enhanced stability have led to their use in biomedical imaging and flat panel displays. Here, semiconductor nanorod based inkjet inks are presented, overcoming limitations of the commonly reported quantum dots in printing applications. Fluorescent seeded nanorods were found to be outstanding candidates for fluorescent inks, due to their low particle-particle interactions and negligible self-absorption. This is manifested by insignificant emission shifts upon printing, even in highly concentrated printed layers and by maintenance of a high fluorescence quantum yield, unlike quantum dots which exhibit fluorescence wavelength shifts and quenching effects. This behavior results from the reduced absorption/emission overlap, accompanied by low energy transfer efficiencies between the nanorods as supported by steady state and time resolved fluorescence measurements. The new seeded nanorod inks enable patterning of thin fluorescent layers, for demanding light emission applications such as signage and displays. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr06248a

A ratiometric fluorescent quantum dots based biosensor for organophosphorus pesticides detection by inner-filter effect.

PubMed

Yan, Xu; Li, Hongxia; Han, Xiaosong; Su, Xingguang

2015-12-15

In this work, we develop a novel and sensitive sensor for the detection of organophosphorus pesticides based on the inner-filter effect (IFE) between gold nanoparticles (AuNPs) and ratiometric fluorescent quantum dots (RF-QDs). The RF-QDs has been designed by hybridizing two differently colored CdTe QDs, in which the red emissive QDs entrapped in the silica sphere acting as the reference signal, and the green emissive QDs covalently attached on the silica surface serving as the response signal.The fluorescence of RF-QDs could be quenched by AuNPs based on IFE. Protamine could effectively turn on the fluorescence due to the electrostatic attraction between protamine and AuNPs. Trypsin can easily hydrolyze protamine, leading to the quench of the fluorescence. Then, the fluorescence could be recovered again by the addition of parathion-methyl (PM) which could inhibit the activity of trypsin. By measuring the fluorescence of RF-QDs, the inhibition efficiency of PM to trypsin activity was evaluated. Under the optimized conditions, the inhibition efficiency was proportional to the logarithm of PM concentration in the range of 0.04-400 ng mL(-1), with a detection limit of 0.018 ng mL(-1). Furthermore, the simple and convenient method had been used for PM detection in environmental and agricultural samples with satisfactory results. Copyright © 2015 Elsevier B.V. All rights reserved.

Subnanosecond fluorescence spectroscopy of human serum albumin as a method to estimate the efficiency of the depression therapy

NASA Astrophysics Data System (ADS)

Syrejshchikova, T. I.; Gryzunov, Yu. A.; Smolina, N. V.; Komar, A. A.; Uzbekov, M. G.; Misionzhnik, E. J.; Maksimova, N. M.

2010-05-01

The efficiency of the therapy of psychiatric diseases is estimated using the fluorescence measurements of the conformational changes of human serum albumin in the course of medical treatment. The fluorescence decay curves of the CAPIDAN probe (N-carboxyphenylimide of the dimethylaminonaphthalic acid) in the blood serum are measured. The probe is specifically bound to the albumin drug binding sites and exhibits fluorescence as a reporter ligand. A variation in the conformation of the albumin molecule substantially affects the CAPIDAN fluorescence decay curve on the subnanosecond time scale. A subnanosecond pulsed laser or a Pico-Quant LED excitation source and a fast photon detector with a time resolution of about 50 ps are used for the kinetic measurements. The blood sera of ten patients suffering from depression and treated at the Institute of Psychiatry were preliminary clinically tested. Blood for analysis was taken from each patient prior to the treatment and on the third week of treatment. For ten patients, the analysis of the fluorescence decay curves of the probe in the blood serum using the three-exponential fitting shows that the difference between the amplitudes of the decay function corresponding to the long-lived (9 ns) fluorescence of the probe prior to and after the therapeutic procedure reliably differs from zero at a significance level of 1% ( p < 0.01).

Visualizing double-stranded RNA distribution and dynamics in living cells by dsRNA binding-dependent fluorescence complementation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cheng, Xiaofei; College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, Zhejiang 310036; Deng, Ping

Double-stranded RNA (dsRNA) is an important type of RNA that plays essential roles in diverse cellular processes in eukaryotic organisms and a hallmark in infections by positive-sense RNA viruses. Currently, no in vivo technology has been developed for visualizing dsRNA in living cells. Here, we report a dsRNA binding-dependent fluorescence complementation (dRBFC) assay that can be used to efficiently monitor dsRNA distribution and dynamics in vivo. The system consists of two dsRNA-binding proteins, which are fused to the N- and C-terminal halves of the yellow fluorescent protein (YFP). Binding of the two fusion proteins to a common dsRNA brings themore » split YFP halves in close proximity, leading to the reconstitution of the fluorescence-competent structure and restoration of fluorescence. Using this technique, we were able to visualize the distribution and trafficking of the replicative RNA intermediates of positive-sense RNA viruses in living cells. - Highlights: • A live-cell imaging system was developed for visualizing dsRNA in vivo. • It uses dsRNA binding proteins fused with two halves of a fluorescent protein. • Binding to a common dsRNA enables the reporter to become fluorescent. • The system can efficiently monitor viral RNA replication in living cells.« less

Extended Fluorescent Resonant Energy Transfer in DNA Constructs

NASA Astrophysics Data System (ADS)

Oh, Taeseok

This study investigates the use of surfactants and metal cations for the enhancement of long range fluorescent resonant energy transfer (FRET) and the antenna effect in DNA structures with multiple fluorescent dyes. Double-stranded (ds) DNA structures were formed by hybridization of 21mer DNA oligonucleotides with different arrangements of three fluorescent TAMRA donor dyes with two different complementary 21mer oligonucleotides with one fluorescent TexasRed acceptor dye. In such DNA structures, hydrophobic interactions between the fluorescent dyes in close proximity produces dimerization which along with other quenching mechanisms leads to significant reduction of fluorescent emission properties. Addition of the surfactants Triton X-100, cetyltrimethyl ammonium bromide (CTAB) and sodium dodecyl sulfate (SDS) along with sodium cations (Na+) and divalent magnesium cations (Mg 2+) were tested for their ability to reduce quenching of the fluorescent dyes and improve overall fluorescent emission, the long range FRET and the antenna effect properties. When the neutral (uncharged) surfactant Triton X-100 was added to the FRET ds-DNA hybrid structures with three TAMRA donors and one TexasRed acceptor, dye dimerization and emission quenching remained unaffected. However, for the positively charged CTAB surfactant at concentrations of 100 uM or higher, the neutralization of the negatively charged ds-DNA backbone by the cationic surfactant micelles was found to reduce TAMRA dye dimerization and emission quenching and improve TexasRed quantum yield, resulting in much higher FRET efficiencies and an enhanced antenna effect. This improvement is likely due to the CTAB molecules covering or sheathing the fluorescent donor and acceptor dyes which breaks up the dimerized dye complexes and prevents further quenching from interactions with water molecules and guanine bases in the DNA structure. While the negatively charged SDS surfactant alone was not able to reduce dimerization and emission quenching due to repulsive forces between DNA and SDS micelles, the addition of cations such as sodium ions (Na+) and divalent magnesium ions (Mg2+) did lead to a significant reduction in the dimerization and emission quenching resulting in much higher FRET efficiency and an enhanced antenna effect. It appears that when the repulsive electrostatic forces are screened by the cations (Mg2+ in particular), the SDS micelles can approach the FRET ds-DNA structures thereby sheathing or insulating the TAMRA and TexasRed dyes. Overall, the study provides a viable strategy for using combinations of surfactants and cations to reduce adverse fluorescent dye and other quenching mechanisms and improve the overall long distance FRET efficiency and the antenna effect in DNA structures with multi-donor and single acceptor fluorescent dye groups.

Coral Pigments: Quantification Using HPLC and Detection by Remote Sensing

NASA Technical Reports Server (NTRS)

Cottone, Mary C.

1995-01-01

Widespread coral bleaching (loss of pigments of symbiotic dinoflagellates), and the corresponding decline in coral reef health worldwide, mandates the monitoring of coral pigmentation. Samples of the corals Porites compressa and P. lobata were collected from a healthy reef at Puako, Hawaii, and chlorophyll (chl) a, peridinin, and Beta-carotene (Beta-car) were quantified using reverse-phase high performance liquid chromatography (HPLC). Detailed procedures are presented for the extraction of the coral pigments in 90% acetone, and the separation, identification, and quantification of the major zooxanthellar pigments using spectrophotometry and a modification of the HPLC system described by Mantoura and Llewellyn (1983). Beta-apo-8-carotenal was found to be inadequate as in internal standard, due to coelution with chl b and/or chl a allomer in the sample extracts. Improvements are suggested, which may result in better resolution of the major pigments and greater accuracy in quantification. Average concentrations of peridinin, chl a, and Beta-car in corals on the reef were 5.01, 8.59, and 0.29, micro-grams/cm(exp 2), respectively. Average concentrations of peridinin and Beta-car did not differ significantly between the two coral species sampled; however, the mean chl a concentration in P. compressa specimens (7.81 ,micro-grams/cm(exp 2) was significantly lower than that in P. lobata specimens (9.96 11g/cm2). Chl a concentrations determined spectrophotometrically were significantly higher than those generated through HPLC, suggesting that spectrophotometry overestimates chl a concentrations. The average ratio of chl a-to-peridinin concentrations was 1.90, with a large (53%) coefficient of variation and a significant difference between the two species sampled. Additional data are needed before conclusions can be drawn regarding average pigment concentrations in healthy corals and the consistency of the chl a/peridinin ratio. The HPLC pigment concentration values contribute to the limited database of pigment concentrations in healthy corals, from which quantitative definitions of 'healthy' vs. 'bleached' coral may emerge. They also serve as ground-truth, corresponding to fluorescence data collected from the reef at Puako using airborne remote sensing of laser induced fluorescence. Fluorescence spectra from several overflights using the NASA AOL (airborne oceanographic lidar) system show consistent chlorphyll fluorescence peaks around 685 nm, as well as consistence peaks in the 400-600 nm range which may emanate from granules in the coral tissue. These data, along with results from previous studies of coral fluorescence, suggest that remote sensing of laser-induced fluorescence may become a rapid and efficient means of monitoring coral pigmentation and coral reef bleaching.

Tomato seeds maturity detection system based on chlorophyll fluorescence

NASA Astrophysics Data System (ADS)

Li, Cuiling; Wang, Xiu; Meng, Zhijun

2016-10-01

Chlorophyll fluorescence intensity can be used as seed maturity and quality evaluation indicator. Chlorophyll fluorescence intensity of seed coats is tested to judge the level of chlorophyll content in seeds, and further to judge the maturity and quality of seeds. This research developed a detection system of tomato seeds maturity based on chlorophyll fluorescence spectrum technology, the system included an excitation light source unit, a fluorescent signal acquisition unit and a data processing unit. The excitation light source unit consisted of two high power LEDs, two radiators and two constant current power supplies, and it was designed to excite chlorophyll fluorescence of tomato seeds. The fluorescent signal acquisition unit was made up of a fluorescence spectrometer, an optical fiber, an optical fiber scaffolds and a narrowband filter. The data processing unit mainly included a computer. Tomato fruits of green ripe stage, discoloration stage, firm ripe stage and full ripe stage were harvested, and their seeds were collected directly. In this research, the developed tomato seeds maturity testing system was used to collect fluorescence spectrums of tomato seeds of different maturities. Principal component analysis (PCA) method was utilized to reduce the dimension of spectral data and extract principal components, and PCA was combined with linear discriminant analysis (LDA) to establish discriminant model of tomato seeds maturity, the discriminant accuracy was greater than 90%. Research results show that using chlorophyll fluorescence spectrum technology is feasible for seeds maturity detection, and the developed tomato seeds maturity testing system has high detection accuracy.

Computational efficient segmentation of cell nuclei in 2D and 3D fluorescent micrographs

NASA Astrophysics Data System (ADS)

De Vylder, Jonas; Philips, Wilfried

2011-02-01

This paper proposes a new segmentation technique developed for the segmentation of cell nuclei in both 2D and 3D fluorescent micrographs. The proposed method can deal with both blurred edges as with touching nuclei. Using a dual scan line algorithm its both memory as computational efficient, making it interesting for the analysis of images coming from high throughput systems or the analysis of 3D microscopic images. Experiments show good results, i.e. recall of over 0.98.

Remanagement of Singlet and Triplet Excitons in Single-Emissive-Layer Hybrid White Organic Light-Emitting Devices Using Thermally Activated Delayed Fluorescent Blue Exciplex.

PubMed

Liu, Xiao-Ke; Chen, Zhan; Qing, Jian; Zhang, Wen-Jun; Wu, Bo; Tam, Hoi Lam; Zhu, Furong; Zhang, Xiao-Hong; Lee, Chun-Sing

2015-11-25

A high-performance hybrid white organic light-emitting device (WOLED) is demonstrated based on an efficient novel thermally activated delayed fluorescence (TADF) blue exciplex system. This device shows a low turn-on voltage of 2.5 V and maximum forward-viewing external quantum efficiency of 25.5%, which opens a new avenue for achieving high-performance hybrid WOLEDs with simple structures. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Real-time monitoring of river water quality using in-line continuous acquisition of fluorescence excitation and emission matrices

NASA Astrophysics Data System (ADS)

Carstea, E.; Baker, A.; Johnson, R.; Reynolds, D. M.

2009-12-01

In-line fluorescence EEM monitoring has been performed over an eleven-day period for Bournbrook River, Birmingham, UK. River water was diverted to a portable laboratory via a continuous flow pump and filter system. Fluorescence excitation-emission matrices data was recorded every 3 minutes using a flow cell (1cm pathlength) coupled to a fiber optic probe. This real-time fluorescence EEM data (Excitation, 225-400 nm at 5 nm steps, emission, 280-500 nm at 2 nm steps) was collected 'in-line'and directly compared with the spectrophotometric properties and physical and chemical parameters of river water samples collected off-line at known time intervals. Over the monitoring period, minor pollution pulses from cross connections were detected and identified hourly along with a random diesel pollution event. This work addresses the practicalities of measuring and detecting fluorescence EEM in the field and discusses the potential of this technological approach for further understanding important hydrological and biogeochemical processes. Problems associated with fouling and system failure are also reported. Example of the data generated from the continuous fluorescence EEM monitoring.

Blood interference in fiber-optical based fluorescence guided resection of glioma using 5-aminolevulinic acid

NASA Astrophysics Data System (ADS)

Haj-Hosseini, Neda; Lowndes, Shannely; Salerud, Göran; Wårdell, Karin

2011-03-01

Fluorescence guidance in brain tumor resection is performed intra-operatively where bleeding is included. When using fiber-optical probes, the transmission of light to and from the tissue is totally or partially blocked if a small amount of blood appears in front of the probe. Sometimes even after rinsing with saline, the remnant blood cells on the optical probe head, disturb the measurements. In such a case, the corresponding spectrum cannot be reliably quantified and is therefore discarded. The optimal case would be to calculate and take out the blood effect systematically from the collected signals. However, the first step is to study the pattern of blood interference in the fluorescence spectrum. In this study, a fiber-optical based fluorescence spectroscopy system with a laser excitation light of 405 nm (1.4 J/cm2) was used during fluorescence guided brain tumor resection using 5-aminolevulinic acid (5-ALA). The blood interference pattern in the fluorescence spectrum collected from the brain was studied in two patients. The operation situation was modeled in the laboratory by placing blood drops from the finger tip on the skin of forearm and the data was compared to the brain in vivo measurements. Additionally, a theoretical model was developed to simulate the blood interference pattern on the skin autofluorescence. The blood affects the collected fluorescence intensity and leaves traces of oxy and deoxy-hemoglobin absorption peaks. According to the developed theoretical model, the autofluorescence signal is considered to be totally blocked by an approximately 500 μm thick blood layer.

Development of real-time monitors for gaseous formaldehyde. Final report, 1 December 1988-30 September 1989

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kelly, T.J.; Barnes, R.H.

1990-11-01

Two new methods for real-time measurement of gaseous formaldehyde have been developed. One is a spectroscopic method based on direct fluorescence detection of gaseous formaldehyde following excitation with UV light. This method has been developed to the prototype stage by modifications of a commercial fluorescence SO2 detector to convert it to formaldehyde detection. The prototype spectroscopic formaldehyde monitor exhibits a detection limit of <30 ppbv, with a time response of about one minute. The second method is based on derivatization of formaldehyde in aqueous solution to form a fluorescent product. The detection of fluorescent product was made more sensitive bymore » using intense 254 nm light from a mercury lamp for excitation, thereby allowing use of a simple and efficient glass coil scrubber for collection of gaseous formaldehyde. The wet chemical formaldehyde monitor incorportating these improvements exhibits a detection limit for gaseous formaldehyde of 0.2 ppbv and for aqueous formaldehyde of 0.2 micromolar with time response of about one minute, following a lag time of 2 minutes. Both instruments were tested in the laboratory with gaseous formaldehyde standards, and the aqueous scrubbing/analysis method was field tested by continuous operation over a 10-day period in which outdoor and indoor air were sampled for alternate half-hour periods. A comparison of real-time (aqueous scrubbing/analysis) and integrated measurements, using dinitrophenylhydrazine (DNPH) impingers, showed close agreement between the real-time and DNPH data, even at concentrations as low as 1 ppbv.« less

Quenching of p-Cyanophenylalanine Fluorescence by Various Anions.

PubMed

Pazos, Ileana M; Roesch, Rachel M; Gai, Feng

2013-03-20

To expand the spectroscopic utility of the non-natural amino acid p -cyanophenylalanine (Phe CN ), we examine the quenching efficiencies of a series of commonly encountered anions toward its fluorescence. We find that iodide exhibits an unusually large Stern-Volmer quenching constant, making it a convenient choice in Phe CN fluorescence quenching studies. Indeed, using the villin headpiece subdomain as a testbed we demonstrate that iodide quenching of Phe CN fluorescence offers a convenient means to reveal protein conformational heterogeneity. Furthermore, we show that the amino group of Phe CN strongly quenches its fluorescence, suggesting that Phe CN could be used as a local pH sensor.

Monitoring the Behavior of Emerging Contaminants in Wastewater-Impacted Rivers Based on the Use of Fluorescence Excitation Emission Matrixes (EEM).

PubMed

Sgroi, Massimiliano; Roccaro, Paolo; Korshin, Gregory V; Vagliasindi, Federico G A

2017-04-18

This study investigated the applicability of fluorescence indexes based on the interpretation of excitation emission matrices (EEMs) by PARAFAC analysis and by selecting fluorescence intensities at a priori defined excitation/emission pairs as surrogates for monitoring the behavior of emerging organic compounds (EOCs) in two catchment basins impacted by wastewater discharges. Relevant EOC and EEM data were obtained for a 90 km stretch of the Simeto River, the main river in Sicily, and the smaller San Leonardo River, which was investigated for a 17 km stretch. The use of fluorescence indexes developed by these two different approaches resulted in similar observations. Changes of the fluorescence indexes that correspond to a group of humic-like fluorescing species were determined to be highly correlated with the concentrations of recalcitrant contaminants such as sucralose, sulfamethoxazole and carbamazepine, which are typical wastewater markers in river water. Changes of the fluorescence indexes related to tyrosine-like substances were well correlated with the concentrations of ibuprofen and caffeine, anthropogenic indicators of untreated wastewater discharges. Chemical oxygen demand and dissolved organic carbon concentrations were correlated with humic-like fluorescence indexes. The observed correlations were site-specific and characterized by different regression parameters for every collection event. Caffeine and carbamazepine showed correlations with florescence indexes in the San Leonardo River and in the alluvial plain stretch of the Simeto River, whereas sucralose, sulfamethoxazole and ibuprofen have always been well correlated in all the investigated river stretches. However, when data of different collection events from river stretches where correlations were observed were combined, good linear correlations were obtained for data sets generated via the normalization of the measured concentrations by the average value for the corresponding collection event. These results show that fluorescence based indexes can be used to monitor the behavior of some trace organic contaminants in wastewater impacted rivers and to track wastewater discharges in streams and rivers.

Reversible intermolecular energy transfer between saturated amines and benzene in non-polar solution

NASA Astrophysics Data System (ADS)

Halpern, Arthur M.; Wryzykowska, Krystyna

1981-01-01

Excitation of a mixture of dimethylethylamine (DEMA) and benzene in n-hexane at 222 nm primarily produces excited amine, while at 261 nm excited benzene predominantly results. The fluorescence spectra appreciably overlap. With 222 nm excitation, DEMA fluorescence is quenched by benzene at the diffusion-controlled rate; this quenching results with nearly unit efficiency in sensitized benzene fluorescence. With 261 nm excitation, some sensitized DEMA fluorescence is observed: the rate constant for tins process is ≈ 2.6 × 10 9 M -1 s -1.

NIR-Mediated Nanohybrids of Upconversion Nanophosphors and Fluorescent Conjugated Polymers for High-Efficiency Antibacterial Performance Based on Fluorescence Resonance Energy Transfer.

PubMed

Li, Junting; Zhao, Qi; Shi, Feng; Liu, Chenghui; Tang, Yanli

2016-12-01

A novel nanohybrid comprised of upconversion nanophosphors (UCNPs) and fluorescent conjugated polymers (PFVCN) is rationally fabricated. The new UCNP/PFVCN nanohybrids combine the excellent antibacterial ability of PFVCN and the near IR (NIR) absorbing property of UCNPs, which allows for NIR-mediated antibacterial through the effective fluorescence resonance energy transfer from UCNPs to PFVCN accompanied with generation of reactive oxygen species to kill bacteria. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Understanding Solar Induced Fluorescence: Building up from Leaf Scale Measurements (Invited)

NASA Astrophysics Data System (ADS)

Berry, J. A.; Van der Tol, C.; Frankenberg, C.; Joiner, J.; Guanter, L.

2013-12-01

Measurements of chlorophyll fluorescence have long been a key method for probing the mechanisms of photosynthesis in laboratory studies. Recent advances in satellite spectroscopy have enabled retrieval of chlorophyll fluorescence from terrestrial ecosystems at a global scale. Analyses of these retrievals show promising potential as an indicator of photosynthetic rate and of its response to environmental stress. This talk will explore the mechanistic basis for interpreting and modeling of solar induced chlorophyll fluorescence ( SIF). SIF is essentially a leak of photons from photosynthetic membranes, and it is, therefore, related to the flux of photons absorbed by chlorophyll and to biochemical processes that regulate the processing of these photons in macromolecuar complexes associated with photosystem II. Thus: SIF = aPAR * φF, where aPAR is the flux of absorbed photosynthetically active radiation and φF, is the yield (light-use efficiency) of fluorescence. (For simplicity we will ignore the transport of fluorescence from its sources to the sensor for the moment). This expression for SIF is similar to a common expression for photosynthesis or gross primary productivity, GPP = aPAR * LUE, where LUE, is the light-use-efficiency for CO2 uptake. These equations can be combined and simplified to illustrate the relationship between SIF and GPP; GPP =SIF *LUE / φF. The extent to which GPP is proportional to SIF hinges on the stability of the ratio, LUE / φF, and it leads to the key question to be considered here. What is the relationship between the light-use-efficiency for photosynthesis and that for fluorescence? Satellite retrievals of SIF occur at mid-day, conditions where the capacity for CO2 fixation usually limits the rate of photosynthesis. Under this condition the rate of the photo-acts must be down-regulated to protect from photo-damage. This balancing the source with the sink is accomplished by opening non-photochemical trapping centers that compete with the photochemical reactions at PSII and with the emission of fluorescence. We used simultaneous measurements of CO2 exchange and PAM fluorescence under laboratory conditions to evaluate the dependence of the kinetic constant for non-photochemical quenching (Kn) on relative quantum yield (or light use efficiency) for photosynthesis under a range of conditions, and we integrated this with a conventional parameterization for photosynthetic biochemistry to simulate SIF and GPP in the SCOPE model. We show that the fluorescence parameterization is able to reproduce leaf scale measurements well and that fluorescence measurements place new and more stringent constraints on photosynthesis parameterizations. The model was then used to evaluate the potential for using retrievals of SIF to estimate GPP, the integrated Vcmax of the canopy, the presence of water stress, and biophysical properties such as leaf angle distribution and chlorophyll content. This study strongly supports the usefulness of SIF, and illustrates some of the interactions that must be taken into account in interpreting these measurements.

Fish with red fluorescent eyes forage more efficiently under dim, blue-green light conditions.

PubMed

Harant, Ulrike Katharina; Michiels, Nicolaas Karel

2017-04-20

Natural red fluorescence is particularly conspicuous in the eyes of some small, benthic, predatory fishes. Fluorescence also increases in relative efficiency with increasing depth, which has generated speculation about its possible function as a "light organ" to detect cryptic organisms under bluish light. Here we investigate whether foraging success is improved under ambient conditions that make red fluorescence stand out more, using the triplefin Tripterygion delaisi as a model system. We repeatedly presented 10 copepods to individual fish (n = 40) kept under a narrow blue-green spectrum and compared their performance with that under a broad spectrum with the same overall brightness. The experiment was repeated for two levels of brightness, a shaded one representing 0.4% of the light present at the surface and a heavily shaded one with about 0.01% of the surface brightness. Fish were 7% more successful at catching copepods under the narrow, fluorescence-friendly spectrum than under the broad spectrum. However, this effect was significant under the heavily shaded light treatment only. This outcome corroborates previous predictions that fluorescence may be an adaptation to blue-green, heavily shaded environments, which coincides with the opportunistic biology of this species that lives in the transition zone between exposed and heavily shaded microhabitats.

Highly fluorescent and superparamagnetic nanosystem for biomedical applications

NASA Astrophysics Data System (ADS)

Cabrera, Mariana P.; E Cabral Filho, Paulo; Silva, Camila M. C. M.; Oliveira, Rita M.; Geraldes, Carlos F. G. C.; Castro, M. Margarida C. A.; Costa, Benilde F. O.; Henriques, Marta S. C.; Paixão, José A.; Carvalho, Luiz B., Jr.; Santos, Beate S.; Hallwass, Fernando; Fontes, Adriana; Pereira, Giovannia A. L.

2017-07-01

This work reports on highly fluorescent and superparamagnetic bimodal nanoparticles (BNPs) obtained by a simple and efficient method as probes for fluorescence analysis and/or contrast agents for MRI. These promising BNPs with small dimensions (ca. 17 nm) consist of superparamagnetic iron oxide nanoparticles (SPIONs) covalently bound with CdTe quantum dots (ca. 3 nm). The chemical structure of the magnetic part of BNPs is predominantly magnetite, with minor goethite and maghemite contributions, as shown by Mössbauer spectroscopy, which is compatible with the x-ray diffraction data. Their size evaluation by different techniques showed that the SPION derivatization process, in order to produce the BNPs, does not lead to a large size increase. The BNPs saturation magnetization, when corrected for the organic content of the sample, is ca. 68 emu g-1, which is only slightly reduced relative to the bare nanoparticles. This indicates that the SPION surface functionalization does not change considerably the magnetic properties. The BNP aqueous suspensions presented stability, high fluorescence, high relaxivity ratio (r 2/r 1 equal to 25) and labeled efficiently HeLa cells as can be seen by fluorescence analysis. These BNP properties point to their applications as fluorescent probes as well as negative T 2-weighted MRI contrast agents. Moreover, their potential magnetic response could also be used for fast bioseparation applications.

Detection of trace tetracycline in fish via synchronous fluorescence quenching with carbon quantum dots coated with molecularly imprinted silica

NASA Astrophysics Data System (ADS)

Yang, Ji; Lin, Zheng-Zhong; Nur, A.-Zha; Lu, Yan; Wu, Ming-Hui; Zeng, Jun; Chen, Xiao-Mei; Huang, Zhi-Yong

2018-02-01

A novel fluorescence-based sensor combining synchronous fluorescence spectroscopy (SFS) with molecularly imprinted polymers (MIPs) was fabricated with reverse microemulsion method. Tetracycline (TC), (3-aminopropyl) triethoxysilane (APTES), tetraethyl orthosilicate (TEOS) and carbon quantum dots (CDs) were used as template, functional monomer, cross-linker and signal sources respectively in the probe preparation. A synchronous fluorescence emission (λem) at 355 nm was observed for the prepared MIP-coated CDs (MIP@CDs) particles when the wavelength interval (Δλ) was set as 70 nm, and the synchronous fluorescence intensity could be rapidly and efficiently quenched by TC based on inner filter effect (IFE). The quenching efficiencies of synchronous fluorescence intensity was linearly fitted with tetracycline (TC) concentrations ranging from 0.1 to 50 μmol L- 1 with a detection limit (DL) of 9 nmol L- 1 (3σ, n = 9). The MIP@CDs was used as a probe to detect TC in fish samples with the recoveries ranging from 98.4% to 103.1% and the relative standard deviation less than 6.0%. The results illustrated that the as-prepared MIP@CDs could be applied to the detection of trace TC in fish samples with rapidity, high sensitivity and accuracy.

Ultra-sensitive and selective Hg{sup 2+} detection based on fluorescent carbon dots

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Ruihua; Li, Haitao; Kong, Weiqian

2013-07-15

Graphical abstract: Fluorescent carbon dots were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from PEG and demonstrated to show high selectivity toward Hg2+ ions detection. - Highlights: • FCDs were synthesized by one-step sodium hydroxide-assisted reflux method from PEG. • The FCDs emit blue photoluminescence and have upconversion fluorescent property. • The FCDs show ultra-sensitive detective ability for Hg{sup 2+} ions. - Abstract: Fluorescent carbon dots (FCDs) were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from poly(ethylene glycol) (PEG). The obtained FCDs exhibit excellent water-solubility and high stability. Under the UV irradiation, the FCDs could emit bright bluemore » photoluminescence, and also they were found to show excellent up-conversion fluorescence. It was further demonstrated that such FCDs can serve as effective fluorescent sensing platform for Hg{sup 2+} ions detection with ultra-sensitivity and selectivity. The sensing system achieved a limit of detection as low as 1 fM, which is much lower than all the previous reported sensing systems for Hg{sup 2+} ions detection. This FCDs sensing system has been successfully applied for the analysis of Hg{sup 2+} ions in water samples from river, lake, and tap water, showing good practical feasibility.« less

Sample collection system for gel electrophoresis

DOEpatents

Olivares, Jose A.; Stark, Peter C.; Dunbar, John M.; Hill, Karen K.; Kuske, Cheryl R.; Roybal, Gustavo

2004-09-21

An automatic sample collection system for use with an electrophoretic slab gel system is presented. The collection system can be used with a slab gel have one or more lanes. A detector is used to detect particle bands on the slab gel within a detection zone. Such detectors may use a laser to excite fluorescently labeled particles. The fluorescent light emitted from the excited particles is transmitted to low-level light detection electronics. Upon the detection of a particle of interest within the detection zone, a syringe pump is activated, sending a stream of buffer solution across the lane of the slab gel. The buffer solution collects the sample of interest and carries it through a collection port into a sample collection vial.

Advanced lighting guidelines: 1993. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Eley, C.; Tolen, T.M.; Benya, J.R.

1993-12-31

The 1993 Advanced Lighting Guidelines document consists of twelve guidelines that provide an overview of specific lighting technologies and design application techniques utilizing energy-efficient lighting practice. Lighting Design Practice assesses energy-efficient lighting strategies, discusses lighting issues, and explains how to obtain quality lighting design and consulting services. Luminaires and Lighting Systems surveys luminaire equipment designed to take advantage of advanced technology lamp products and includes performance tables that allow for accurate estimation of luminaire light output and power input. The additional ten guidelines -- Computer-Aided Lighting Design, Energy-Efficient Fluorescent Ballasts, Full-Size Fluorescent Lamps, Compact Fluorescent Lamps, Tungsten-Halogen Lamps, Metal Halidemore » and HPS Lamps, Daylighting and Lumen Maintenance, Occupant Sensors, Time Scheduling Systems, and Retrofit Control Technologies -- each provide a product technology overview, discuss current products on the lighting equipment market, and provide application techniques. This document is intended for use by electric utility personnel involved in lighting programs, lighting designers, electrical engineers, architects, lighting manufacturers` representatives, and other lighting professionals.« less

Fluorescence properties and energy transfer study of Er3+/Nd3+ doped fluorophosphate glass pumped at 800 and 980 nm for mid-infrared laser applications

NASA Astrophysics Data System (ADS)

Tian, Ying; Xu, Rongrong; Hu, Lili; Zhang, Junjie

2012-04-01

The fluorescence properties of 2.7 μm emission as well as near infrared emissions in Er3+/Nd3+ doped fluorophosphate glasses are investigated under 800 and 980 nm excitation. The fluorescence dynamics and energy transfer processes between Er and Nd ions in different pumping schemes are reported. Three Judd-Ofelt intensity parameters, energy transfer microparameters, and efficiency have been determined using the Judd-Ofelt and Förster-Dexter theories. The calculated energy transfer efficiency of the Er3+:4I13/2 level to the Nd3+:4I15/2 level is as high as 83.91%. The results indicate that Nd3+ may be an efficient sensitizer for Er3+ to obtain mid-infrared emission and the more suitable pumping scheme of 2.7 μm laser applications for Er3+/Nd3+ doped fluorophosphate glass is 980 nm excitation.

Adapting smartphones for low-cost optical medical imaging

NASA Astrophysics Data System (ADS)

Pratavieira, Sebastião.; Vollet-Filho, José D.; Carbinatto, Fernanda M.; Blanco, Kate; Inada, Natalia M.; Bagnato, Vanderlei S.; Kurachi, Cristina

2015-06-01

Optical images have been used in several medical situations to improve diagnosis of lesions or to monitor treatments. However, most systems employ expensive scientific (CCD or CMOS) cameras and need computers to display and save the images, usually resulting in a high final cost for the system. Additionally, this sort of apparatus operation usually becomes more complex, requiring more and more specialized technical knowledge from the operator. Currently, the number of people using smartphone-like devices with built-in high quality cameras is increasing, which might allow using such devices as an efficient, lower cost, portable imaging system for medical applications. Thus, we aim to develop methods of adaptation of those devices to optical medical imaging techniques, such as fluorescence. Particularly, smartphones covers were adapted to connect a smartphone-like device to widefield fluorescence imaging systems. These systems were used to detect lesions in different tissues, such as cervix and mouth/throat mucosa, and to monitor ALA-induced protoporphyrin-IX formation for photodynamic treatment of Cervical Intraepithelial Neoplasia. This approach may contribute significantly to low-cost, portable and simple clinical optical imaging collection.

High sensitivity optical molecular imaging system

NASA Astrophysics Data System (ADS)

An, Yu; Yuan, Gao; Huang, Chao; Jiang, Shixin; Zhang, Peng; Wang, Kun; Tian, Jie

2018-02-01

Optical Molecular Imaging (OMI) has the advantages of high sensitivity, low cost and ease of use. By labeling the regions of interest with fluorescent or bioluminescence probes, OMI can noninvasively obtain the distribution of the probes in vivo, which play the key role in cancer research, pharmacokinetics and other biological studies. In preclinical and clinical application, the image depth, resolution and sensitivity are the key factors for researchers to use OMI. In this paper, we report a high sensitivity optical molecular imaging system developed by our group, which can improve the imaging depth in phantom to nearly 5cm, high resolution at 2cm depth, and high image sensitivity. To validate the performance of the system, special designed phantom experiments and weak light detection experiment were implemented. The results shows that cooperated with high performance electron-multiplying charge coupled device (EMCCD) camera, precision design of light path system and high efficient image techniques, our OMI system can simultaneously collect the light-emitted signals generated by fluorescence molecular imaging, bioluminescence imaging, Cherenkov luminance and other optical imaging modality, and observe the internal distribution of light-emitting agents fast and accurately.

Enhanced optical fiber fluorometer using a periodic perturbation in the fiber core

NASA Astrophysics Data System (ADS)

Chiniforooshan, Yasser; Bock, Wojtek J.; Ma, Jianjun

2013-10-01

Tracing of the specific chemicals and biological agents in a solution is becoming a vital interest in health, security and safety industries. Although a number of standard laboratory-based testing systems exists for detecting such targets, but the fast, real-time and on-site methods could be more efficient and cost-effective. One of the most common ways to detect a target in the solution is to use the fluorophore molecules which will be selectively attached to the targets and will emit or quench the fluorescence in presence of the target. The fiber-optic fluorometers are developed for inexpensive and portable detection. In this paper, we explain a novel multi-segment fiber structure which uses the periodic perturbation on the side-wall of a highly multi-mode fiber to enhance collecting the fluorescent light. This periodic perturbation is fabricated and optimized on the core of the fiber using a CO2 laser. The theoretical explanation to show the physical principle of the structure is followed by the experimental evidence of its functioning.

Multiplexed immunosensing and kinetics monitoring in nanofluidic devices with highly enhanced target capture efficiency

PubMed Central

Lin, Yii-Lih; Huang, Yen-Jun; Teerapanich, Pattamon; Leïchlé, Thierry

2016-01-01

Nanofluidic devices promise high reaction efficiency and fast kinetic responses due to the spatial constriction of transported biomolecules with confined molecular diffusion. However, parallel detection of multiple biomolecules, particularly proteins, in highly confined space remains challenging. This study integrates extended nanofluidics with embedded protein microarray to achieve multiplexed real-time biosensing and kinetics monitoring. Implementation of embedded standard-sized antibody microarray is attained by epoxy-silane surface modification and a room-temperature low-aspect-ratio bonding technique. An effective sample transport is achieved by electrokinetic pumping via electroosmotic flow. Through the nanoslit-based spatial confinement, the antigen-antibody binding reaction is enhanced with ∼100% efficiency and may be directly observed with fluorescence microscopy without the requirement of intermediate washing steps. The image-based data provide numerous spatially distributed reaction kinetic curves and are collectively modeled using a simple one-dimensional convection-reaction model. This study represents an integrated nanofluidic solution for real-time multiplexed immunosensing and kinetics monitoring, starting from device fabrication, protein immobilization, device bonding, sample transport, to data analysis at Péclet number less than 1. PMID:27375819

Characterization of marine macroalgae by fluorescence signatures

NASA Technical Reports Server (NTRS)

Topinka, J. A.; Bellows, W. Korjeff; Yentsch, C. S.

1990-01-01

The feasibility of distinguishing macroalgal classes by their fluorescence signatures was investigated using narrow-waveband light to excite groups of accessory pigments in brown, red, and green macroalgae and measuring fluorescence emission at 685 nm. Results obtained on 20 marine macroalgae field-collected samples showed that fluorescence excitation signatures were relatively uniform within phylogenetic classes but were substantially different for different classes. It is suggested that it may be possible to characterize the type and the abundance of subtidal macroalgae from low-flying aircraft using existing laser-induced fluorescence methodology.

Layer-by-layer films and colloidal dispersions of graphene oxide nanosheets for efficient control of the fluorescence and aggregation properties of the cationic dye acridine orange.

PubMed

Hansda, Chaitali; Chakraborty, Utsav; Hussain, Syed Arshad; Bhattacharjee, Debajyoti; Paul, Pabitra Kumar

2016-03-15

Chemically derived graphene oxide (GO) nanosheets have received great deal of interest for technological application such as optoelectronic and biosensors. Aqueous dispersions of GO become an efficient template to induce the association of cationic dye namely Acridine Orange (AO). Interactions of AO with colloidal GO was governed by both electrostatic and π-π stacking cooperative interactions. The type of dye aggregations was found to depend on the concentration of GO in the mixed ensemble. Spectroscopic calculations revealed the formation of both H and J-type dimers, but H-type aggregations were predominant. Preparation of layer-by-layer (LbL) electrostatic self-assembled films of AO and GO onto poly (allylamine hydrochloride) (PAH) coated quartz substrate is also reported in this article. UV-Vis absorption, steady state and time resolve fluorescence and Raman spectroscopic techniques have been employed to explore the detail photophysical properties of pure AO, AO/GO mixed solution and AO/GO LbL films. Scanning electron microscopy was also used for visual evidence of the synthesized nanodimensional GO sheets. The fluorescence quenching of AO in the presence of GO in aqueous solution was due to the interfacial photoinduced electron transfer (PET) from photoexcited AO to GO i.e. GO acts as an efficient quenching agent for the fluorescence emission of AO. The quenching is found to be static in nature. Raman spectroscopic results also confirmed the interaction of AO with GO and the electron transfer. The formation of AO/GO complex via very fast excited state electron transfer mechanism may be proposed as to prepare GO-based fluorescence sensor for biomolecular detection without direct labeling the biomolecules by fluorescent probe. Copyright © 2015 Elsevier B.V. All rights reserved.

Layer-by-layer films and colloidal dispersions of graphene oxide nanosheets for efficient control of the fluorescence and aggregation properties of the cationic dye acridine orange

NASA Astrophysics Data System (ADS)

Hansda, Chaitali; Chakraborty, Utsav; Hussain, Syed Arshad; Bhattacharjee, Debajyoti; Paul, Pabitra Kumar

2016-03-01

Chemically derived graphene oxide (GO) nanosheets have received great deal of interest for technological application such as optoelectronic and biosensors. Aqueous dispersions of GO become an efficient template to induce the association of cationic dye namely Acridine Orange (AO). Interactions of AO with colloidal GO was governed by both electrostatic and π-π stacking cooperative interactions. The type of dye aggregations was found to depend on the concentration of GO in the mixed ensemble. Spectroscopic calculations revealed the formation of both H and J-type dimers, but H-type aggregations were predominant. Preparation of layer-by-layer (LbL) electrostatic self-assembled films of AO and GO onto poly (allylamine hydrochloride) (PAH) coated quartz substrate is also reported in this article. UV-Vis absorption, steady state and time resolve fluorescence and Raman spectroscopic techniques have been employed to explore the detail photophysical properties of pure AO, AO/GO mixed solution and AO/GO LbL films. Scanning electron microscopy was also used for visual evidence of the synthesized nanodimensional GO sheets. The fluorescence quenching of AO in the presence of GO in aqueous solution was due to the interfacial photoinduced electron transfer (PET) from photoexcited AO to GO i.e. GO acts as an efficient quenching agent for the fluorescence emission of AO. The quenching is found to be static in nature. Raman spectroscopic results also confirmed the interaction of AO with GO and the electron transfer. The formation of AO/GO complex via very fast excited state electron transfer mechanism may be proposed as to prepare GO-based fluorescence sensor for biomolecular detection without direct labeling the biomolecules by fluorescent probe.

Quantitation of circulating tumor cells in blood samples from ovarian and prostate cancer patients using tumor-specific fluorescent ligands.

PubMed

He, Wei; Kularatne, Sumith A; Kalli, Kimberly R; Prendergast, Franklyn G; Amato, Robert J; Klee, George G; Hartmann, Lynn C; Low, Philip S

2008-10-15

Quantitation of circulating tumor cells (CTCs) can provide information on the stage of a malignancy, onset of disease progression and response to therapy. In an effort to more accurately quantitate CTCs, we have synthesized fluorescent conjugates of 2 high-affinity tumor-specific ligands (folate-AlexaFluor 488 and DUPA-FITC) that bind tumor cells >20-fold more efficiently than fluorescent antibodies. Here we determine whether these tumor-specific dyes can be exploited for quantitation of CTCs in peripheral blood samples from cancer patients. A CTC-enriched fraction was isolated from the peripheral blood of ovarian and prostate cancer patients by an optimized density gradient centrifugation protocol and labeled with the aforementioned fluorescent ligands. CTCs were then quantitated by flow cytometry. CTCs were detected in 18 of 20 ovarian cancer patients (mean 222 CTCs/ml; median 15 CTCs/ml; maximum 3,118 CTCs/ml), whereas CTC numbers in 16 gender-matched normal volunteers were negligible (mean 0.4 CTCs/ml; median 0.3 CTCs/ml; maximum 1.5 CTCs/ml; p < 0.001, chi(2)). CTCs were also detected in 10 of 13 prostate cancer patients (mean 26 CTCs/ml, median 14 CTCs/ml, maximum 94 CTCs/ml) but not in 18 gender-matched healthy donors (mean 0.8 CTCs/ml, median 1, maximum 3 CTC/ml; p < 0.0026, chi(2)). Tumor-specific fluorescent antibodies were much less efficient in quantitating CTCs because of their lower CTC labeling efficiency. Use of tumor-specific fluorescent ligands to label CTCs in peripheral blood can provide a simple, accurate and sensitive method for determining the number of cancer cells circulating in the bloodstream.

Rigidifying fluorescent linkers by metal-organic framework formation for fluorescence blue shift and quantum yield enhancement.

PubMed

Wei, Zhangwen; Gu, Zhi-Yuan; Arvapally, Ravi K; Chen, Ying-Pin; McDougald, Roy N; Ivy, Joshua F; Yakovenko, Andrey A; Feng, Dawei; Omary, Mohammad A; Zhou, Hong-Cai

2014-06-11

We demonstrate that rigidifying the structure of fluorescent linkers by structurally constraining them in metal-organic frameworks (MOFs) to control their conformation effectively tunes the fluorescence energy and enhances the quantum yield. Thus, a new tetraphenylethylene-based zirconium MOF exhibits a deep-blue fluorescent emission at 470 nm with a unity quantum yield (99.9 ± 0.5%) under Ar, representing ca. 3600 cm(-1) blue shift and doubled radiative decay efficiency vs the linker precursor. An anomalous increase in the fluorescence lifetime and relative intensity takes place upon heating the solid MOF from cryogenic to ambient temperatures. The origin of these unusual photoluminescence properties is attributed to twisted linker conformation, intramolecular hindrance, and framework rigidity.

Fluorescence Quenching of Alpha-Fetoprotein by Gold Nanoparticles: Effect of Dielectric Shell on Non-Radiative Decay

NASA Astrophysics Data System (ADS)

Zhu, Jian; Li, Jian-Jun; Wang, A.-Qing; Chen, Yu; Zhao, Jun-Wu

2010-09-01

Fluorescence quenching spectrometry was applied to study the interactions between gold colloidal nanoparticles and alpha-fetoprotein (AFP). Experimental results show that the gold nanoparticles can quench the fluorescence emission of adsorbed AFP effectively. Furthermore, the intensity of fluorescence emission peak decreases monotonously with the increasing gold nanoparticles content. A mechanism based on surface plasmon resonance-induced non-radiative decay was investigated to illuminate the effect of a dielectric shell on the fluorescence quenching ability of gold nanoparticles. The calculation results show that the increasing dielectric shell thickness may improve the monochromaticity of fluorescence quenching. However, high energy transfer efficiency can be obtained within a wide wavelength band by coating a thinner dielectric shell.

Sea Surface Scanner: An advanced catamaran to study the sea surface

NASA Astrophysics Data System (ADS)

Wurl, O.; Mustaffa, N. I. H.; Ribas Ribas, M.

2016-02-01

The Sea Surface Scanner is a remote-controlled catamaran with the capability to sample the sea-surface microlayer in high resolution. The catamaran is equipped with a suite of sensors to scan the sea surface on chemical, biological and physical parameters. Parameters include UV absorption, fluorescence spectra, chlorophyll-a, photosynthetic efficiency, chromophoric dissolved organic matter (CDOM), dissolved oxygen, pH, temperature, and salinity. A further feature is a capability to collect remotely discrete water samples for detailed lab analysis. We present the first high-resolution (< 30 sec) data on the sea surface microlayer. We discuss the variability of biochemical properties of the sea surface and its implication on air-sea interaction.

Mirrored pyramidal wells for simultaneous multiple vantage point microscopy.

PubMed

Seale, K T; Reiserer, R S; Markov, D A; Ges, I A; Wright, C; Janetopoulos, C; Wikswo, J P

2008-10-01

We report a novel method for obtaining simultaneous images from multiple vantage points of a microscopic specimen using size-matched microscopic mirrors created from anisotropically etched silicon. The resulting pyramidal wells enable bright-field and fluorescent side-view images, and when combined with z-sectioning, provide additional information for 3D reconstructions of the specimen. We have demonstrated the 3D localization and tracking over time of the centrosome of a live Dictyostelium discoideum. The simultaneous acquisition of images from multiple perspectives also provides a five-fold increase in the theoretical collection efficiency of emitted photons, a property which may be useful for low-light imaging modalities such as bioluminescence, or low abundance surface-marker labelling.

Intravital imaging of cardiac function at the single-cell level.

PubMed

Aguirre, Aaron D; Vinegoni, Claudio; Sebas, Matt; Weissleder, Ralph

2014-08-05

Knowledge of cardiomyocyte biology is limited by the lack of methods to interrogate single-cell physiology in vivo. Here we show that contracting myocytes can indeed be imaged with optical microscopy at high temporal and spatial resolution in the beating murine heart, allowing visualization of individual sarcomeres and measurement of the single cardiomyocyte contractile cycle. Collectively, this has been enabled by efficient tissue stabilization, a prospective real-time cardiac gating approach, an image processing algorithm for motion-artifact-free imaging throughout the cardiac cycle, and a fluorescent membrane staining protocol. Quantification of cardiomyocyte contractile function in vivo opens many possibilities for investigating myocardial disease and therapeutic intervention at the cellular level.

Laser Diagnostic Method for Plasma Sheath Potential Mapping

NASA Astrophysics Data System (ADS)

Walsh, Sean P.

Electric propulsion systems are gaining popularity in the aerospace field as a viable option for long term positioning and thrusting applications. In particular, Hall thrusters have shown promise as the primary propulsion engine for space probes during interplanetary journeys. However, the interaction between propellant xenon ions and the ceramic channel wall continues to remain a complex issue. The most significant source of power loss in Hall thrusters is due to electron and ion currents through the sheath to the channel wall. A sheath is a region of high electric field that separates a plasma from a wall or surface in contact. Plasma electrons with enough energy to penetrate the sheath may result emission of a secondary electron from the wall. With significant secondary electron emission (SEE), the sheath voltage is reduced and so too is the electron retarding electric field. Therefore, a lower sheath voltage further increases the particle loss to the wall of a Hall thruster and leads to plasma cooling and lower efficiency. To further understand sheath dynamics, laser-induced fluorescence is employed to provide a non-invasive, in situ, and spatially resolved technique for measuring xenon ion velocity. By scanning the laser wavelength over an electronic transition of singly ionized xenon and collecting the resulting fluorescence, one can determine the ion velocity from the Doppler shifted absorption. Knowing the velocity at multiple points in the sheath, it can be converted to a relative electric potential profile which can reveal a lot about the plasma-wall interaction and the severity of SEE. The challenge of adequately measuring sheath potential profiles is optimizing the experiment to maximize the signal-to-noise ratio. A strong signal with low noise, enables high resolution measurements and increases the depth of measurement in the sheath, where the signal strength is lowest. Many improvements were made to reduce the background luminosity, increase the fluorescence intensity and collection efficiency, and optimize the signal processing equipment. Doing so has allowed for a spatial resolution of 60 microns and a maximum depth of measurement of 2 mm depending on conditions. Sheaths surrounding common Hall thruster ceramics at various plasma conditions were measured in an attempt to determine the effect of SEE and a numerical analysis of the plasma-wall interactions was conducted to further understand the phenomena and compare against obtained data.

Use of light absorbers to alter optical interrogation with epi-illumination and transillumination in three-dimensional cardiac models

NASA Astrophysics Data System (ADS)

Ramshesh, Venkat K.; Knisley, Stephen B.

2006-03-01

Cardiac optical mapping currently provides 2-D maps of transmembrane voltage-sensitive fluorescence localized near the tissue surface. Methods for interrogation at different depths are required for studies of arrhythmias and the effects of defibrillation shocks in 3-D cardiac tissue. We model the effects of coloading with a dye that absorbs excitation or fluorescence light on the radius and depth of the interrogated region with specific illumination and collection techniques. Results indicate radii and depths of interrogation are larger for transillumination versus epi-illumination, an effect that is more pronounced for broad-field excitation versus laser scanner. Coloading with a fluorescence absorber lessens interrogated depth for epi-illumination and increases it for transillumination, which is confirmed with measurements using transillumination of heart tissue slices. Coloading with an absorber of excitation light consistently decreases the interrogated depths. Transillumination and coloading also decrease the intensities of collected fluorescence. Thus, localization can be modified with wavelength-specific absorbers at the expense of a reduction in fluorescence intensity.

A new non-resonant laser-induced fluorescence instrument for the airborne in situ measurement of formaldehyde

NASA Astrophysics Data System (ADS)

St. Clair, Jason M.; Swanson, Andrew K.; Bailey, Steven A.; Wolfe, Glenn M.; Marrero, Josette E.; Iraci, Laura T.; Hagopian, John G.; Hanisco, Thomas F.

2017-12-01

A new in situ instrument for gas-phase formaldehyde (HCHO), COmpact Formaldehyde FluorescencE Experiment (COFFEE), is presented. COFFEE utilizes non-resonant laser-induced fluorescence (NR-LIF) to measure HCHO, with 300 mW of 40 kHz 355 nm laser output exciting multiple HCHO absorption features. The resulting HCHO fluorescence is collected at 5 ns resolution, and the fluorescence time profile is fit to yield the ambient HCHO mixing ratio. Typical 1σ precision at ˜ 0 pptv HCHO is 150 pptv for 1 s data. The compact instrument was designed to operate with minimal in-flight operator interaction and infrequent maintenance (1-2 times per year). COFFEE fits in the wing pod of the Alpha Jet stationed at the NASA Ames Research Center and has successfully collected HCHO data on 27 flights through 2017 March. The frequent flights, combined with a potentially long-term data set, makes the Alpha Jet a promising platform for validation of satellite-based column HCHO.

A New Non-Resonant Laser-Induced Fluorescence Instrument for the Airborne in Situ Measurement of Formaldehyde

NASA Technical Reports Server (NTRS)

St. Clair, Jason M.; Swanson, Andrew K.; Bailey, Steven A.; Wolfe, Glenn M.; Marrero, Josette E.; Iraci, Laura T.; Hagopian, John G.; Hanisco, Thomas F.

2017-01-01

A new in situ instrument for gas-phase formaldehyde (HCHO), COmpact Formaldehyde FluorescencE Experiment (COFFEE), is presented. COFFEE utilizes nonresonant laser-induced fluorescence (NR-LIF) to measure HCHO, with 300 mW of 40 kHz 355 nm laser output exciting multiple HCHO absorption features. The resulting HCHO fluorescence is collected at 5 ns resolution, and the fluorescence time profile is fit to yield the ambient HCHO mixing ratio. Typical 1 sigma precision at approximately 0 pptv HCHO is 150 pptv for 1 s data. The compact instrument was designed to operate with minimal in-flight operator interaction and infrequent maintenance (1-2 times per year). COFFEE fits in the wing pod of the Alpha Jet stationed at the NASA Ames Research Center and has successfully collected HCHO data on 27 flights through 2017 March. The frequent flights, combined with a potentially long-term data set, makes the Alpha Jet a promising platform for validation of satellite-based column HCHO.

APTS and rGO co-functionalized pyrenated fluorescent nanonets for representative vapor phase nitroaromatic explosive detection

NASA Astrophysics Data System (ADS)

Guo, Linjuan; Zu, Baiyi; Yang, Zheng; Cao, Hongyu; Zheng, Xuefang; Dou, Xincun

2014-01-01

For the first time, flexible PVP/pyrene/APTS/rGO fluorescent nanonets were designed and synthesized via a one-step electrospinning method to detect representative subsaturated nitroaromatic explosive vapor. The functional fluorescent nanonets, which were highly stable in air, showed an 81% quenching efficiency towards TNT vapor (~10 ppb) with an exposure time of 540 s at room temperature. The nice performance of the nanonets was ascribed to the synergistic effects induced by the specific adsorption properties of APTS, the fast charge transfer properties and the effective π-π interaction with pyrene and TNT of rGO. Compared to the analogues of TNT, the PVP/pyrene/APTS/rGO nanonets showed notable selectivity towards TNT and DNT vapors. The explored functionalization method opens up brand new insight into sensitive and selective detection of vapor phase nitroaromatic explosives.For the first time, flexible PVP/pyrene/APTS/rGO fluorescent nanonets were designed and synthesized via a one-step electrospinning method to detect representative subsaturated nitroaromatic explosive vapor. The functional fluorescent nanonets, which were highly stable in air, showed an 81% quenching efficiency towards TNT vapor (~10 ppb) with an exposure time of 540 s at room temperature. The nice performance of the nanonets was ascribed to the synergistic effects induced by the specific adsorption properties of APTS, the fast charge transfer properties and the effective π-π interaction with pyrene and TNT of rGO. Compared to the analogues of TNT, the PVP/pyrene/APTS/rGO nanonets showed notable selectivity towards TNT and DNT vapors. The explored functionalization method opens up brand new insight into sensitive and selective detection of vapor phase nitroaromatic explosives. Electronic supplementary information (ESI) available: Vapor pressure of TNT and its analogues, fluorescence quenching kinetics, fluorescence quenching efficiencies and additional SEM images. See DOI: 10.1039/c3nr04960d

Aluminum nanostructures for ultraviolet plasmonics

NASA Astrophysics Data System (ADS)

Martin, Jérôme; Khlopin, Dmitry; Zhang, Feifei; Schuermans, Silvère; Proust, Julien; Maurer, Thomas; Gérard, Davy; Plain, Jérôme

2017-08-01

An electromagnetic field is able to produce a collective oscillation of free electrons at a metal surface. This allows light to be concentrated in volumes smaller than its wavelength. The resulting waves, called surface plasmons can be applied in various technological applications such as ultra-sensitive sensing, Surface Enhanced Raman Spectroscopy, or metal-enhanced fluorescence, to name a few. For several decades plasmonics has been almost exclusively studied in the visible region by using nanoparticles made of gold or silver as these noble metals support plasmonic resonances in the visible and near-infrared range. Nevertheless, emerging applications will require the extension of nano-plasmonics toward higher energies, in the ultraviolet range. Aluminum is one of the most appealing metal for pushing plasmonics up to ultraviolet energies. The subsequent applications in the field of nano-optics are various. This metal is therefore a highly promising material for commercial applications in the field of ultraviolet nano-optics. As a consequence, aluminum (or ultraviolet, UV) plasmonics has emerged quite recently. Aluminium plasmonics has been demonstrated efficient for numerous potential applications including non-linear optics, enhanced fluorescence, UV-Surface Enhanced Raman Spectroscopy, optoelectronics, plasmonic assisted solid-state lasing, photocatalysis, structural colors and data storage. In this article, different preparation methods developed in the laboratory to obtain aluminum nanostructures with different geometries are presented. Their optical and morphological characterizations of the nanostructures are given and some proof of principle applications such as fluorescence enhancement are discussed.

Investigation of Fluorescence Lifetime Quenching of Ru(bpy)[subscript 3][superscript 2+] by Oxygen Using a Pulsed Light-Emitting Diode

ERIC Educational Resources Information Center

Rusak, David A.; James, William H., III; Ferzola, Maria J.; Stefanski, Michael J.

2006-01-01

An experiment related to the measurement of fluorescence lifetime for an undergraduate instrumental analysis or physical chemistry laboratory that highlights relative rates of electronic transitions in molecules and introduces students to data collection of a pulsed signal is illustrated. The experiment of the long fluorescence lifetime of…

A NEW TRACER METHOD FOR ASSESSING HAND-TO-MOUTH TRANSFERS OF PESTICIDES

EPA Science Inventory

A common food chemical with high fluorescence intensity is being used in an evaluation of dermal transfer efficiencies as a surrogate for indoor pesticides chlorpyrifos and synthetic pyrethroids. The fluorescence monitoring system is based on an Olympus digital camera and the ...

Alstonine as a potential fluorescent marker for tiny tumor detection and imaging

NASA Astrophysics Data System (ADS)

Viallet, Pierre M.; Vo-Dinh, Tuan; Salmon, Jean-Marie; Watts, Wendi; Rocchi, Emmanuelle; Isola, Narayana R.; Rebillard, Xavier

1997-06-01

3,4,5,6,16,17-Hexadehydro-16-(methoxycarbolyl)-19(alpha) - methyl-20(alpha) -oxyohimbanium (alstonine) is a fluorescent alcaloid which is known to stain tumor cells more efficiently than normal. The interactions between alstonine and biological macromolecules were first investigated to provide the rationale for preferential labelling. Molecular filtration and spectrosfluorometric techniques with different macromolecules and isopolynucleotides have demonstrated that binding occurs only in the presence of uridyl rings. For the binding affect only the fluorescence intensity of alstonine it can be assumed that it involves only the side chain of the fluorescent compound. The capability for preferential staining was verified in culture using SK-OV-3 cells and rat hepatocarcinoma cells as tumor cells and Mouse fibroblasts or rat liver cells as controls. Techniques of image analysis have demonstrated the efficiency of cellular labelling even in aggregates of rat hepatocarcinoma. These experiments lead the way to the detection of tiny tumors developed on thin visceral walls, using a fiber optic device.

Community detection for fluorescent lifetime microscopy image segmentation

NASA Astrophysics Data System (ADS)

Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Achilefu, Samuel; Nussinov, Zohar

2014-03-01

Multiresolution community detection (CD) method has been suggested in a recent work as an efficient method for performing unsupervised segmentation of fluorescence lifetime (FLT) images of live cell images containing fluorescent molecular probes.1 In the current paper, we further explore this method in FLT images of ex vivo tissue slices. The image processing problem is framed as identifying clusters with respective average FLTs against a background or "solvent" in FLT imaging microscopy (FLIM) images derived using NIR fluorescent dyes. We have identified significant multiresolution structures using replica correlations in these images, where such correlations are manifested by information theoretic overlaps of the independent solutions ("replicas") attained using the multiresolution CD method from different starting points. In this paper, our method is found to be more efficient than a current state-of-the-art image segmentation method based on mixture of Gaussian distributions. It offers more than 1:25 times diversity based on Shannon index than the latter method, in selecting clusters with distinct average FLTs in NIR FLIM images.

Fluorescence correlation spectroscopy analysis for accurate determination of proportion of doubly labeled DNA in fluorescent DNA pool for quantitative biochemical assays.

PubMed

Hou, Sen; Sun, Lili; Wieczorek, Stefan A; Kalwarczyk, Tomasz; Kaminski, Tomasz S; Holyst, Robert

2014-01-15

Fluorescent double-stranded DNA (dsDNA) molecules labeled at both ends are commonly produced by annealing of complementary single-stranded DNA (ssDNA) molecules, labeled with fluorescent dyes at the same (3' or 5') end. Because the labeling efficiency of ssDNA is smaller than 100%, the resulting dsDNA have two, one or are without a dye. Existing methods are insufficient to measure the percentage of the doubly-labeled dsDNA component in the fluorescent DNA sample and it is even difficult to distinguish the doubly-labeled DNA component from the singly-labeled component. Accurate measurement of the percentage of such doubly labeled dsDNA component is a critical prerequisite for quantitative biochemical measurements, which has puzzled scientists for decades. We established a fluorescence correlation spectroscopy (FCS) system to measure the percentage of doubly labeled dsDNA (PDL) in the total fluorescent dsDNA pool. The method is based on comparative analysis of the given sample and a reference dsDNA sample prepared by adding certain amount of unlabeled ssDNA into the original ssDNA solution. From FCS autocorrelation functions, we obtain the number of fluorescent dsDNA molecules in the focal volume of the confocal microscope and PDL. We also calculate the labeling efficiency of ssDNA. The method requires minimal amount of material. The samples have the concentration of DNA in the nano-molar/L range and the volume of tens of microliters. We verify our method by using restriction enzyme Hind III to cleave the fluorescent dsDNA. The kinetics of the reaction depends strongly on PDL, a critical parameter for quantitative biochemical measurements. Copyright © 2013 Elsevier B.V. All rights reserved.

Structure-matched Phthalocyanine Ion Pair as a Red-emitting Fluorescent Optical Probe for the Analysis of Sodium Dodecylbenzenesulfonate with High Specificity and Sensitivity.

PubMed

Yu, Fei; Guo, Menglin; Deng, Yabin; Lu, Yin; Chen, Lin; Huang, Ping; Li, Donghui

2016-01-01

We have found that a positively charged cationic copper phthalocyanine, Alcian blue (Alcian blue 8GX), can efficiently quench the fluorescence of an oppositely charged red fluorescent phthalocyanine compound with a matched molecular structure, tetrasulfonated aluminum phthalocyanine (AlS4Pc), because of the formation of an ion pair complex (AlS4Pc-Alcian blue 8GX) that exhibits almost no fluorescence. An investigation was carried out on the fluorescence recovery of AlS4Pc-Alcian blue 8GX caused by a series of anionic surfactants containing a sulfonic group (sodium dodecylbenzenesulfonate (SDBS), sodium lauryl sulfate (SLS), and sodium dodecyl sulfate (SDS)). The results showed that SDBS exhibited a significant response, and the highest sensitivity among the surfactants. Due to its high efficiency of fluorescence quenching and the high level of fluorescence recovery, direct observes can even be performed by the naked eye. The results revealed that the Alcian blue 8GX-AlS4Pc ion-pair complex fluorescent probe only responded to SDBS in the low-concentration range. Based on the new founding, this study proposed a novel principle and method of fluorescence enhancement to specifically measure the concentration of SDBS, thereby achieving a highly sensitive and highly specific determination of SDBS. Under the optimal conditions, the fluorescence intensity (I(f)) of the system and the concentration of SDBS in the range of 1 × 10(-7) - 1 × 10(-5) mol/dm(3) exhibited a good linear relationship. This method is highly sensitive, and the operation is simple and rapid. It had been applied for the quantitative analysis of SDBS in environmental water, while achieving satisfactory results compared with those of the standard method. This study developed a new application of the fluorescent phthalocyanine compounds used as molecular probes in analytical sciences.

Multi-Channel Hyperspectral Fluorescence Detection Excited by Coupled Plasmon-Waveguide Resonance

PubMed Central

Du, Chan; Liu, Le; Zhang, Lin; Guo, Jun; Guo, Jihua; Ma, Hui; He, Yonghong

2013-01-01

We propose in this paper a biosensor scheme based on coupled plasmon-waveguide resonance (CPWR) excited fluorescence spectroscopy. A symmetrical structure that offers higher surface electric field strengths, longer surface propagation lengths and depths is developed to support guided waveguide modes for the efficient excitation of fluorescence. The optimal parameters for the sensor films are theoretically and experimentally investigated, leading to a detection limit of 0.1 nM (for a Cy5 solution). Multiplex analysis possible with the fluorescence detection is further advanced by employing the hyperspectral fluorescence technique to record the full spectra for every pixel on the sample plane. We demonstrate experimentally that highly overlapping fluorescence (Cy5 and Dylight680) can be distinguished and ratios of different emission sources can be determined accurately. This biosensor shows great potential for multiplex detections of fluorescence analytes. PMID:24129023

Highly Efficient Thermally Activated Delayed Fluorescence from an Excited-State Intramolecular Proton Transfer System

PubMed Central

2017-01-01

Thermally activated delayed fluorescence (TADF) materials have shown great potential for highly efficient organic light-emitting diodes (OLEDs). While the current molecular design of TADF materials primarily focuses on combining donor and acceptor units, we present a novel system based on the use of excited-state intramolecular proton transfer (ESIPT) to achieve efficient TADF without relying on the well-established donor–acceptor scheme. In an appropriately designed acridone-based compound with intramolecular hydrogen bonding, ESIPT leads to separation of the highest occupied and lowest unoccupied molecular orbitals, resulting in TADF emission with a photoluminescence quantum yield of nearly 60%. High external electroluminescence quantum efficiencies of up to 14% in OLEDs using this emitter prove that efficient triplet harvesting is possible with ESIPT-based TADF materials. This work will expand and accelerate the development of a wide variety of TADF materials for high performance OLEDs. PMID:28776019

Molecular approaches to third generation photovoltaics: photochemical up-conversion

NASA Astrophysics Data System (ADS)

Cheng, Yuen Yap; Fückel, Burkhard; Roberts, Derrick A.; Khoury, Tony; Clady, Rapha"l. G. C. R.; Tayebjee, Murad J. Y.; Piper, Roland; Ekins-Daukes, N. J.; Crossley, Maxwell J.; Schmidt, Timothy W.

2010-08-01

We have investigated a photochemical up-conversion system comprising a molecular mixture of a palladium porphyrin to harvest light, and a polycyclic aromatic hydrocarbon to emit light. The energy of harvested photons is stored as molecular triplet states which then annihilate to bring about up-converted fluorescence. The limiting efficiency of such triplet-triplet annihilation up-conversion has been believed to be 11% for some time. However, by rigorously investigating the kinetics of delayed fluorescence following pulsed excitation, we demonstrate instantaneous annihilation efficiencies exceeding 40%, and limiting efficiencies for the current system of ~60%. We attribute the high efficiencies obtained to the electronic structure of the emitting molecule, which exhibits an exceptionally high T2 molecular state. We utilize the kinetic data obtained to model an up-converting layer irradiated with broadband sunlight, finding that ~3% efficiencies can be obtained with the current system, with this improving dramatically upon optimization of various parameters.

Model of a thin film optical fiber fluorosensor

NASA Technical Reports Server (NTRS)

Egalon, Claudio O.; Rogowski, Robert S.

1991-01-01

The efficiency of core-light injection from sources in the cladding of an optical fiber is modeled analytically by means of the exact field solution of a step-profile fiber. The analysis is based on the techniques by Marcuse (1988) in which the sources are treated as infinitesimal electric currents with random phase and orientation that excite radiation fields and bound modes. Expressions are developed based on an infinite cladding approximation which yield the power efficiency for a fiber coated with fluorescent sources in the core/cladding interface. Marcuse's results are confirmed for the case of a weakly guiding cylindrical fiber with fluorescent sources uniformly distributed in the cladding, and the power efficiency is shown to be practically constant for variable wavelengths and core radii. The most efficient fibers have the thin film located at the core/cladding boundary, and fibers with larger differences in the indices of refraction are shown to be the most efficient.

A Micro Fluorescent Activated Cell Sorter for Astrobiology Applications

NASA Technical Reports Server (NTRS)

Platt, Donald W.; Hoover, Richard B.

2009-01-01

A micro-scale Fluorescent Activated Cell Sorter (microFACS) for astrobiology applications is under development. This device is designed to have a footprint of 7 cm x 7 cm x 4 cm and allow live-dead counts and sorting of cells that have fluorescent characteristics from staining. The FACS system takes advantage of microfluidics to create a cell sorter that can fit in the palm of the hand. A micron-scale channel allows cells to pass by a blue diode which causes emission of marker-expressed cells which are detected by a filtered photodetector. A small microcontroller then counts cells and operates high speed valves to select which chamber the cell is collected in (a collection chamber or a waste chamber). Cells with the expressed characteristic will be collected in the collection chamber. This system has been built and is currently being tested. We are also designing a system with integrated MEMS-based pumps and valves for a small and compact unit to fly on small satellite-based biology experiments.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hill, Steven C.; Williamson, Chatt C.; Doughty, David C.

This paper uses a mathematical model of fluorescent biological particles composed of bacteria and/or proteins (mostly as in Hill et al., 2013 [23]) to investigate the size-dependence of the total fluorescence emitted in all directions. The model applies to particles which have negligible reabsorption of fluorescence within the particle. The specific particles modeled here are composed of ovalbumin and of a generic Bacillus. The particles need not be spherical, and in some cases need not be homogeneous. However, the results calculated in this paper are for spherical homogeneous particles. Light absorbing and fluorescing molecules included in the model are aminomore » acids, nucleic acids, and several coenzymes. Here the excitation wavelength is 266 nm. The emission range, 300 to 370 nm, encompasses the fluorescence of tryptophan. The fluorescence cross section (C F) is calculated and compared with one set of published measured values. We investigate power law (Ad y) approximations to C F, where d is diameter, and A and y are parameters adjusted to fit the data, and examine how y varies with d and composition, including the fraction as water. The particle's fluorescence efficiency (Q F=C F/geometric-cross-section) can be written for homogeneous particles as Q absR F, where Q abs is the absorption efficiency, and R F, the fraction of the absorbed light emitted as fluorescence, is independent of size and shape. When Q F is plotted vs. m id or mi(m r-1)d, where m=m r+im i is the complex refractive index, the plots for different fractions of water in the particle tend to overlap.« less

CH3 NH3 PbBr3 Perovskite Nanocrystals as Efficient Light-Harvesting Antenna for Fluorescence Resonance Energy Transfer.

PubMed

Muthu, Chinnadurai; Vijayan, Anuja; Nair, Vijayakumar C

2017-05-04

Hybrid perovskites have created enormous research interest as a low-cost material for high-performance photovoltaic devices, light-emitting diodes, photodetectors, memory devices and sensors. Perovskite materials in nanocrystal form that display intense luminescence due to the quantum confinement effect were found to be particularly suitable for most of these applications. However, the potential use of perovskite nanocrystals as a light-harvesting antenna for possible applications in artificial photosynthesis systems is not yet explored. In the present work, we study the light-harvesting antenna properties of luminescent methylammonium lead bromide (CH 3 NH 3 PbBr 3 )-based perovskite nanocrystals using fluorescent dyes (rhodamine B, rhodamine 101, and nile red) as energy acceptors. Our studies revealed that CH 3 NH 3 PbBr 3 nanocrystals are an excellent light-harvesting antenna, and efficient fluorescence resonance energy transfer occurs from the nanocrystals to fluorescent dyes. Further, the energy transfer efficiency is found to be highly dependent on the number of anchoring groups and binding ability of the dyes to the surface of the nanocrystals. These observations may have significant implications for perovskite-based light-harvesting devices and their possible use in artificial photosynthesis systems. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Modulation of ICT probability in bi(polyarene)-based O-BODIPYs: towards the development of low-cost bright arene-BODIPY dyads.

PubMed

Gartzia-Rivero, Leire; Sánchez-Carnerero, Esther M; Jiménez, Josue; Bañuelos, Jorge; Moreno, Florencio; Maroto, Beatriz L; López-Arbeloa, Iñigo; de la Moya, Santiago

2017-09-12

We report the synthesis, and spectroscopic and electrochemical properties of a selected library of novel spiranic O-BODIPYs bearing a phenol-based bi(polyarene) unit tethered to the boron center through oxygen atoms. These dyes constitute an interesting family of arene-BODIPY dyads useful for the development of photonic applications due to their synthetic accessibility and tunable photonic properties. It is demonstrated that the electron-donor capability of the involved arene moiety switches on a non-emissive intramolecular charge transfer (ICT) state, which restricts the fluorescence efficiency of the dyad. Interestingly, the influence of this non-radiative deactivation channel can be efficiently modulated by the substitution pattern, either at the dipyrrin ligand or at the polyarene moiety. Thus, dyads featuring electron-rich dipyrrin and electron-poor polyarene show lower or almost negligible ICT probability, and hence display bright fluorescence upon dual excitation at far-away spectral regions. This synthetic approach has allowed the easy development of low-cost efficient ultraviolet-absorbing visible-emitting cassettes by selecting properly the substitution pattern of the involved key units, dipyrrin and bi(polyarene), to modulate not only absorption and emission wavelengths, but also fluorescence efficiencies.

Optimization of hybrid blue organic light-emitting diodes based on singlet and triplet exciton diffusion length

NASA Astrophysics Data System (ADS)

Lee, Song Eun; Lee, Ho Won; Lee, Jae Woo; Hwang, Kyo Min; Park, Soo Na; Yoon, Seung Soo; Kim, Young Kwan

2015-06-01

The hybrid blue organic light-emitting diodes (HB OLEDs) with triplet harvesting (TH) structures within an emitting layer (EML) are fabricated with fluorescent and phosphorescent EMLs. The TH is to transfer triplet excitons from fluorescence to phosphorescence, where they can decay radiatively. Remarkably, the half-decay lifetime of a hybrid blue device with fluorescent and phosphorescent EML thickness of 5 and 25 nm, measured at an initial luminance of 500 cd/m2, has improved twice than that of using a conventional structure. Additionally, the blue device’s efficiency improved. We attribute this improvement to the efficient triplet excitons energy transfer and the optimized distribution of the EML which depends on singlet and triplet excitons diffusion length that occurs within each the EML.

Fluorescence Correlation Spectroscopy to find the critical balance between extracellular association and intracellular dissociation of mRNA-complexes.

PubMed

Zhang, Heyang; De Smedt, Stefaan C; Remaut, Katrien

2018-05-10

Fluorescence Correlation Spectroscopy (FCS) is a promising tool to study interactions on a single molecule level. The diffusion of fluorescent molecules in and out of the excitation volume of a confocal microscope leads to the fluorescence fluctuations that give information on the average number of fluorescent molecules present in the excitation volume and their diffusion coefficients. In this context, we complexed mRNA into lipoplexes and polyplexes and explored the association/dissociation degree of complexes by using gel electrophoresis and FCS. FCS enabled us to measure the association and dissociation degree of mRNA-based complexes both in buffer and protein-rich biological fluids such as human serum and ascitic fluid, which is a clear advantage over gel electrophoresis that was only applicable in protein-free buffer solutions. Furthermore, following the complex stability in buffer and biological fluids by FCS assisted to understand how complex characteristics, such as charge ratio and strength of mRNA binding, correlated to the transfection efficiency. We found that linear polyethyleneimine prevented efficient translation of mRNA, most likely due to a too strong mRNA binding, whereas the lipid based carrier Lipofectamine ® messengerMAX did succeed in efficient release and subsequent translation of mRNA in the cytoplasm of the cells. Overall, FCS is a reliable tool for the in depth characterization of mRNA complexes and can help us to find the critical balance keeping mRNA bound in complexes in the extracellular environment and efficient intracellular mRNA release leading to protein production. The delivery of messenger RNA (mRNA) to cells is promising to treat a variety of diseases. Therefore, the mRNA is typically packed in small lipid particles or polymer particles that help the mRNA to reach the cytoplasm of the cells. These particles should bind and carry the mRNA in the extracellular environment (e.g. blood, peritoneal fluid, ...), but should release the mRNA again in the intracellular environment. In this paper, we evaluated a method (Fluorescence Correlation Spectroscopy) that allows for the in depth characterization of mRNA complexes and can help us to find the critical balance keeping mRNA bound in complexes in the extracellular environment and efficient intracellular mRNA release leading to protein production. Copyright © 2018. Published by Elsevier Ltd.

Creation of High Efficient Firefly Luciferase

NASA Astrophysics Data System (ADS)

Nakatsu, Toru

Firefly emits visible yellow-green light. The bioluminescence reaction is carried out by the enzyme luciferase. The bioluminescence of luciferase is widely used as an excellent tool for monitoring gene expression, the measurement of the amount of ATP and in vivo imaging. Recently a study of the cancer metastasis is carried out by in vivo luminescence imaging system, because luminescence imaging is less toxic and more useful for long-term assay than fluorescence imaging by GFP. However the luminescence is much dimmer than fluorescence. Then bioluminescence imaging in living organisms demands the high efficient luciferase which emits near infrared lights or enhances the emission intensity. Here I introduce an idea for creating the high efficient luciferase based on the crystal structure.

Forster resonance energy transfer in the system of human serum albumin-xanthene dyes

NASA Astrophysics Data System (ADS)

Kochubey, V. I.; Pravdin, A. B.; Melnikov, A. G.; Konstantinova, I.; Alonova, I. V.

2016-04-01

The processes of interaction of fluorescent probes: eosin and erythrosine with human serum albumin (HSA) were studied by the methods of absorption and fluorescence spectroscopy. Extinction coefficients of probes were determined. Critical transfer radius and the energy transfer efficiency were defined by fluorescence quenching of HSA. Analysis of the excitation spectra of HSA revealed that the energy transfer process is carried out mainly between tryptophanyl and probes.

Comment on ’Single Pentacene Molecules Detected by Fluorescence Excitation in a P-Terphenyl Crystal’

DTIC Science & Technology

1990-12-10

8217 NO 11 TITLE (include Security Classification) Comment on "Single Pentacene Molecules Detected by Fluorescence Excitation in a p-Terphenyl Crystal" 12...8217 {Continue on reverse it necessary and identify by block numboer) Using h--,Ihly efficient Fluorescence excitation spectroscov of individual pentacene ...molecular impurities in p-terphenvl crystals, we have observed that some pentacene defects exhibit spcntaneous spectral jumps in their resonance frequency at

Baculovirus-mediated gene transfer in butterfly wings in vivo: an efficient expression system with an anti-gp64 antibody

PubMed Central

2013-01-01

Background Candidate genes for color pattern formation in butterfly wings have been known based on gene expression patterns since the 1990s, but their functions remain elusive due to a lack of a functional assay. Several methods of transferring and expressing a foreign gene in butterfly wings have been reported, but they have suffered from low success rates or low expression levels. Here, we developed a simple, practical method to efficiently deliver and express a foreign gene using baculovirus-mediated gene transfer in butterfly wings in vivo. Results A recombinant baculovirus containing a gene for green fluorescent protein (GFP) was injected into pupae of the blue pansy butterfly Junonia orithya (Nymphalidae). GFP fluorescence was detected in the pupal wings and other body parts of the injected individuals three to five days post-injection at various degrees of fluorescence. We obtained a high GFP expression rate at relatively high virus titers, but it was associated with pupal death before color pattern formation in wings. To reduce the high mortality rate caused by the baculovirus treatment, we administered an anti-gp64 antibody, which was raised against baculovirus coat protein gp64, to infected pupae after the baculovirus injection. This treatment greatly reduced the mortality rate of the infected pupae. GFP fluorescence was observed in pupal and adult wings and other body parts of the antibody-treated individuals at various degrees of fluorescence. Importantly, we obtained completely developed wings with a normal color pattern, in which fluorescent signals originated directly from scales or the basal membrane after the removal of scales. GFP fluorescence in wing tissues spatially coincided with anti-GFP antibody staining, confirming that the fluorescent signals originated from the expressed GFP molecules. Conclusions Our baculovirus-mediated gene transfer system with an anti-gp64 antibody is reasonably efficient, and it can be an invaluable tool to transfer, express, and functionally examine foreign genes in butterfly wings and also in other non-model insect systems. PMID:23522444

Polydopamine nanotube mediated fluorescent biosensor for Hg(ii) determination through exonuclease III-assisted signal amplification.

PubMed

A, Ravikumar; P, Panneerselvam

2018-05-29

We describe a highly sensitive fluorescence biosensor incorporating polydopamine nanotubes (PDNTs) based on the mechanism of exonuclease III (Exo III) assisted signal amplification for the determination of Hg2+ in aqueous solution. Fluorescent probes of FAM labeled ssDNA (FAM-ssDNA) adsorbed on the PDNTs act as an efficient quencher. In the presence of Hg2+, the FAM-ssDNA can bind to Hg2+ to form double stranded DNA (dsDNA) via the formation of T-Hg2+-T base pairs. Then, the dsDNA was removed from the surface of the PDNTs to restore the fluorescence. The release of the dsDNA was triggered by Exo III digestion. At the same time, the liberated Hg2+ mediates a new cycle of digestion. This assay is ultrasensitive for the selective recognition of Hg2+, and a detection limit as low as 10 pM was achieved. In addition, the fluorescent biosensing system also displays remarkable specificity to Hg2+ in the presence of other possible competing ions. This approach was applied to the determination of Hg2+ in real water samples with good recovery and high efficiency for practical analysis.

Efficient Simulation of Secondary Fluorescence Via NIST DTSA-II Monte Carlo.

PubMed

Ritchie, Nicholas W M

2017-06-01

Secondary fluorescence, the final term in the familiar matrix correction triumvirate Z·A·F, is the most challenging for Monte Carlo models to simulate. In fact, only two implementations of Monte Carlo models commonly used to simulate electron probe X-ray spectra can calculate secondary fluorescence-PENEPMA and NIST DTSA-II a (DTSA-II is discussed herein). These two models share many physical models but there are some important differences in the way each implements X-ray emission including secondary fluorescence. PENEPMA is based on PENELOPE, a general purpose software package for simulation of both relativistic and subrelativistic electron/positron interactions with matter. On the other hand, NIST DTSA-II was designed exclusively for simulation of X-ray spectra generated by subrelativistic electrons. NIST DTSA-II uses variance reduction techniques unsuited to general purpose code. These optimizations help NIST DTSA-II to be orders of magnitude more computationally efficient while retaining detector position sensitivity. Simulations execute in minutes rather than hours and can model differences that result from detector position. Both PENEPMA and NIST DTSA-II are capable of handling complex sample geometries and we will demonstrate that both are of similar accuracy when modeling experimental secondary fluorescence data from the literature.

Activatable thermo-sensitive ICG encapsulated pluronic nanocapsules for temperature sensitive fluorescence tomography

NASA Astrophysics Data System (ADS)

Kwong, Tiffany C.; Nouizi, Farouk; Sampathkumaran, Uma; Zhu, Yue; Alam, Maksudul M.; Gulsen, Gultekin

2015-03-01

Fluorescent tomography has been hindered by poor tissue penetration and weak signal which results in poor spatial resolution and quantification accuracy. Recently, it has been reported that activatable temperature responsive fluorescent probes which respond to focused ultrasound heating can improve the resolution and quantification of fluorescent tomography in deep tissue. This has lead to a new imaging modality, "Temperature-modulated fluorescent tomography." This technique relies on activatable thermo-sensitive fluorescent nanocapsules for whose fluorescence quantum efficiency is temperature dependent. Within a 4-5° C temperature range, the fluorescent signal increase more than 10-fold. In this molecular probe, Indocyanine Green (ICG) is encapsulated inside the core of a thermo-reversible pluronic micelle. Here we show the fluorescence response and temperature range of the nanocapsules which have been optimized for a higher temperature range to be used for in vivo animal imaging. We report on the feasibility of these temperature-sensitive reversible nanocapsules for in vivo applications by studying the pharmacokinetics in a subcutaneous mouse tumor model in vivo.

Plasmonic enhancement of cyanine dyes for near-infrared light-triggered photodynamic/photothermal therapy and fluorescent imaging

NASA Astrophysics Data System (ADS)

Lu, Mindan; Kang, Ning; Chen, Chuan; Yang, Liuqing; Li, Yang; Hong, Minghui; Luo, Xiangang; Ren, Lei; Wang, Xiumin

2017-11-01

Near-infrared (NIR) triggered cyanine dyes have attracted considerable attention in multimodal tumor theranostics. However, NIR cyanine dyes used in tumor treatment often suffer from low fluorescence intensity and weak singlet oxygen generation efficiency, resulting in inadequate diagnostic and therapy efficacy for tumors. It is still a great challenge to improve both the photodynamic therapy (PDT) and fluorescent imaging (FLI) efficacy of cyanine dyes in tumor applications. Herein, a novel multifunctional nanoagent AuNRs@SiO2-IR795 was developed to realize the integrated photothermal/photodynamic therapy (PTT/PDT) and FLI at a very low dosage of IR795 (0.4 μM) based on metal-enhanced fluorescence (MEF) effects. In our design, both the fluorescence intensity and reactive oxygen species of AuNRs@SiO2-IR795 nanocomposites were significantly enhanced up to 51.7 and 6.3 folds compared with free IR795, owing to the localized surface plasmon resonance band of AuNRs overlapping with the absorption or fluorescence emission band of the IR795 dye. Under NIR laser irradiation, the cancer cell inhibition efficiency in vitro with synergetic PDT/PTT was up to 82.3%, compared with 10.3% for free IR795. Moreover, the enhanced fluorescence intensity of our designed nanocomposites was helpful to track their behavior in tumor cells. Therefore, our designed nanoagents highlight the applications of multimodal diagnostics and therapy in tumors based on MEF.

Interaction and energy transfer studies between bovine serum albumin and CdTe quantum dots conjugates: CdTe QDs as energy acceptor probes.

PubMed

Kotresh, M G; Inamdar, L S; Shivkumar, M A; Adarsh, K S; Jagatap, B N; Mulimani, B G; Advirao, G M; Inamdar, S R

2017-06-01

In this paper, a systematic investigation of the interaction of bovine serum albumin (BSA) with water-soluble CdTe quantum dots (QDs) of two different sizes capped with carboxylic thiols is presented based on steady-state and time-resolved fluorescence measurements. Efficient Förster resonance energy transfer (FRET) was observed to occur from BSA donor to CdTe acceptor as noted from reduction in the fluorescence of BSA and enhanced fluorescence from CdTe QDs. FRET parameters such as Förster distance, spectral overlap integral, FRET rate constant and efficiency were determined. The quenching of BSA fluorescence in aqueous solution observed in the presence of CdTe QDs infers that fluorescence resonance energy transfer is primarily responsible for the quenching phenomenon. Bimolecular quenching constant (k q ) determined at different temperatures and the time-resolved fluorescence data provide additional evidence for this. The binding stoichiometry and various thermodynamic parameters are evaluated by using the van 't Hoff equation. The analysis of the results suggests that the interaction between BSA and CdTe QDs is entropy driven and hydrophobic forces play a key role in the interaction. Binding of QDs significantly shortened the fluorescence lifetime of BSA which is one of the hallmarks of FRET. The effect of size of the QDs on the FRET parameters are discussed in the light of FRET parameters obtained. Copyright © 2016 John Wiley & Sons, Ltd.

Generation and validation of homozygous fluorescent knock-in cells using CRISPR-Cas9 genome editing.

PubMed

Koch, Birgit; Nijmeijer, Bianca; Kueblbeck, Moritz; Cai, Yin; Walther, Nike; Ellenberg, Jan

2018-06-01

Gene tagging with fluorescent proteins is essential for investigations of the dynamic properties of cellular proteins. CRISPR-Cas9 technology is a powerful tool for inserting fluorescent markers into all alleles of the gene of interest (GOI) and allows functionality and physiological expression of the fusion protein. It is essential to evaluate such genome-edited cell lines carefully in order to preclude off-target effects caused by (i) incorrect insertion of the fluorescent protein, (ii) perturbation of the fusion protein by the fluorescent proteins or (iii) nonspecific genomic DNA damage by CRISPR-Cas9. In this protocol, we provide a step-by-step description of our systematic pipeline to generate and validate homozygous fluorescent knock-in cell lines.We have used the paired Cas9D10A nickase approach to efficiently insert tags into specific genomic loci via homology-directed repair (HDR) with minimal off-target effects. It is time-consuming and costly to perform whole-genome sequencing of each cell clone to check for spontaneous genetic variations occurring in mammalian cell lines. Therefore, we have developed an efficient validation pipeline of the generated cell lines consisting of junction PCR, Southern blotting analysis, Sanger sequencing, microscopy, western blotting analysis and live-cell imaging for cell-cycle dynamics. This protocol takes between 6 and 9 weeks. With this protocol, up to 70% of the targeted genes can be tagged homozygously with fluorescent proteins, thus resulting in physiological levels and phenotypically functional expression of the fusion proteins.

What to Do if a Compact Fluorescent Light (CFL) Bulb or Fluorescent Tube Light Bulb Breaks: Printable Instructions

EPA Pesticide Factsheets

The broken bulb can continue to release mercury vapor until it is cleaned up and removed. This cleanup guidance represents minimum recommended actions to reduce mercury exposure, and will be updated as more efficient practices are identified.

Sun-induced chlorophyll fluorescence, photosynthesis, and light use efficiency of a soybean field from seasonally continuous measurements

USDA-ARS?s Scientific Manuscript database

Recent development of sun-induced chlorophyll fluorescence (SIF) technology is stimulating studies to remotely approximate canopy photosynthesis (measured as gross primary production, GPP). While multiple applications have advanced the empirical relationship between GPP and SIF, mechanistic understa...

Absorption-emission optrode and methods of use thereof

DOEpatents

Hirschfeld, T.B.

1990-05-29

A method and apparatus are described for monitoring the physical and chemical properties of a sample fluid by measuring an optical signal generated by a fluorescent substance and modulated by an absorber substance. The emission band of the fluorescent substance overlaps the absorption band of the absorber substance, and the degree of overlap is dependent on the physical and chemical properties of the sample fluid. The fluorescent substance and absorber substance are immobilized on a substrate so that an effective number of molecules thereof are sufficiently close for resonant energy transfer to occur, thereby providing highly efficient modulation of the fluorescent emissions of the fluorescent substance by the absorber substance. 4 figs.

Visualization of nucleic acids with synthetic exciton-controlled fluorescent oligonucleotide probes.

PubMed

Wang, Dan Ohtan; Okamoto, Akimitsu

2015-01-01

Engineered probes to adapt new photochemical properties upon recognition of target nucleic acids offer powerful tools to DNA and RNA visualization technologies. Herein, we describe a rapid and effective visualization method of nucleic acids in both fixed and living cells with hybridization-sensitive fluorescent oligonucleotide probes. These probes are efficiently quenched in an aqueous environment due to the homodimeric, excitonic interactions between fluorophores but become highly fluorescent upon hybridization to DNA or RNA with complementary sequences. The fast hybridization kinetics and quick fluorescence activation of the new probes allow applications to simplify the conventional fluorescent in situ hybridization protocols and reduce the amount of time to process the samples. Furthermore, hybridization-sensitive fluorescence emission of the probes allows monitoring dynamic behaviors of RNA in living cells.

Enhancing early bladder cancer detection with fluorescence-guided endoscopic optical coherence tomography

NASA Astrophysics Data System (ADS)

Pan, Y. T.; Xie, T. Q.; Du, C. W.; Bastacky, S.; Meyers, S.; Zeidel, M. L.

2003-12-01

We report an experimental study of the possibility of enhancing early bladder cancer diagnosis with fluorescence-image-guided endoscopic optical coherence tomography (OCT). After the intravesical instillation of a 10% solution of 5-aminolevulinic acid, simultaneous fluorescence imaging (excitation of 380-420 nm, emission of 620-700 nm) and OCT are performed on rat bladders to identify the photochemical and morphological changes associated with uroepithelial tumorigenesis. The preliminary results of our ex vivo study reveal that both fluorescence and OCT can identify early uroepithelial cancers, and OCT can detect precancerous lesions (e.g., hyperplasia) that fluorescence may miss. This suggests that a cystoscope combining 5-aminolevulinic acid fluorescence and OCT imaging has the potential to enhance the efficiency and sensitivity of early bladder cancer diagnosis.

Interventional fluorescence spectroscopy: preliminary results to detect tumor margins during glioma resection with two fluorescence spectra of PpIX

NASA Astrophysics Data System (ADS)

Alston, L. M.; Guyotat, J.; Mahieu-Williame, L.; Hebert, M.; Kantapareddy, P.; Meyronet, D.; Rousseau, D.; Montcel, B.

2017-07-01

We show the feasibility of using an intraoperative spectroscopic device to identify tumors margins during glioma resection. The collected fluorescence spectra is fitted with two reference spectra of PpIX and the contribution of each spectrum enables to overcome the sensitivity of current techniques by seeing tumor margins and low grade gliomas.

Genetically encoded sensors and fluorescence microscopy for anticancer research

NASA Astrophysics Data System (ADS)

Zagaynova, Elena V.; Shirmanova, Marina V.; Sergeeva, Tatiana F.; Klementieva, Natalia V.; Mishin, Alexander S.; Gavrina, Alena I.; Zlobovskay, Olga A.; Furman, Olga E.; Dudenkova, Varvara V.; Perelman, Gregory S.; Lukina, Maria M.; Lukyanov, Konstantin A.

2017-02-01

Early response of cancer cells to chemical compounds and chemotherapeutic drugs were studied using novel fluorescence tools and microscopy techniques. We applied confocal microscopy, two-photon fluorescence lifetime imaging microscopy and super-resolution localization-based microscopy to assess structural and functional changes in cancer cells in vitro. The dynamics of energy metabolism, intracellular pH, caspase-3 activation during staurosporine-induced apoptosis as well as actin cytoskeleton rearrangements under chemotherapy were evaluated. We have showed that new genetically encoded sensors and advanced fluorescence microscopy methods provide an efficient way for multiparameter analysis of cell activities

X-ray Fluorescence Holography: Principles, Apparatus, and Applications

NASA Astrophysics Data System (ADS)

Hayashi, Kouichi; Korecki, Pawel

2018-06-01

X-ray fluorescence holography (XFH) is an atomic structure determination technique that combines the capabilities of X-ray diffraction and X-ray fluorescence spectroscopy. It provides a unique means of gaining fully three-dimensional information about the local atomic structure and lattice site positions of selected elements inside compound samples. In this work, we discuss experimental and theoretical aspects that are essential for the efficient recording and analysis of X-ray fluorescence holograms and review the most recent advances in XFH. We describe experiments performed with brilliant synchrotron radiation as well as with tabletop setups that employ conventional X-ray tubes.

Elemental analysis using a handheld X-Ray fluorescence spectrometer

USGS Publications Warehouse

Groover, Krishangi D.; Izbicki, John

2016-06-24

The U.S. Geological Survey is collecting geologic samples from local stream channels, aquifer materials, and rock outcrops for studies of trace elements in the Mojave Desert, southern California. These samples are collected because geologic materials can release a variety of elements to the environment when exposed to water. The samples are to be analyzed with a handheld X-ray fluorescence (XRF) spectrometer to determine the concentrations of up to 27 elements, including chromium.

Concentration, size, and excitation power effects on fluorescence from microdroplets and microparticles containing tryptophan and bacteria

NASA Astrophysics Data System (ADS)

Fell, Nicholas F., Jr.; Pinnick, Ronald G.; Hill, Steven C.; Videen, Gorden W.; Niles, Stanley; Chang, Richard K.; Holler, Stephen; Pan, Yongle; Bottiger, Jerold R.; Bronk, Burt V.

1999-01-01

Our group has been developing a system for single-particle fluorescence detection of aerosolized agents. This paper describes the most recent steps in the evolution of this system. The effects of fluorophore concentrations, droplet size, and excitation power have also been investigated with microdroplets containing tryptophan in water to determine the effects of these parameters on our previous results. The vibrating orifice droplet generator was chosen for this study base don its ability to generate particles of well- known and reproducible size. The power levels required to reach saturation and photodegradation were determined. In addition, the collection of fluorescence emission was optimized through the use of a UV achromatic photographic lens. This arrangement permitted collection of images of the droplet stream. Finally, the use of a dual-beam, conditional firing scheme facilitated the collection of improved signal- to-noise single-shot spectra from individual biological particles.

Ultrafast fluorescence spectroscopy of biologically relevant chromophores using type II difference frequency generation

NASA Astrophysics Data System (ADS)

Léonard, J.; Gelot, T.; Torgasin, K.; Haacke, S.

2011-01-01

A novel femtosecond fluorescence experiment based on type II difference frequency mixing is demonstrated. This approach is particularly interesting for near-UV emitting biological chromophores like amino acids and nucleotides, as the fluorescence is converted into the spectral range where CCD have their highest quantum efficiencies. The method is implemented with a 5-kHz amplified Ti:Sapphire laser system and first results obtained with 2,5-diphenyloxazole (PPO) in ethanol are reported.

Development of a dielectrophoresis-assisted surface plasmon resonance fluorescence biosensor for detection of bacteria

NASA Astrophysics Data System (ADS)

Kuroda, Chiaki; Iizuka, Ryota; Ohki, Yoshimichi; Fujimaki, Makoto

2018-05-01

To detect biological substances such as bacteria speedily and accurately, a dielectrophoresis-assisted surface plasmon resonance (SPR) fluorescence biosensor is being developed. Using Escherichia coli as a target organism, an appropriate voltage frequency to collect E. coli cells on indium tin oxide quadrupole electrodes by dielectrophoresis is analyzed. Then, E. coli is stained with 4‧,6-diamidino-2-phenylindole (DAPI). To clearly detect fluorescence signals from DAPI-stained E. coli cells, the sensor is optimized so that we can excite SPR on Al electrodes by illuminating 405 nm photons. As a result, the number of fluorescence signals is increased on the electrodes by the application of a low-frequency voltage. This indicates that E. coli cells with a lower permittivity than the surrounding water are collected by negative dielectrophoresis onto the electrodes where the electric field strength is lowest.

In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse

PubMed Central

McGinty, James; Stuckey, Daniel W.; Soloviev, Vadim Y.; Laine, Romain; Wylezinska-Arridge, Marzena; Wells, Dominic J.; Arridge, Simon R.; French, Paul M. W.; Hajnal, Joseph V.; Sardini, Alessandro

2011-01-01

Förster resonance energy transfer (FRET) is a powerful biological tool for reading out cell signaling processes. In vivo use of FRET is challenging because of the scattering properties of bulk tissue. By combining diffuse fluorescence tomography with fluorescence lifetime imaging (FLIM), implemented using wide-field time-gated detection of fluorescence excited by ultrashort laser pulses in a tomographic imaging system and applying inverse scattering algorithms, we can reconstruct the three dimensional spatial localization of fluorescence quantum efficiency and lifetime. We demonstrate in vivo spatial mapping of FRET between genetically expressed fluorescent proteins in live mice read out using FLIM. Following transfection by electroporation, mouse hind leg muscles were imaged in vivo and the emission of free donor (eGFP) in the presence of free acceptor (mCherry) could be clearly distinguished from the fluorescence of the donor when directly linked to the acceptor in a tandem (eGFP-mCherry) FRET construct. PMID:21750768

Synthesis and characterization of photoswitchable fluorescent silica nanoparticles.

PubMed

Fölling, Jonas; Polyakova, Svetlana; Belov, Vladimir; van Blaaderen, Alfons; Bossi, Mariano L; Hell, Stefan W

2008-01-01

We have designed and synthesized a new functional (amino reactive) highly efficient fluorescent molecular switch (FMS) with a photochromic diarylethene and a rhodamine fluorescent dye. The reactive group in this FMS -N-hydroxysuccinimide ester- allows selective labeling of amino containing molecules or other materials. In ethanolic solutions, the compound displays a large fluorescent quantum yield of 52 % and a large fluorescence modulation ratio (94 %) between two states that may be interconverted with red and near-UV light. Silica nanoparticles incorporating the new FMS were prepared and characterized, and their spectroscopic and switching properties were also studied. The dye retained its properties after the incorporation into the silica, thereby allowing light-induced reversible high modulation of the fluorescence signal of a single particle for up to 60 cycles, before undergoing irreversible photobleaching. Some applications of these particles in fluorescence microscopy are also demonstrated. In particular, subdiffraction images of nanoparticles were obtained, in the focal plane of a confocal microscope.

Laser-Induced Photofragmentation Fluorescence Imaging of Alkali Compounds in Flames.

PubMed

Leffler, Tomas; Brackmann, Christian; Aldén, Marcus; Li, Zhongshan

2017-06-01

Laser-induced photofragmentation fluorescence has been investigated for the imaging of alkali compounds in premixed laminar methane-air flames. An ArF excimer laser, providing pulses of wavelength 193 nm, was used to photodissociate KCl, KOH, and NaCl molecules in the post-flame region and fluorescence from the excited atomic alkali fragment was detected. Fluorescence emission spectra showed distinct lines of the alkali atoms allowing for efficient background filtering. Temperature data from Rayleigh scattering measurements together with simulations of potassium chemistry presented in literature allowed for conclusions on the relative contributions of potassium species KOH and KCl to the detected signal. Experimental approaches for separate measurements of these components are discussed. Signal power dependence and calculated fractions of dissociated molecules indicate the saturation of the photolysis process, independent on absorption cross-section, under the experimental conditions. Quantitative KCl concentrations up to 30 parts per million (ppm) were evaluated from the fluorescence data and showed good agreement with results from ultraviolet absorption measurements. Detection limits for KCl photofragmentation fluorescence imaging of 0.5 and 1.0 ppm were determined for averaged and single-shot data, respectively. Moreover, simultaneous imaging of KCl and NaCl was demonstrated using a stereoscope with filters. The results indicate that the photofragmentation method can be employed for detailed studies of alkali chemistry in laboratory flames for validation of chemical kinetic mechanisms crucial for efficient biomass fuel utilization.

Effect of lateral mobility of fluorescent probes in lipid mixing assays of cell fusion.

PubMed

Huang, S K; Cheng, M; Hui, S W

1990-11-01

Monolayers of human erythrocytes, immobilized on a cover slip, were induced to fuse by polyethylene glycol (mol wt 8,000). The mobility of fluorescent probes, 1-oleoyl-2-[12-[(7-nitro-2,1,3-benzoxadizol-4-yl)amino]dodecanoyl] phosphatidyl-choline (C12-NBD-PC), from labeled cells to unlabeled cells was monitored by video-enhanced fluorescence microscopy. A dequenching curve was obtained from the measurement of fluorescence intensities of pairs of fused cells over time. The dequenching curve and the curve obtained from macroscopic measurements of a cell monolayer (described in the preceding article) were compared and discussed. The slow probe transfer rate between a pair of fused cells was explained by a diffusion model based on membrane area conservation and the geometry of the fusion lumen. An equivalent lumen between two fused cells, thought to be the main rate limitation of probe mobility after fusion, was calculated to be approximately 130 nm in diameter. Lumens of 75 nm in diameter were observed by electron microscopy. Thus, the rate of macroscopic fluorescence dequenching depends not only upon the fusion efficiency, but also upon the number of simultaneous fusion partners, the geometry of their contact points, and the lateral mobility of the fluorescent probes through these points. The relative fusion efficiency can be derived only from the saturation dequenching values.

Acute toxicity of excess mercury on the photosynthetic performance of cyanobacterium, S. platensis--assessment by chlorophyll fluorescence analysis.

PubMed

Lu, C M; Chau, C W; Zhang, J H

2000-07-01

Measurement of chlorophyll fluorescence has been shown to be a rapid, non-invasive, and reliable method to assess photosynthetic performance in a changing environment. In this study, acute toxicity of excess Hg on the photosynthetic performance of the cyanobacterium S. platensis, was investigated by use of chlorophyll fluorescence analysis after cells were exposed to excess Hg (up to 20 microM) for 2 h. The results determined from the fast fluorescence kinetics showed that Hg induced a significant increase in the proportion of the Q(B)-non-reducing PSII reaction centers. The fluorescence parameters measured under the steady state of photosynthesis demonstrated that the increase of Hg concentration led to a decrease in the maximal efficiency of PSII photochemistry, the efficiency of excitation energy capture by the open PSII reaction centers, and the quantum yield of PSII electron transport. Mercury also resulted in a decrease in the coefficients of photochemical and non-photochemical quenching. Mercury may have an acute toxicity on cyanobacteria by inhibiting the quantum yield of photosynthesis sensitively and rapidly. Such changes occurred before any other visible damages that may be evaluated by other conventional measurements. Our results also demonstrated that chlorophyll fluorescence analysis can be used as a useful physiological tool to assess early stages of change in photosynthetic performance of algae in response to heavy metal pollution.

A simple and sensitive high-throughput GFP screening in woody and herbaceous plants.

PubMed

Hily, Jean-Michel; Liu, Zongrang

2009-03-01

Green fluorescent protein (GFP) has been used widely as a powerful bioluminescent reporter, but its visualization by existing methods in tissues or whole plants and its utilization for high-throughput screening remains challenging in many species. Here, we report a fluorescence image analyzer-based method for GFP detection and its utility for high-throughput screening of transformed plants. Of three detection methods tested, the Typhoon fluorescence scanner was able to detect GFP fluorescence in all Arabidopsis thaliana tissues and apple leaves, while regular fluorescence microscopy detected it only in Arabidopsis flowers and siliques but barely in the leaves of either Arabidopsis or apple. The hand-held UV illumination method failed in all tissues of both species. Additionally, the Typhoon imager was able to detect GFP fluorescence in both green and non-green tissues of Arabidopsis seedlings as well as in imbibed seeds, qualifying it as a high-throughput screening tool, which was further demonstrated by screening the seedlings of primary transformed T(0) seeds. Of the 30,000 germinating Arabidopsis seedlings screened, at least 69 GFP-positive lines were identified, accounting for an approximately 0.23% transformation efficiency. About 14,000 seedlings grown in 16 Petri plates could be screened within an hour, making the screening process significantly more efficient and robust than any other existing high-throughput screening method for transgenic plants.

Fluorescent biosensor for the detection of hyaluronidase: intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore.

PubMed

Chib, Rahul; Mummert, Mark; Bora, Ilkay; Laursen, Bo W; Shah, Sunil; Pendry, Robert; Gryczynski, Ignacy; Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal

2016-05-01

In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. Graphical Abstract Scheme showing cleavage of HA-ADOTA probe by hyaluronidase and the change in the emission spectrum of HA-ADOTA probe before and after cleavage by hyaluronidase.

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform

NASA Astrophysics Data System (ADS)

Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

2015-11-01

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr05839b

Improving confocal microscopy with solid-state semiconductor excitation sources

NASA Astrophysics Data System (ADS)

Sivers, Nelson L.

To efficiently excite the fluorescent dyes used in imaging biological samples with a confocal microscope, the wavelengths of the exciting laser must be near the fluorochrome absorption peak. However, this causes imaging problems when the fluorochrome absorption and emission spectra overlap significantly, i.e. have small Stokes shifts, which is the case for most fluorochromes that emit in the red to infrared. As a result, the reflected laser excitation cannot be distinguished from the information-containing fluorescence signal. However, cryogenically cooling the exciting laser diode enabled the laser emission wavelengths to be tuned to shorter wavelengths, decreasing the interference between the laser and the fluorochrome's fluorescence. This reduced the amount of reflected laser light in the confocal image. However, the cooled laser diode's shorter wavelength signal resulted in slightly less efficient fluorochrome excitation. Spectrophotometric analysis showed that as the laser diodes were cooled, their output power increased, which more than compensated for the lower fluorochrome excitation and resulted in significantly more intense fluorescence. Thus, by tuning the laser diode emission wavelengths away from the fluorescence signal, less reflected laser light and more fluorescence information reached the detector, creating images with better signal to noise ratios. Additionally, new, high, luminous flux, light-emitting diodes (LEDs) are now powerful enough to create confocal fluorescence signals comparable to those produced by the traditional laser excitation sources in fluorescence confocal microscopes. The broader LED spectral response effectively excited the fluorochrome, yet was spectrally limited enough for standard filter sets to separate the LED excitation from the fluorochrome fluorescence signal. Spectrophotometric analysis of the excitation and fluorescence spectra of several fluorochromes showed that high-powered, LED-induced fluorescence contained the same spectral information and could be more intense than that produced by lasers. An alternative, LED-based, confocal microscope is proposed in this thesis that would be capable of exciting multiple fluorochromes in a single specimen, producing images of several distinct cellular components simultaneously. The inexpensive, LED-based, confocal microscope would require lower peak excitation intensities to produce fluorescence signals equal to those produced by laser excitation, reducing cellular damage and slowing fluorochrome photobleaching.

Radioisotopic energy conversion system (RECS): A new radioisotopic power cell, based on nuclear, atomic, and radiation transport principles

NASA Astrophysics Data System (ADS)

Steinfelds, Eric Victor

The topic of this thesis is the development of the Radioisotope Energy Conversion System (RECS) in a project which is utilizing analytical computational assisted design and some experimental Research in the investigation of fluorescers and effective transducers with the appropriate energy range choice for the conversion of energy. It is desirable to increase the efficiency in electrical power from the raw kinetic power available from the radioactive material within radioisotope power generators. A major step in this direction is the development and use of Radioisotope Energy Conversion Systems to supplement and ideally replace Radioactive Thermal Generators (RTG). It is possible to achieve electrical conversion efficiencies exceeding 25% for RECS power devices compared to only 9 percent efficiency for RTG's. The theoretical basis with existent materials for the potential achievability of efficiencies above 25% is documented within this thesis. The fundamental RECS consists of a radioisotope radiative source (C1), a mediating fluorescent gas (C2) which readily absorbs energy from the beta particles (or alpha's) and subsequently emits blue or UV photons, photovoltaic cells (C3) to convert the blue and UV photons into electrical energy [2], and electrical circuitry (C4). Solid State inspired component (C3), due to its theoretical (and attainable) high efficiency, is a large step ahead of the RTG design concept. The radioisotope flux source produces the beta(-) particles or alpha particles. Geometrically, presently, we prefer to have the ambient fluorescent gas surround the radioisotope flux source. Our fluorescer shall be a gas such as Krypton. Our specifically wide band-gap photovoltaic cells shall have gap energies which are slightly less than that of UV photons produced by the fluorescing gas. Diamond and Aluminum Nitride sample materials are good potential choices for photovoltaic cells, as is explained here in. Out of the material examples discussed, the highest electric power to mass ratio is found to be readily attainable with strontium-90 as the radiative source. Krypton-85 is indisputably the most efficient in RECS devices. In the conclusion in chapter VI, suggestions are given on acceptable ways of containing krypton-85 and providing sufficient shielding on deep space probes destined to use krypton-85 powered 'batteries'.

Establishment of integrated protocols for automated high throughput kinetic chlorophyll fluorescence analyses.

PubMed

Tschiersch, Henning; Junker, Astrid; Meyer, Rhonda C; Altmann, Thomas

2017-01-01

Automated plant phenotyping has been established as a powerful new tool in studying plant growth, development and response to various types of biotic or abiotic stressors. Respective facilities mainly apply non-invasive imaging based methods, which enable the continuous quantification of the dynamics of plant growth and physiology during developmental progression. However, especially for plants of larger size, integrative, automated and high throughput measurements of complex physiological parameters such as photosystem II efficiency determined through kinetic chlorophyll fluorescence analysis remain a challenge. We present the technical installations and the establishment of experimental procedures that allow the integrated high throughput imaging of all commonly determined PSII parameters for small and large plants using kinetic chlorophyll fluorescence imaging systems (FluorCam, PSI) integrated into automated phenotyping facilities (Scanalyzer, LemnaTec). Besides determination of the maximum PSII efficiency, we focused on implementation of high throughput amenable protocols recording PSII operating efficiency (Φ PSII ). Using the presented setup, this parameter is shown to be reproducibly measured in differently sized plants despite the corresponding variation in distance between plants and light source that caused small differences in incident light intensity. Values of Φ PSII obtained with the automated chlorophyll fluorescence imaging setup correlated very well with conventionally determined data using a spot-measuring chlorophyll fluorometer. The established high throughput operating protocols enable the screening of up to 1080 small and 184 large plants per hour, respectively. The application of the implemented high throughput protocols is demonstrated in screening experiments performed with large Arabidopsis and maize populations assessing natural variation in PSII efficiency. The incorporation of imaging systems suitable for kinetic chlorophyll fluorescence analysis leads to a substantial extension of the feature spectrum to be assessed in the presented high throughput automated plant phenotyping platforms, thus enabling the simultaneous assessment of plant architectural and biomass-related traits and their relations to physiological features such as PSII operating efficiency. The implemented high throughput protocols are applicable to a broad spectrum of model and crop plants of different sizes (up to 1.80 m height) and architectures. The deeper understanding of the relation of plant architecture, biomass formation and photosynthetic efficiency has a great potential with respect to crop and yield improvement strategies.

Investigations regarding the wet decontamination of fluorescent lamp waste using iodine in potassium iodide solutions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tunsu, Cristian, E-mail: tunsu@chalmers.se; Ekberg, Christian; Foreman, Mark

Highlights: • A wet-based decontamination process for fluorescent lamp waste is proposed. • Mercury can be leached using iodine in potassium iodide solution. • The efficiency of the process increases with an increase in leachant concentration. • Selective leaching of mercury from rare earth elements is achieved. • Mercury is furthered recovered using ion exchange, reduction or solvent extraction. - Abstract: With the rising popularity of fluorescent lighting, simple and efficient methods for the decontamination of discarded lamps are needed. Due to their mercury content end-of-life fluorescent lamps are classified as hazardous waste, requiring special treatment for disposal. A simplemore » wet-based decontamination process is required, especially for streams where thermal desorption, a commonly used but energy demanding method, cannot be applied. In this study the potential of a wet-based process using iodine in potassium iodide solution was studied for the recovery of mercury from fluorescent lamp waste. The influence of the leaching agent’s concentration and solid/liquid ratio on the decontamination efficiency was investigated. The leaching behaviour of mercury was studied over time, as well as its recovery from the obtained leachates by means of anion exchange, reduction, and solvent extraction. Dissolution of more than 90% of the contained mercury was achieved using 0.025/0.05 M I{sub 2}/KI solution at 21 °C for two hours. The efficiency of the process increased with an increase in leachant concentration. 97.3 ± 0.6% of the mercury contained was dissolved at 21 °C, in two hours, using a 0.25/0.5 M I{sub 2}/KI solution and a solid to liquid ratio of 10% w/v. Iodine and mercury can be efficiently removed from the leachates using Dowex 1X8 anion exchange resin or reducing agents such as sodium hydrosulphite, allowing the disposal of the obtained solution as non-hazardous industrial wastewater. The extractant CyMe{sub 4}BTBP showed good removal of mercury, with an extraction efficiency of 97.5 ± 0.7% being achieved in a single stage. Better removal of mercury was achieved in a single stage using the extractants Cyanex 302 and Cyanex 923 in kerosene, respectively.« less

Carbon nanotube-mediated siRNA delivery for gene silencing in cancer cells

NASA Astrophysics Data System (ADS)

Hong, Tu; Guo, Honglian; Xu, Yaqiong

2011-10-01

Small interfering RNA (siRNA) is potentially a promising tool in influencing gene expression with a high degree of target specificity. However, its poor intracellular uptake, instability in vivo, and non-specific immune stimulations impeded its effect in clinical applications. In this study, carbon nanotubes (CNTs) functionalized with two types of phospholipid-polyethylene glycol (PEG) have shown capabilities to stabilize siRNA in cell culture medium during the transfection and efficiently deliver siRNA into neuroblastoma and breast cancer cells. Moreover, the intrinsic optical properties of CNTs have been investigated through absorption and fluorescence measurements. We have found that the directly-functionalized groups play an important role on the fluorescence imaging of functionalized CNTs. The unique fluorescence imaging and high delivery efficiency make CNTs a promising material to deliver drugs and evaluate the treatment effect simultaneously.

Multi-channel lock-in amplifier assisted femtosecond time-resolved fluorescence non-collinear optical parametric amplification spectroscopy with efficient rejection of superfluorescence background.

PubMed

Mao, Pengcheng; Wang, Zhuan; Dang, Wei; Weng, Yuxiang

2015-12-01

Superfluorescence appears as an intense background in femtosecond time-resolved fluorescence noncollinear optical parametric amplification spectroscopy, which severely interferes the reliable acquisition of the time-resolved fluorescence spectra especially for an optically dilute sample. Superfluorescence originates from the optical amplification of the vacuum quantum noise, which would be inevitably concomitant with the amplified fluorescence photons during the optical parametric amplification process. Here, we report the development of a femtosecond time-resolved fluorescence non-collinear optical parametric amplification spectrometer assisted with a 32-channel lock-in amplifier for efficient rejection of the superfluorescence background. With this spectrometer, the superfluorescence background signal can be significantly reduced to 1/300-1/100 when the seeding fluorescence is modulated. An integrated 32-bundle optical fiber is used as a linear array light receiver connected to 32 photodiodes in one-to-one mode, and the photodiodes are further coupled to a home-built 32-channel synchronous digital lock-in amplifier. As an implementation, time-resolved fluorescence spectra for rhodamine 6G dye in ethanol solution at an optically dilute concentration of 10(-5)M excited at 510 nm with an excitation intensity of 70 nJ/pulse have been successfully recorded, and the detection limit at a pump intensity of 60 μJ/pulse was determined as about 13 photons/pulse. Concentration dependent redshift starting at 30 ps after the excitation in time-resolved fluorescence spectra of this dye has also been observed, which can be attributed to the formation of the excimer at a higher concentration, while the blueshift in the earlier time within 10 ps is attributed to the solvation process.

Variability in chlorophyll fluorescence spectra of eggplant fruit grown under different light environments: a case study.

PubMed

Ospina Calvo, Brian; Parapugna, Tamara L; Lagorio, M Gabriela

2017-05-17

The main goal of the present work was to clarify physiological strategies in plants whose chloroplasts were developed under different light environments. The specific objective was to elucidate the influence of the spectral distribution of light on the chlorophyll fluorescence ratio and on photosynthetic parameters. To achieve this purpose, three species of eggplant fruit (black, purple and white striped and white) were used as a case study and their chlorophyll fluorescence was analyzed in detail. Spectra of the non-variable fluorescence in each part of the fruit were corrected for distortions by light reabsorption processes using a physical model. The main conclusion of this work was that the corrected fluorescence ratio was dependent on the contribution of each photosystem to the fluorescence and consequently on the environmental lighting conditions, becoming higher when illumination was rich in long wavelengths. Variable chlorophyll fluorescence, similar to that observed from plant leaves, was detected for the pulp of the black eggplant, for the pulp of the purple and white striped eggplant and for the intact fruit of the black eggplant. The maximum quantum efficiency of photosystem II in the light-adapted state (F' v /F' m ), the quantum efficiency of photosystem II (Φ PSII ), and the photochemical and non-photochemical quenching coefficients (qP and qNP/NPQ respectively) were determined in each case. The results could be explained very interestingly, in relation with the proportion of exciting light reaching each photosystem (I and II). The photochemical parameters obtained from variable chlorophyll fluorescence, allowed us to monitor non-destructively the physiological state of the black fruit during storage under both chilled or room-temperature conditions.

Multi-channel lock-in amplifier assisted femtosecond time-resolved fluorescence non-collinear optical parametric amplification spectroscopy with efficient rejection of superfluorescence background

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mao, Pengcheng; Wang, Zhuan; Dang, Wei

Superfluorescence appears as an intense background in femtosecond time-resolved fluorescence noncollinear optical parametric amplification spectroscopy, which severely interferes the reliable acquisition of the time-resolved fluorescence spectra especially for an optically dilute sample. Superfluorescence originates from the optical amplification of the vacuum quantum noise, which would be inevitably concomitant with the amplified fluorescence photons during the optical parametric amplification process. Here, we report the development of a femtosecond time-resolved fluorescence non-collinear optical parametric amplification spectrometer assisted with a 32-channel lock-in amplifier for efficient rejection of the superfluorescence background. With this spectrometer, the superfluorescence background signal can be significantly reduced to 1/300–1/100more » when the seeding fluorescence is modulated. An integrated 32-bundle optical fiber is used as a linear array light receiver connected to 32 photodiodes in one-to-one mode, and the photodiodes are further coupled to a home-built 32-channel synchronous digital lock-in amplifier. As an implementation, time-resolved fluorescence spectra for rhodamine 6G dye in ethanol solution at an optically dilute concentration of 10{sup −5}M excited at 510 nm with an excitation intensity of 70 nJ/pulse have been successfully recorded, and the detection limit at a pump intensity of 60 μJ/pulse was determined as about 13 photons/pulse. Concentration dependent redshift starting at 30 ps after the excitation in time-resolved fluorescence spectra of this dye has also been observed, which can be attributed to the formation of the excimer at a higher concentration, while the blueshift in the earlier time within 10 ps is attributed to the solvation process.« less

HaloTag technology for specific and covalent labeling of fusion proteins.

PubMed

Benink, Hélène A; Urh, Marjeta

2015-01-01

Appending proteins of interest to fluorescent protein tags such as GFP has revolutionized how proteins are studied in the cellular environment. Over the last few decades many varieties of fluorescent proteins have been generated, each bringing new capability to research. However, taking full advantage of standard fluorescent proteins with advanced and differential features requires significant effort on the part of the researcher. This approach necessitates that many genetic fusions be generated and confirmed to function properly in cells with the same protein of interest. To lessen this burden, a newer category of protein fusion tags termed "self-labeling protein tags" has been developed. This approach utilizes a single protein tag, the function of which can be altered by attaching various chemical moieties (fluorescent labels, affinity handles, etc.). In this way a single genetically encoded protein fusion can easily be given functional diversity and adaptability as supplied by synthetic chemistry. Here we present protein labeling methods using HaloTag technology; comprised of HaloTag protein and the collection of small molecules designed to bind it specifically and provide it with varied functionalities. For imaging purposes these small molecules, termed HaloTag ligands, contain distinct fluorophores. Due to covalent and rapid binding between HaloTag protein and its ligands, labeling is permanent and efficient. Many of these ligands have been optimized for permeability across cellular membranes allowing for live cell labeling and imaging analysis. Nonpermeable ligands have also been developed for specific labeling of surface proteins. Overall, HaloTag is a versatile technology that empowers the end user to label a protein of interest with the choice of different fluorophores while alleviating the need for generation of multiple genetic fusions.

Toward two-photon excited fluorescence lifetime endomicroscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Hage, Charles-Henri; Leclerc, Pierre; Fabert, Marc; Brevier, Julien; Habert, Rémi; Braud, Flavie; Kudlinski, Alexandre; Louradour, Frédéric

2017-02-01

Fluorescence lifetime imaging microscopy (FLIM) represents a powerful tool for biological studies. Endoscopic FLIM applied to the intracellular native biomarker NADH and FAD represents a promising mean for in vivo in situ malignant tissue diagnosis in the medical field. Else, 2-photon-excited fluorescence (2PEF) provides increased 3D resolution and imaging depth. But very few demonstrations about 2PEF lifetime measurement through a fiber have been reported and none about endoscopic 2P-FLIM through a practical fiber length (< 3m). Our group has recently demonstrated the possibility to efficiently deliver through a very long optical fiber the short and intense excitation pulses required for 2P-FLIM. Our goal is now to check that collecting fluorescence through the same endoscopic fiber does not deteriorate the lifetime measurement. Relying on the basis previously published in case of 1PEF by P. French and co-workers (J. Biophotonics, 2015), we have experimentally quantitatively evaluated the influence on the lifetime measurement of the fiber chromatic and intermodal dispersions. The main result is that the fiber contribution to the system impulse response function, even in the case of a 3-meter long double-clad optical fiber, does not hinder the separation between free and bound NADH states using FLIM. Related calibrations and measurements will be detailed. Ongoing experiments about the development of a 2P-FLIM endomicroscope on the basis of an previously reported 2P-endomicroscope (Ducourthial et al., Sc. Reports, 2015), used under various configurations (i.e. point measurement in the center of the 2P-endomicroscope image, averaged lifetime, binned endoscopic 2P-FLIM image), will be also presented.

Study of Sugar Cane Management Systems in Brazil Using Laser Induced Fluorescence

NASA Astrophysics Data System (ADS)

Cabral, Jader; Villas-Boas, Paulino; Carvalho, Camila; Corá, José Eduardo; Milori, Débora

2014-05-01

Brazil is the largest producer of cane sugar, consequently, is a leader in the production of bio-ethanol, a clean and renewable energy that fits the model of sustainable economy as discussed and pursued by our society. Our state of São Paulo concentrates 60% of national production, representing a sizeable share in the range of world production. All this economic potential is closely monitored by the scientific community, which develops numerous studies seeking an improvement in production efficiency and reduced environmental impacts caused by the planting. However, the study of soil samples, in plantation areas, demands results about the content and structural forms of organic matter (OM). Also, the soil carbon stocks depend on the type of management. Our goal is to study OM of soil samples from four sugar cane management systems: (i) unburned cane harvest, (ii) preharvest burned, (iii) addition of sugarcane bagasse ash and (iv) addition of residue from the extraction of sucrose, using Laser Induced Fluorescence Spectroscopy of solid state. All the emission spectra were acquired using the system called LIFS-405, which consists of a diode laser Coherent, model cube with excitation at 405 nm, maximum output power of 50mJ and a mini-spectrometer, Ocean Optics USB2000-high sensitivity, with range of 194-894 nm and a fiber-optic bundle design (six excitation fibers in a circular path and one central fiber the collect the fluorescence). In this work, we will present the preliminary results evolving the humification index (HLIFS) of soil OM and total carbon amount (TC) for the different types of management. HLIFS shows a close correlation with the humification index of humic acid in solution obtained by means 2D conventional fluorescence spectroscopy.

Improved Diffuse Fluorescence Flow Cytometer Prototype for High Sensitivity Detection of Rare Circulating Cells In Vivo

NASA Astrophysics Data System (ADS)

Pestana, Noah Benjamin

Accurate quantification of circulating cell populations is important in many areas of pre-clinical and clinical biomedical research, for example, in the study of cancer metastasis or the immune response following tissue and organ transplants. Normally this is done "ex-vivo" by drawing and purifying a small volume of blood and then analyzing it with flow cytometry, hemocytometry or microfludic devices, but the sensitivity of these techniques are poor and the process of handling samples has been shown to affect cell viability and behavior. More recently "in vivo flow cytometry" (IVFC) techniques have been developed where fluorescently-labeled cells flowing in a small blood vessel in the ear or retina are analyzed, but the sensitivity is generally poor due to the small sampling volume. To address this, our group recently developed a method known as "Diffuse Fluorescence Flow Cytometry" (DFFC) that allows detection and counting of rare circulating cells with diffuse photons, offering extremely high single cell counting sensitivity. In this thesis, an improved DFFC prototype was designed and validated. The chief improvements were three-fold, i) improved optical collection efficiency, ii) improved detection electronics, and iii) development of a method to mitigate motion artifacts during in vivo measurements. In combination, these improvements yielded an overall instrument detection sensitivity better than 1 cell/mL in vivo, which is the most sensitive IVFC system reported to date. Second, development and validation of a low-cost microfluidic device reader for analysis of ocular fluids is described. We demonstrate that this device has equivalent or better sensitivity and accuracy compared a fluorescence microscope, but at an order-of-magnitude reduced cost with simplified operation. Future improvements to both instruments are also discussed.

A Fusion Protein of the p53 Transaction Domain and the p53-Binding Domain of the Oncoprotein MdmX as an Efficient System for High-Throughput Screening of MdmX Inhibitors.

PubMed

Chen, Rong; Zhou, Jingjing; Qin, Lingyun; Chen, Yao; Huang, Yongqi; Liu, Huili; Su, Zhengding

2017-06-27

In nearly half of cancers, the anticancer activity of p53 protein is often impaired by the overexpressed oncoprotein Mdm2 and its homologue, MdmX, demanding efficient therapeutics to disrupt the aberrant p53-MdmX/Mdm2 interactions to restore the p53 activity. While many potent Mdm2-specific inhibitors have already undergone clinical investigations, searching for MdmX-specific inhibitors has become very attractive, requiring a more efficient screening strategy for evaluating potential scaffolds or leads. In this work, considering that the intrinsic fluorescence residue Trp23 in the p53 transaction domain (p53p) plays an important role in determining the p53-MdmX/Mdm2 interactions, we constructed a fusion protein to utilize this intrinsic fluorescence signal to monitor high-throughput screening of a compound library. The fusion protein was composed of the p53p followed by the N-terminal domain of MdmX (N-MdmX) through a flexible amino acid linker, while the whole fusion protein contained a sole intrinsic fluorescence probe. The fusion protein was then evaluated using fluorescence spectroscopy against model compounds. Our results revealed that the variation of the fluorescence signal was highly correlated with the concentration of the ligand within 65 μM. The fusion protein was further evaluated with respect to its feasibility for use in high-throughput screening using a model compound library, including controls. We found that the imidazo-indole scaffold was a bona fide scaffold for template-based design of MdmX inhibitors. Thus, the p53p-N-MdmX fusion protein we designed provides a convenient and efficient tool for high-throughput screening of new MdmX inhibitors. The strategy described in this work should be applicable for other protein targets to accelerate drug discovery.

Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence

NASA Astrophysics Data System (ADS)

Du Le, Vinh Nguyen; Patterson, Michael S.; Farrell, Thomas J.; Hayward, Joseph E.; Fang, Qiyin

2015-12-01

The ability to recover the intrinsic fluorescence of biological fluorophores is crucial to accurately identify the fluorophores and quantify their concentrations in the media. Although some studies have successfully retrieved the fluorescence spectral shape of known fluorophores, the techniques usually came with heavy computation costs and did not apply for strongly absorptive media, and the intrinsic fluorescence intensity and fluorophore concentration were not recovered. In this communication, an experimental approach was presented to recover intrinsic fluorescence and concentration of fluorescein in the presence of hemoglobin (Hb). The results indicated that the method was efficient in recovering the intrinsic fluorescence peak and fluorophore concentration with an error of 3% and 10%, respectively. The results also suggested that chromophores with irregular absorption spectra (e.g., Hb) have more profound effects on fluorescence spectral shape than chromophores with monotonic absorption and scattering spectra (e.g., black India ink and polystyrene microspheres).

Structured illumination microscopy for dual-modality 3D sub-diffraction resolution fluorescence and refractive-index reconstruction

PubMed Central

Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.

2017-01-01

Though structured illumination (SI) microscopy is a popular imaging technique conventionally associated with fluorescent super-resolution, recent works have suggested its applicability towards sub-diffraction resolution coherent imaging with quantitative endogenous biological contrast. Here, we demonstrate that SI can efficiently integrate together the principles of fluorescent super-resolution and coherent synthetic aperture to achieve 3D dual-modality sub-diffraction resolution, fluorescence and refractive-index (RI) visualizations of biological samples. We experimentally demonstrate this framework by introducing a SI microscope capable of 3D sub-diffraction resolution fluorescence and RI imaging, and verify its biological visualization capabilities by experimentally reconstructing 3D RI/fluorescence visualizations of fluorescent calibration microspheres as well as alveolar basal epithelial adenocarcinoma (A549) and human colorectal adenocarcinmoa (HT-29) cells, fluorescently stained for F-actin. This demonstration may suggest SI as an especially promising imaging technique to enable future biological studies that explore synergistically operating biophysical/biochemical and molecular mechanisms at sub-diffraction resolutions. PMID:29296504

An individually coated near-infrared fluorescent protein as a safe and robust nanoprobe for in vivo imaging

NASA Astrophysics Data System (ADS)

Yang, Yu; Xiang, Kun; Yang, Yi-Xin; Wang, Yan-Wen; Zhang, Xin; Cui, Yangdong; Wang, Haifang; Zhu, Qing-Qing; Fan, Liqiang; Liu, Yuanfang; Cao, Aoneng

2013-10-01

A prerequisite for in vivo fluorescence imaging is the safety of fluorescent probes. Among all fluorescent probes, fluorescent proteins (FPs) might be the safest ones, which have been widely used in biological sciences at the gene level. But FPs have not been used in vivo in the purified form yet due to the instability of proteins. Here, we individually coat near-infrared (NIR) FPs (NIRFPs) with a silica nanoshell, resulting in NIRFP@silica, one of the safest and brightest NIR fluorescent nanoprobes with a quantum yield of 0.33 for in vivo imaging. The silica shell not only protects NIRFPs from denaturation and metabolic digestion, but also enhances the quantum yield and photostability of the coated NIRFPs. When injected via the tail vein, NIRFP@silica NPs can distribute all over the mouse body, and then can be efficiently eliminated through urine in 24 h, demonstrating its potential applications as a safe and robust NIR fluorescence probe for whole body imaging.A prerequisite for in vivo fluorescence imaging is the safety of fluorescent probes. Among all fluorescent probes, fluorescent proteins (FPs) might be the safest ones, which have been widely used in biological sciences at the gene level. But FPs have not been used in vivo in the purified form yet due to the instability of proteins. Here, we individually coat near-infrared (NIR) FPs (NIRFPs) with a silica nanoshell, resulting in NIRFP@silica, one of the safest and brightest NIR fluorescent nanoprobes with a quantum yield of 0.33 for in vivo imaging. The silica shell not only protects NIRFPs from denaturation and metabolic digestion, but also enhances the quantum yield and photostability of the coated NIRFPs. When injected via the tail vein, NIRFP@silica NPs can distribute all over the mouse body, and then can be efficiently eliminated through urine in 24 h, demonstrating its potential applications as a safe and robust NIR fluorescence probe for whole body imaging. Electronic supplementary information (ESI) available: A chromatogram of APTS-NIRFP, a TEM image of 40 nm NIRFP@silica, dispersion stability of NIRFP@silica, more whole body fluorescent images, serum biochemical parameters, and optical images of HE stained organ slices. See DOI: 10.1039/c3nr02508j

WE-FG-BRA-04: A Portable Confocal Microscope to Image Live Cell Damage Response Induced by Therapeutic Radiation

DOE Office of Scientific and Technical Information (OSTI.GOV)

McFadden, C; Flint, D; Grosshans, D

Purpose: To construct a custom and portable fluorescence confocal laser-scanning microscope (FCLSM) that can be placed in the path of therapeutic radiation beams to study real-time radiation-induced damage response in live cells. Methods: We designed and constructed a portable FCLSM with three laser diodes for excitation (405, 488, and 635 nm). An objective lens focuses the excitation light and collects fluorescence from the sample. A pair of galvanometer mirrors scans/collects the laser beam/fluorescence along the focal plane (x/y-directions). A stepper motor stage scans in the axial direction and positions the x/y of the image field. Barrier filters and dichroic mirrorsmore » are used to route the spectral emission bands to the appropriate photodetector. An avalanche photodiode collects near-infrared fluorescence; a photodiode collects back-reflected 635 nm light; and a photomultiplier tube collects green fluorescence in the range of eGFP/eYFP. A 200-µm diameter pinhole was used to implement the confocal geometry for near-infrared and red channels and a 150-µm diameter pinhole for the green channel. Data acquisition and system control were achieved using a high-throughput data acquisition card. In-house software developed in LabVIEW was used to control the hardware, collect data from the photodetectors and reconstruct the confocal images. Results: 6 frames/s can be acquired for a 25 µm{sup 2} (128×128 pixels) field of view, visualizing the entire volume of the cell nucleus (∼10 µm depth) in <10 s. To demonstrate the usefulness of our FCLSM, we imaged gold nanoshells in live cells, radiation-induced damage in fibrosarcoma cells expressing eGFP tagged to a DNA repair protein, and neurons expressing eGFP. The system can also image particle tracks in fluorescent nuclear track detectors. Conclusion: We developed a versatile and portable FCLSM that allows radiobiology studies in live cells exposed to therapeutic radiation. The FCLSM can be placed in any vertical beam line for top-to-bottom exposures. This research was supported by the Sister Institution Network Fund and the Center for Radiation Oncology Research at The University of Texas MD Anderson Cancer Center and Cancer Prevention and Research Institute of Texas. Gabriel Sawakuchi has research support from Elekta Inc.« less

An efficient fluorescent single-particle position tracking system for long-term pulsed measurements of nitrogen-vacancy centers in diamond

NASA Astrophysics Data System (ADS)

Kim, Kiho; Yun, Jiwon; Lee, Donghyuck; Kim, Dohun

2018-02-01

A simple and convenient design enables real-time three-dimensional position tracking of nitrogen-vacancy (NV) centers in diamond. The system consists entirely of commercially available components (a single-photon counter, a high-speed digital-to-analog converter, a phase-sensitive detector-based feedback device, and a piezo stage), eliminating the need for custom programming or rigorous optimization processes. With a large input range of counters and trackers combined with high sensitivity of single-photon counting, high-speed position tracking (upper bound recovery time of 0.9 s upon 250 nm of step-like positional shift) not only of bright ensembles, but also of low-photon-collection-efficiency single to few NV centers (down to 103 s-1) is possible. The tracking requires position modulation of only 10 nm, which allows simultaneous position tracking and pulsed measurements in the long term. Therefore, this tracking system enables measuring a single-spin magnetic resonance and Rabi oscillations at a very high resolution even without photon collection optimization. The system is widely applicable to various fields related to NV center quantum manipulation research such as NV optical trapping, NV tracking in fluid dynamics, and biological sensing using NV centers inside a biological cell.

Capillary electrophoresis, a method for the determination of nucleic acid ligands covalently attached to quantum dots representing a donor of Förster resonance energy transfer.

PubMed

Datinská, Vladimíra; Klepárník, Karel; Belšánová, Barbora; Minárik, Marek; Foret, František

2018-05-09

The synthesis and determination of the structure of a Förster resonance energy transfer probe intended for the detection of specific nucleic acid sequences are described here. The probe is based on the hybridization of oligonucleotide modified quantum dots with a fluorescently labeled nucleic acid sample resulting in changes of the fluorescence emission due to the energy transfer effect. The stoichiometry distribution of oligonucleotides conjugated to quantum dots was determined by capillary electrophoresis separation. The results indicate that one to four molecules of oligonucleotide are conjugated to the surface of a single nanoparticle. This conclusion is confirmed by the course of the dependence of Förster resonance energy transfer efficiency on the concentration of fluorescently labeled complementary single-stranded nucleic acid, showing saturation. While the energy transfer efficiency of the probe hybridized with complementary nucleic acid strands was 30%, negligible efficiency was observed with a non-complementary strands. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Study on the blocking effect of a quantum-dot TiO2 compact layer in dye-sensitized solar cells with ionic liquid electrolyte under low-intensity illumination

NASA Astrophysics Data System (ADS)

Zhai, Peng; Lee, Hyeonseok; Huang, Yu-Ting; Wei, Tzu-Chien; Feng, Shien-Ping

2016-10-01

In this study, ultrasmall and ultrafine TiO2 quantum dots (QDs) were prepared and used as a high-performance compact layer (CL) in dye-sensitized solar cells (DSCs). We systematically investigated the performance of TiO2 CL under both low-intensity light and indoor fluorescent light illumination and found that the efficiency of DSCs with the insertion of optimal TiO2 QDs-CL was increased up to 18.3% under indoor T5 fluorescent light illumination (7000 lux). We clarified the controversy over the blocking effect of TiO2 CL for the efficiency increment and confirmed that the TiO2 QDs-CL performed significantly better under low-intensity illumination due to the efficient suppression of electron recombination at the FTO/electrolyte interface. We, for the first time, demonstrate this potential for the application of the DSCs with TiO2 QDs-CL in the low-intensity light and indoor fluorescent light illumination.

In vivo wide-field multispectral scanning laser ophthalmoscopy–optical coherence tomography mouse retinal imager: longitudinal imaging of ganglion cells, microglia, and Müller glia, and mapping of the mouse retinal and choroidal vasculature

PubMed Central

Zhang, Pengfei; Zam, Azhar; Jian, Yifan; Wang, Xinlei; Li, Yuanpei; Lam, Kit S.; Burns, Marie E.; Sarunic, Marinko V.; Pugh, Edward N.; Zawadzki, Robert J.

2015-01-01

Abstract. Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) provide complementary views of the retina, with the former collecting fluorescence data with good lateral but relatively low-axial resolution, and the latter collecting label-free backscattering data with comparable lateral but much higher axial resolution. To take maximal advantage of the information of both modalities in mouse retinal imaging, we have constructed a compact, four-channel, wide-field (∼50  deg) system that simultaneously acquires and automatically coregisters three channels of confocal SLO and Fourier domain OCT data. The scanner control system allows “zoomed” imaging of a region of interest identified in a wide-field image, providing efficient digital sampling and localization of cellular resolution features in longitudinal imaging of individual mice. The SLO is equipped with a “flip-in” spectrometer that enables spectral “fingerprinting” of fluorochromes. Segmentation of retina layers and en face display facilitate spatial comparison of OCT data with SLO fluorescence patterns. We demonstrate that the system can be used to image an individual retinal ganglion cell over many months, to simultaneously image microglia and Müller glia expressing different fluorochromes, to characterize the distinctive spatial distributions and clearance times of circulating fluorochromes with different molecular sizes, and to produce unequivocal images of the heretofore uncharacterized mouse choroidal vasculature. PMID:26677070

Canopy Level Solar Induced Fluorescence for Vegetation in Controlled Experiments

NASA Technical Reports Server (NTRS)

Middleton, E. M.; Corp, L. A.; Campbell, P. K. Entcheva

2007-01-01

Solar induced chlorophyll fluorescence (SIF) was retrieved from high resolution reflectance spectra acquired one meter above saplings of three deciduous tree species during springtime (three weeks after leaf flush) and in late summer when foliage was mature. SIF was determined by application of the Fraunhofer Line Depth (FLD) Principal to above-canopy spectra acquired with an Analytical Spectral Devices (ASD) Fieldspec spectroradiometer (3.2 nm resolution with 1.2 nm sampling interval). SIF retrievals were made at the two atmospheric oxygen (O2) absorption features that occur in the chlorophyll fluorescence (ChlF) region (660 -780 nm). These telluric features are 02V, the broader and deeper feature centered at 760 nm, but located on the shoulder of the far-red ChlF peak at 740 nm; and 023, a narrow feature centered at 688 nm that is positioned near the red ChlF peak at 685 nm. Supporting, coincident leaf level fluorescence, reflectance, photochemical and other measurements were also made. At the leaf level, these measurements included in situ photosynthetic capacity (Pmax) and light adapted total chlorophyll fluorescence (Fs') collected at steady state under high light and controlled chamber conditions (e.g., temperature, PAR, humidity, and COz); optical properties (reflectance, transmittance, absorptance); chlorophyll and carotenoid content; specific leaf mass; carbon (C) and nitrogen (N) content; fluorescence emission spectra at multiple excitation wavelengths; the ChlF contribution to red (R) and far-red (FR) reflectance; fluorescence imagery; and fluorescence excitation-emission matrices (EEMs). The tree species examined were tulip poplar (Liriodendron tulipifera L.), red maple (Acer rubrum L.), and sweetgum (Liquidambar styraczflua L.), and each had been provided four levels of N augmentation (0, 19, 37, and 75 kg Nhectare seasonally) to simulate atmospheric deposition from air pollution. Whole-plant SIF measurements of these species were compared with SIF estimates derived using FluorMOD, a radiative transfer model that includes fluorescence properties of foliage, when provided with our supporting measurements. Simulated values for SIF were also compared with similar estimates made over three years for corn (Zea mays L) crops under N treatments (20, 50,70, and 140 kg Nhectare) and with corn and pepper (Capsicum annuum L.) plants provided dimethyl urea (DCMU, 0 and 5 x 10" M) which were grown in pots and grouped in artificial canopies for spectral measurements. For near-surface measurements of healthy and stressed vegetation, red SIF retrieved at 688 nm (023) varied between 2-7 mW/m(exp 2)/nm/sr while far-red SIF retrieved at 760 nm (O2A) varied between 0.5 and 4.0 mW/m(exp 2)/nm/sr. Typical values for the SIF red/far-red ratio ranged between 1.75 and 4.0. Relationships of SIF to spectral reflectance indices and foliar photochemical indices were examined, such as photosynthetic light use efficiency. Initial investigations of the variability in these measurements at the same leaf temperature for sunlit and shaded foliage showed us that shaded foliage produced higher ChlF and light use efficiency. Other factors that affect SIF determinations are discussed. These results will assist in determine the expected intensity of the SIF signal from vegetation near the surface, as well as to identify enhancements needed for FluorMOD (or other such models), and will assist in determining relationships of SIF to reflectance indices and carbon dynamics.

One-Pot Synthesis of Fe3O4@PS@P(AEMH-FITC) Magnetic Fluorescent Nanocomposites for Bimodal Imaging.

PubMed

Wang, Xuandong; Liu, Huiyu; Jun, Ren; Fu, Changhui; Li, Linlin; Li, Tianlong; Tang, Fangqiong; Meng, Xianwei

2016-03-01

Magnetic fluorescent nanocomposites have attracted much attention because of their merging magnetic and fluorescent properties for biomedical application. However, the procedure of synthesis of magnetic fluorescent nanocomposites is always complicated. In addition, the properties of fluorescent component could be easily influenced by magnetic component, retaining both of the magnetic and fluorescent properties into one single nanoparticle considered to be a significant challenge. Herein, we report one-pot method to synthesize multifunctional magnetic fluorescent Fe3O4@PS@P(AEMH-FITC) nanocomposites for bimodal imaging. The asprepared Fe3O4@PS@P(AEMH-FITC) nanocomposites with well-define spherical core/shell structure were stable properties. Moreover, the Fe3O4@PS@P(AEMH-FITC) nanocomposites displayed efficient fluorescent and magnetic properties, respectively. Meanwhile, the magnetic resonance imaging (MRI) and HePG2 cancer cell fluorescent images experiment results suggested that Fe3O4@PS@P(AEMH-FITC) nanocomposites could be used as MRI contrast agents and Fluorescence Imaging (FLI) agents for bioimaging application. Our investigation paves a facile avenue for synthesized magnetic fluorescent nanostructures with well biocompatibility for potential bioimaging application in MRI and FLI.

Influence of ZnO nanorod on the luminescent and electrical properties of fluorescent dye-doped polymer nanocomposite

NASA Astrophysics Data System (ADS)

Zhang, T.; Xu, Z.; Qian, L.; Tao, D. L.; Teng, F.; Xu, X. R.

2006-11-01

The luminescent properties of fluorescent dye-doped polymer dispersed with ZnO nanorods were investigated. Embedding ZnO nanorods in blend film results in a blue-shifted emission of fluorescent dye. It is accounted for in terms of the difference in permittivity between inorganic oxide nano-material and dye-doped polymer. Moreover, polymer light-emitting diodes with the addition of ZnO nanorods showed the lower threshold voltage and the higher charge current and electroluminescence efficiency.

Fluorescent labels and their use in separations

DOEpatents

Mathies, Richard A.; Glazer, Alexander; Ju, Jingyue

1997-01-01

Compositions are provided comprising sets of fluorescent labels carrying pairs of donor and acceptor dye molecules, designed for efficient excitation of the donors at a single wavelength and emission from the acceptor in each of the pairs at different wavelengths. The different molecules having different donor-acceptor pairs can be modified to have substantially the same mobility under separation conditions, by varying the distance between the donor and acceptor in a given pair. Particularly, the fluorescent compositions find use as labels in sequencing nucleic acids.

Fiber-optic multiphoton flow cytometry in whole blood and in vivo

NASA Astrophysics Data System (ADS)

Chang, Yu-Chung; Ye, Jing Yong; Thomas, Thommey P.; Cao, Zhengyi; Kotlyar, Alina; Tkaczyk, Eric R.; Baker, James R.; Norris, Theodore B.

2010-07-01

Circulating tumor cells in the bloodstream are sensitive indicators for metastasis and disease prognosis. Circulating cells have usually been monitored via extraction from blood, and more recently in vivo using free-space optics; however, long-term intravital monitoring of rare circulating cells remains a major challenge. We demonstrate the application of a two-photon-fluorescence optical fiber probe for the detection of cells in whole blood and in vivo. A double-clad fiber was used to enhance the detection sensitivity. Two-channel detection was employed to enable simultaneous measurement of multiple fluorescent markers. Because the fiber probe circumvents scattering and absorption from whole blood, the detected signal strength from fluorescent cells was found to be similar in phosphate-buffered saline (PBS) and in whole blood. The detection efficiency of cells labeled with the membrane-binding dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindoldicarbocyanine, 4-chlorobenzenesulfonate (DiD) was demonstrated to be the same in PBS and in whole blood. A high detection efficiency of green fluorescent protein (GFP)-expressing cells in whole blood was also demonstrated. To characterize in vivo detection, DiD-labeled untransfected and GFP-transfected cells were injected into live mice, and the cell circulation dynamics was monitored in real time. The detection efficiency of GFP-expressing cells in vivo was consistent with that observed ex vivo in whole blood.

Orientation dependence in fluorescent energy transfer between Cy3 and Cy5 terminally attached to double-stranded nucleic acids

PubMed Central

Iqbal, Asif; Arslan, Sinan; Okumus, Burak; Wilson, Timothy J.; Giraud, Gerard; Norman, David G.; Ha, Taekjip; Lilley, David M. J.

2008-01-01

We have found that the efficiency of fluorescence resonance energy transfer between Cy3 and Cy5 terminally attached to the 5′ ends of a DNA duplex is significantly affected by the relative orientation of the two fluorophores. The cyanine fluorophores are predominantly stacked on the ends of the helix in the manner of an additional base pair, and thus their relative orientation depends on the length of the helix. Observed fluorescence resonance energy transfer (FRET) efficiency depends on the length of the helix, as well as its helical periodicity. By changing the helical geometry from B form double-stranded DNA to A form hybrid RNA/DNA, a marked phase shift occurs in the modulation of FRET efficiency with helix length. Both curves are well explained by the standard geometry of B and A form helices. The observed modulation for both polymers is less than that calculated for a fully rigid attachment of the fluorophores. However, a model involving lateral mobility of the fluorophores on the ends of the helix explains the observed experimental data. This has been further modified to take account of a minor fraction of unstacked fluorophore observed by fluorescent lifetime measurements. Our data unequivocally establish that Förster transfer obeys the orientation dependence as expected for a dipole–dipole interaction. PMID:18676615

Spectroscopic insights on selfassembly and excited state interactions between rhodamine and phthalocyanine molecules.

PubMed

Geng, Hao; Zhang, Xian-Fu

2015-03-15

The absorption and fluorescence spectra as well as fluorescence lifetimes of tetrasulfonated zinc phthalocyanine ZnPc(SO3Na)4 were measured in the absence and presence of four rhodamine dyes, Rhodamine B (RB), Ethyl rhodamine B (ERB), Rhodamine 6G (R6G), Rhodamine 110 (R110), and Pyronine B (PYB). The ground state complexes of phthalocyanine-(Rhodamine)2 were observed which exhibit new absorption bands. The binding constants are all very large (0.86×10(5)-0.22×10(8) M(-1)), suggesting rhodamine-phthalocyanine pairs are very good combinations for efficient selfassembly. Both the fluorescence intensity and the lifetime values of ZnPc(SO3Na)4 were decreased by the presence of rhodamines. The structural effect of rhodamines on selfassembly is significant. The ground state binding and dynamic quenching capability is PYB>R6G>ERB>RB>R110. The dynamic fluorescence quenching is due to the photoinduced electron transfer (PET). The PET rate constant is very large and in the order of 10(13) M(-1) s(-1), much greater than kf and kic (in the order of 10(8) M(-1) s(-1)), which means that the PET efficiency is almost 100%. Therefore the non-covalent Pc-rhodamine is a very good pair of donor/acceptor for potential efficient solar energy conversion. Copyright © 2014 Elsevier B.V. All rights reserved.

Mirrored pyramidal wells for simultaneous multiple vantage point microscopy

PubMed Central

Seale, K.T.; Reiserer, R.S.; Markov, D.A.; Ges, I.A.; Wright, C.; Janetopoulos, C.; Wikswo, J.P.

2013-01-01

Summary We report a novel method for obtaining simultaneous images from multiple vantage points of a microscopic specimen using size-matched microscopic mirrors created from anisotropically etched silicon. The resulting pyramidal wells enable bright-field and fluorescent side-view images, and when combined with z-sectioning, provide additional information for 3D reconstructions of the specimen. We have demonstrated the 3D localization and tracking over time of the centrosome of a live Dictyostelium discoideum. The simultaneous acquisition of images from multiple perspectives also provides a five-fold increase in the theoretical collection efficiency of emitted photons, a property which may be useful for low-light imaging modalities such as bioluminescence, or low abundance surface-marker labelling. PMID:19017196

Realizing Highly Efficient Solution-Processed Homojunction-Like Sky-Blue OLEDs by Using Thermally Activated Delayed Fluorescent Emitters Featuring an Aggregation-Induced Emission Property.

PubMed

Wu, Kailong; Wang, Zian; Zhan, Lisi; Zhong, Cheng; Gong, Shaolong; Xie, Guohua; Yang, Chuluo

2018-04-05

Two new blue emitters, i.e., bis-[2-(9,9-dimethyl-9,10-dihydroacridine)-phenyl]-sulfone ( o-ACSO2) and bis-[3-(9,9-dimethyl-9,10-dihydroacridine)-phenyl]-sulfone ( m-ACSO2), with reserved fine thermally activated delayed fluorescent (TADF) nature and simply tuned thermal and optoelectronic properties, were synthesized by isomer engineering. The meta-linking compound, i.e., m-ACSO2, obtains the highest photoluminescence quantum yield with a small singlet-triplet energy gap, a moderate delayed fluorescent lifetime, excellent solubility, and neat film homogeneity. Due to its unique aggregation-induced emission (AIE) character, neat film-based heterojunction-like organic light-emitting diodes (OLEDs) are achievable. By inserting an excitonic inert exciton-blocking layer, the PN heterojunction-like emission accompanied by intefacial exciplex was shifted to a homojunction-like channel mainly from the AIE emitter itself, providing a new tactic to generate efficient blue color from neat films. The solution-processed nondoped sky-blue OLED employing m-ACSO2 as emitter with homojunction-like emission achieved a maximum external quantum efficiency of 17.2%. The design strategies presented herein provide practical methods to construct efficient blue TADF dyes and realize high-performance blue TADF devices.

Fluorescence turn-on detection of alkaline phosphatase activity based on controlled release of PEI-capped Cu nanoclusters from MnO2 nanosheets.

PubMed

Zhang, Yunyi; Li, Yongxin; Zhang, Cuiyun; Zhang, Qingfeng; Huang, Xinan; Yang, Meiding; Shahzad, Sohail Anjum; Lo, Kenneth Kam-Wing; Yu, Cong; Jiang, Shichun

2017-08-01

A fluorescence turn-on assay for alkaline phosphatase (ALP) activity is developed through the controlled release of polyethyleneimine-capped copper nanoclusters (PEI-capped CuNCs) from the MnO 2 nanosheets. In an aqueous solution, the positively charged PEI-capped CuNCs could be adsorbed onto the surface of the negatively charged MnO 2 nanosheets. Such adsorption through favorable electrostatic interactions could efficiently quench the nanocluster fluorescence emission via resonance energy transfer from the PEI-capped CuNCs to the MnO 2 nanosheets. 2-Phospho-L-ascorbic acid (AAP) could be hydrolyzed to L-ascorbic acid (AA) in the presence of ALP. AA could reduce MnO 2 into Mn 2+ and trigger the disintegration of the MnO 2 nanosheets. As a result, the CuNCs were released and the quenched fluorescence was recovered efficiently. The detection strategy is simple, inexpensive, sensitive, selective, with low toxicity, and has better biocompatibility. The newly fabricated biosensor for ALP activity will potentially make it a robust candidate for numerous biological and biomedical applications.

Enhancing the color gamut of white displays using novel deep-blue organic fluorescent dyes to form color-changed thin films with improved efficiency

NASA Astrophysics Data System (ADS)

Liu, Wei-Ting; Huang, Wen-Yao

2012-10-01

This study used the novel fluorescence based deep-blue-emitting molecule BPVPDA in an organic fluorescent color thin film to exhibit deep blue color with CIE coordinates of (0.13, 0.16). The developed original organic RGB color thin film technology enables the optimization of the distinctive features of an organic light emitting diode (OLED) and thin-film-transistor (TFT) LCD display. The color filter structure maintains the same high resolution to obtain a higher level of brightness in comparison with conventional organic RGB color thin film. The image-processing engine is designed to achieve a sharp text image for a TFT LCD with organic color thin films. The organic color thin films structure uses an organic dye dopant in a limpid photoresist. With this technology, the following characteristics can be obtained: 1. high color reproduction of gamut ratio, and 2. improved luminous efficiency with organic color fluorescent thin film. This performance is among the best results ever reported for a color-filter used on TFT-LCD or OLED.

Enhancing the color gamut of white displays using novel deep-blue organic fluorescent dyes to form color-changed thin films with improved efficiency

NASA Astrophysics Data System (ADS)

Liu, Wei-ting; Huang, Wen-Yao

2012-06-01

This study used novel fluorescence based deep-blue-emitting molecules, namely BPVPDA, an organic fluorescence color thin film using BPVPDA exhibit deep blue fluorine with CIE coordinates of (0.13,0.16). The developed original Organic RGB color thin film technology enables the optimization of the distinctive features of an organic light emitting diode (OLED) and (TFT) LCD display. The color filter structure maintains the same high resolution to obtain a higher level of brightness, in comparison with conventional organic RGB color thin film. The image-processing engine is designed to achieve a sharp text image for a thin-film-transistor (TFT) LCD with organic color thin films. The organic color thin films structure uses organic dye dopent in limpid photo resist. With this technology , the following characteristics can be obtained: (1) high color reproduction of gamut ratio, and (2) improved luminous efficiency with organic color fluorescence thin film. This performance is among the best results ever reported for a color-filter used on TFT-LCD and OLED.

Guidelines for application of fluorescent lamps in high-performance avionic backlight systems

NASA Astrophysics Data System (ADS)

Syroid, Daniel D.

1997-07-01

Fluorescent lamps have proven to be well suited for use in high performance avionic backlight systems as demonstrated by numerous production applications for both commercial and military cockpit displays. Cockpit display applications include: Boeing 777, new 737s, F-15, F-16, F-18, F-22, C- 130, Navy P3, NASA Space Shuttle and many others. Fluorescent lamp based backlights provide high luminance, high lumen efficiency, precision chromaticity and long life for avionic active matrix liquid crystal display applications. Lamps have been produced in many sizes and shapes. Lamp diameters range from 2.6 mm to over 20 mm and lengths for the larger diameter lamps range to over one meter. Highly convoluted serpentine lamp configurations are common as are both hot and cold cathode electrode designs. This paper will review fluorescent lamp operating principles, discuss typical requirements for avionic grade lamps, compare avionic and laptop backlight designs and provide guidelines for the proper application of lamps and performance choices that must be made to attain optimum system performance considering high luminance output, system efficiency, dimming range and cost.

NIR-induced highly sensitive detection of latent finger-marks by NaYF4:Yb,Er upconversion nanoparticles in a dry powder state

PubMed Central

Wang, Meng; Li, Ming; Yang, Mingying; Zhang, Xiaomei; Yu, Aoyang; Zhu, Ye; Qiu, Penghe; Mao, Chuanbin

2016-01-01

The most commonly found fingermarks at crime scenes are latent and, thus, an efficient method for detecting latent fingermarks is very important. However, traditional developing techniques have drawbacks such as low detection sensitivity, high background interference, complicated operation, and high toxicity. To tackle this challenge, we employed fluorescent NaYF4:Yb,Er upconversion nanoparticles (UCNPs), which can fluoresce visible light when excited by 980 nm human-safe near-infrared light, to stain the latent fingermarks on various substrate surfaces. The UCNPs were successfully used as a novel fluorescent label for the detection of latent fingermarks with high sensitivity, low background, high efficiency, and low toxicity on various substrates including non-infiltrating materials (glass, marble, aluminum alloy sheets, stainless steel sheets, aluminum foils, and plastic cards), semi-infiltrating materials (floor leathers, ceramic tiles, wood floor, and painted wood), and infiltrating materials such as various types of papers. This work shows that UCNPs are a versatile fluorescent label for the facile detection of fingermarks on virtually any material, enabling their practical applications in forensic sciences. PMID:27818741

A Neuron-Based Screening Platform for Optimizing Genetically-Encoded Calcium Indicators

PubMed Central

Schreiter, Eric R.; Hasseman, Jeremy P.; Tsegaye, Getahun; Fosque, Benjamin F.; Behnam, Reza; Shields, Brenda C.; Ramirez, Melissa; Kimmel, Bruce E.; Kerr, Rex A.; Jayaraman, Vivek; Looger, Loren L.; Svoboda, Karel; Kim, Douglas S.

2013-01-01

Fluorescent protein-based sensors for detecting neuronal activity have been developed largely based on non-neuronal screening systems. However, the dynamics of neuronal state variables (e.g., voltage, calcium, etc.) are typically very rapid compared to those of non-excitable cells. We developed an electrical stimulation and fluorescence imaging platform based on dissociated rat primary neuronal cultures. We describe its use in testing genetically-encoded calcium indicators (GECIs). Efficient neuronal GECI expression was achieved using lentiviruses containing a neuronal-selective gene promoter. Action potentials (APs) and thus neuronal calcium levels were quantitatively controlled by electrical field stimulation, and fluorescence images were recorded. Images were segmented to extract fluorescence signals corresponding to individual GECI-expressing neurons, which improved sensitivity over full-field measurements. We demonstrate the superiority of screening GECIs in neurons compared with solution measurements. Neuronal screening was useful for efficient identification of variants with both improved response kinetics and high signal amplitudes. This platform can be used to screen many types of sensors with cellular resolution under realistic conditions where neuronal state variables are in relevant ranges with respect to timing and amplitude. PMID:24155972

Rapid detection of bacterial pathogens using flourescence spectroscopy and chemometrics

USDA-ARS?s Scientific Manuscript database

This work presents the development of a method for rapid bacterial identification based on the fluorescence spectroscopy combined with multivariate analysis. Fluorescence spectra of pure three different genera of bacteria (Escherichia coli, Salmonella, and Campylobacter) were collected from 200...

Metal-enhanced fluorescence exciplex emission.

PubMed

Zhang, Yongxia; Mali, Buddha L; Geddes, Chris D

2012-01-01

In this letter, we report the first observation of metal-enhanced exciplex fluorescence, observed from anthracene in the presence of diethylaniline. Anthracene in the presence of diethylaniline in close proximity to Silver Island Films (SIFs) shows enhanced monomer and exciplex emission as compared to a non-silvered control sample containing no silver nanoparticles. Our findings suggest two complementary methods for the enhancement: (i) surface plasmons can radiate coupled monomer and exciplex fluorescence efficiently, and (ii) enhanced absorption (enhanced electric near-field) further facilitates enhanced emission. Our exciplex studies help us to further understand the complex photophysics of the metal-enhanced fluorescence technology. Copyright © 2011 Elsevier B.V. All rights reserved.

Fluorescence from a single Symbiodinium cell

NASA Astrophysics Data System (ADS)

Guzman, Christine; Han, Xue; Shoguchi, Eiichi; Chormaic, Síle Nic

2018-07-01

The partnership between coral and its algal symbionts, Symbiodinium, is crucial to the global environment. Yet, the regulatory process within the photosynthetic machinery of Symbiodinium is still not clearly understood. Here, we studied the influence of light stress from focussed red and blue lasers on single Symbiodinium cells. Fluorescence signals were measured to show cell response. Increasing the incident laser power or the exposure time resulted in an increase followed by a decline in fluorescence intensity. The trend of fluorescence intensity changes was associated with mechanisms of light use efficiency, non-photochemical quenching, photoinhibition, and repair of the cell. Our study provides new approaches to studying the photobiology and physiology of Symbiodinium cells.

Use of acousto-optic tunable filter in fluorescence imaging endoscopy

NASA Astrophysics Data System (ADS)

Bouhifd, Mounir; Whelan, Maurice; Aprahamian, Marc

2003-10-01

A prototype instrument for fluorescence-based medical diagnostics in vivo is described. The system consists of a rigid endoscope comprising a UV laser-source for fluorescence excitation and a white light source for direct imaging. An acousto-optic tuneable filter (AOTF) is employed as a full-field tuneable bandpass filter. This allows fast continuous or random-access tuning with high filtering efficiency. A study of the diagnostic potential of fluorescence imaging for pancreatitis was conducted on a rat model. In particular, the aim was to detect autofluorescence of endogenous protoporphyrin IX (PpIX) that has been shown to accumulate in early-stage diseased tissue undergoing an inflammatory response.

Fluorescence quenching by TEMPO: a sub-30 A single-molecule ruler.

PubMed

Zhu, Peizhi; Clamme, Jean-Pierre; Deniz, Ashok A

2005-11-01

A series of DNA molecules labeled with 5-carboxytetramethylrhodamine (5-TAMRA) and the small nitroxide radical TEMPO were synthesized and tested to investigate whether the intramolecular quenching efficiency can be used to measure short intramolecular distances in small ensemble and single-molecule experiments. In combination with distance calculations using molecular mechanics modeling, the experimental results from steady-state ensemble fluorescence and fluorescence correlation spectroscopy measurements both show an exponential decrease in the quenching rate constant with the dye-quencher distance in the 10-30 A range. The results demonstrate that TEMPO-5-TAMRA fluorescence quenching is a promising method to measure short distance changes within single biomolecules.

Detecting Circular RNAs by RNA Fluorescence In Situ Hybridization.

PubMed

Zirkel, Anne; Papantonis, Argyris

2018-01-01

Fluorescence in situ hybridization (FISH) coupled to high-resolution microscopy is a powerful method for analyzing the subcellular localization of RNA. However, the detection of circular RNAs (circRNAs) using microscopy is challenging because the only feature of a circRNA that can be used for the probe design is its junction. Circular RNAs are expressed at varying levels, and for their efficient monitoring by FISH, background fluorescence levels need to be kept low. Here, we describe a FISH protocol coupled to high-precision localizations using a single fluorescently labeled probe spanning the circRNA junction; this allows circRNA detection in mammalian cells with high signal-to-noise ratios.

Fluorescent porous silicon biological probes with high quantum efficiency and stability.

PubMed

Tu, Chang-Ching; Chou, Ying-Nien; Hung, Hsiang-Chieh; Wu, Jingda; Jiang, Shaoyi; Lin, Lih Y

2014-12-01

We demonstrate porous silicon biological probes as a stable and non-toxic alternative to organic dyes or cadmium-containing quantum dots for imaging and sensing applications. The fluorescent silicon quantum dots which are embedded on the porous silicon surface are passivated with carboxyl-terminated ligands through stable Si-C covalent bonds. The porous silicon bio-probes have shown photoluminescence quantum yield around 50% under near-UV excitation, with high photochemical and thermal stability. The bio-probes can be efficiently conjugated with antibodies, which is confirmed by a standard enzyme-linked immunosorbent assay (ELISA) method.

Efficient blue conversion from a 1064 nm microchip laser in long photonic crystal fiber tapers for fluorescence microscopy.

PubMed

Kudlinski, A; Lelek, M; Barviau, B; Audry, L; Mussot, A

2010-08-02

Using a low-cost microchip laser and a long photonic crystal fiber taper, we report a supercontinuum source with a very efficient visible conversion, especially in the blue region (around 420 nm). About 30 % of the total average output power is located in the 350-600 nm band, which is of primary importance in a number of biophotonics applications such as flow cytometry or fluorescence imaging microscopy for instance. We successfully demonstrate the use of this visible-enhanced source for a three-color imaging of HeLa cells in wide-field microscopy.

Observations of Ocean Primary Productivity Using MODIS

NASA Technical Reports Server (NTRS)

Esaias, Wayne E.; Abbott, Mark R.; Koblinsky, Chester J. (Technical Monitor)

2001-01-01

Measuring the magnitude and variability of oceanic net primary productivity (NPP) represents a key advancement toward our understanding of the dynamics of marine ecosystems and the role of the ocean in the global carbon cycle. MODIS observations make two new contributions in addition to continuing the bio-optical time series begun with Orbview-2's SeaWiFS sensor. First, MODIS provides weekly estimates of global ocean net primary productivity on weekly and annual time periods, and annual empirical estimates of carbon export production. Second, MODIS provides additional insight into the spatial and temporal variations in photosynthetic efficiency through the direct measurements of solar-stimulated chlorophyll fluorescence. The two different weekly productivity indexes (first developed by Behrenfeld & Falkowski and by Yoder, Ryan and Howard) are used to derive daily productivity as a function of chlorophyll biomass, incident daily surface irradiance, temperature, euphotic depth, and mixed layer depth. Comparisons between these two estimates using both SeaWiFS and MODIS data show significant model differences in spatial distribution after allowance for the different integration depths. Both estimates are strongly dependence on the accuracy of the chlorophyll determination. In addition, an empirical approach is taken on annual scales to estimate global NPP and export production. Estimates of solar stimulated fluorescence efficiency from chlorophyll have been shown to be inversely related to photosynthetic efficiency by Abbott and co-workers. MODIS provides the first global estimates of oceanic chlorophyll fluorescence, providing an important proof of concept. MODIS observations are revealing spatial patterns of fluorescence efficiency which show expected variations with phytoplankton photo-physiological parameters as measured during in-situ surveys. This has opened the way for research into utilizing this information to improve our understanding of oceanic NPP variability. Deriving the ocean bio-optical properties places severe demands on instrument performance (especially band to band precision) and atmospheric correction. Improvements in MODIS instrument characterization and calibration over the first 16 mission months have greatly improved the accuracy of the chlorophyll input fields and FLH, and therefore the estimates of NPP and fluorescence efficiency. Annual estimates now show the oceanic NPP accounts for 40-50% of the global total NPP, with significant interannual variations related to large scale ocean processes. Spatial variations in ocean NPP, and exported production, have significant effects on exchange of CO2 between the ocean and atmosphere. Further work is underway to improve both the primary productivity model functions, and to refine our understanding of the relationships between fluorescence efficiency and NPP estimates. We expect that the MODIS instruments will prove extremely useful in assessing the time dependencies of oceanic carbon uptake and effects of iron enrichment, within the global carbon cycle.

A novel cyanide-selective colorimetric and fluorescent chemosensor: first molecular security keypad lock based on phosphotungstic acid and CN- inputs.

PubMed

Tavallali, Hossein; Deilamy-Rad, Gohar; Parhami, Abolfath; Hasanli, Nahid

2014-02-15

Rhodamine B (RhB) an available dye has been developed as novel and efficient colorimetric and fluorometric chemosensor for cyanide ions in an absolutely aqueous media. The UV-vis absorption and fluorescent emission titrations experiments have been employed to study the sensing process. RhB could act as an efficient "ON-OFF" fluorescent response for phosphotungstic acid (H3PW12O40 or PTA) based on an ion associate process. Also (RhB(+))3 · PTA(3-) could operate as an "OFF-ON" fluorescent sensor for cyanide anions based on a ligand substitution process. It has been identified as highly sensitive probe for CN(-) which responds at 0.3 and 0.04 μmol L(-1) concentration levels by absorption and fluorescent method respectively. Depending upon the sequence of addition of PTA and CN(-) ions into the solution, RhB could be as a molecular security keypad lock with PTA and CN(-) inputs. The ionic inputs to new fluorophore have been mimicked as a superimposed electronic molecular keypad lock. The results were compared successfully (>96%) with the data of a spectrophotometry approved method (EPA 9014-1) for cyanide ions. Copyright © 2013 Elsevier B.V. All rights reserved.

System and method for monitoring cellular activity

NASA Technical Reports Server (NTRS)

Bearman, Gregory H. (Inventor); Fraser, Scott E. (Inventor); Lansford, Russell D. (Inventor)

2002-01-01

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

System and method for monitoring cellular activity

NASA Technical Reports Server (NTRS)

Bearman, Gregory H. (Inventor); Fraser, Scott E. (Inventor); Lansford, Russell D. (Inventor)

2004-01-01

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Highly efficient exciplex organic light-emitting diodes using thermally activated delayed fluorescent emitters as donor and acceptor materials.

PubMed

Jeon, Sang Kyu; Yook, Kyoung Soo; Lee, Jun Yeob

2016-06-03

Highly efficient exciplex type organic light-emitting diodes were developed using thermally activated delayed fluorescent emitters as donors and acceptors of an exciplex. Blue emitting bis[4-(9,9-dimethyl-9,10-dihydroacridine)phenyl]sulfone (DMAC-DPS) was a donor and 9,9'-(5-(4,6-diphenyl-1,3,5-triazin-2-yl)-1,3-phenylene)bis(9H-carbazole) (DDCzTrz) and 9,9',9″-(5-(4,6-diphenyl-1,3,5-triazin-2-yl)benzene-1,2,3-triyl)tris(9H-carbazole) (TCzTrz) were acceptor materials. The exciplexes of DMAC-DPS:TCzTrz and DMAC-DPS:DDCzTrz resulted in high photoluminescence quantum yield and high quantum efficiency in the green exciplex organic light-emitting diodes. High quantum efficiencies of 13.4% and 15.3% were obtained in the DMAC-DPS:DDCzTrz and DMAC-DPS:TCzTrz exciplex devices.

Highly efficient exciplex organic light-emitting diodes using thermally activated delayed fluorescent emitters as donor and acceptor materials

NASA Astrophysics Data System (ADS)

Jeon, Sang Kyu; Yook, Kyoung Soo; Lee, Jun Yeob

2016-06-01

Highly efficient exciplex type organic light-emitting diodes were developed using thermally activated delayed fluorescent emitters as donors and acceptors of an exciplex. Blue emitting bis[4-(9,9-dimethyl-9,10-dihydroacridine)phenyl]sulfone (DMAC-DPS) was a donor and 9,9‧-(5-(4,6-diphenyl-1,3,5-triazin-2-yl)-1,3-phenylene)bis(9H-carbazole) (DDCzTrz) and 9,9‧,9″-(5-(4,6-diphenyl-1,3,5-triazin-2-yl)benzene-1,2,3-triyl)tris(9H-carbazole) (TCzTrz) were acceptor materials. The exciplexes of DMAC-DPS:TCzTrz and DMAC-DPS:DDCzTrz resulted in high photoluminescence quantum yield and high quantum efficiency in the green exciplex organic light-emitting diodes. High quantum efficiencies of 13.4% and 15.3% were obtained in the DMAC-DPS:DDCzTrz and DMAC-DPS:TCzTrz exciplex devices.

Instantaneous and efficient surface wave excitation of a low pressure gas or gases

DOEpatents

Levy, Donald J.; Berman, Samuel M.

1988-01-01

A system for instantaneously ionizing and continuously delivering energy in the form of surface waves to a low pressure gas or mixture of low pressure gases, comprising a source of rf energy, a discharge container, (such as a fluorescent lamp discharge tube), an rf shield, and a coupling device responsive to rf energy from the source to couple rf energy directly and efficiently to the gas or mixture of gases to ionize at least a portion of the gas or gases and to provide energy to the gas or gases in the form of surface waves. The majority of the rf power is transferred to the gas or gases near the inner surface of the discharge container to efficiently transfer rf energy as excitation energy for at least one of the gases. The most important use of the invention is to provide more efficient fluorescent and/or ultraviolet lamps.

Apparatus to collect, classify, concentrate, and characterize gas-borne particles

DOEpatents

Rader, Daniel J.; Torczynski, John R.; Wally, Karl; Brockmann, John E.

2003-12-16

An aerosol lab-on-a-chip (ALOC) integrates one or more of a variety of particle collection, classification, concentration (enrichment), an characterization processes onto a single substrate or layered stack of such substrates. By mounting a UV laser diode laser light source on the substrate, or substrates tack, so that it is located down-stream of the sample inlet port and at right angle the sample particle stream, the UV light source can illuminate individual particles in the stream to induce a fluorescence response in those particles having a fluorescent signature such as biological particles, some of said particles. An illuminated particle having a fluorescent signal above a threshold signal would trigger a sorter module that would separate that particle from the particle stream.

Pyrimidine-based twisted donor-acceptor delayed fluorescence molecules: a new universal platform for highly efficient blue electroluminescence.

PubMed

Park, In Seob; Komiyama, Hideaki; Yasuda, Takuma

2017-02-01

Deep-blue emitters that can harvest both singlet and triplet excited states to give high electron-to-photon conversion efficiencies are highly desired for applications in full-color displays and white lighting devices based on organic light-emitting diodes (OLEDs). Thermally activated delayed fluorescence (TADF) molecules based on highly twisted donor-acceptor (D-A) configurations are promising emitting dopants for the construction of efficient deep-blue OLEDs. In this study, a simple and versatile D-A system combining acridan-based donors and pyrimidine-based acceptors has been developed as a new platform for high-efficiency deep-blue TADF emitters. The designed pre-twisted acridan-pyrimidine D-A molecules exhibit small singlet-triplet energy splitting and high photoluminescence quantum yields, functioning as efficient deep-blue TADF emitters. The OLEDs utilizing these TADF emitters display bright blue electroluminescence with external quantum efficiencies of up to 20.4%, maximum current efficiencies of 41.7 cd A -1 , maximum power efficiencies of 37.2 lm W -1 , and color coordinates of (0.16, 0.23). The design strategy featuring such acridan-pyrimidine D-A motifs can offer great prospects for further developing high-performance deep-blue TADF emitters and TADF-OLEDs.

Rapid fluorescence detection of pathogenic bacteria using magnetic enrichment technique combined with magnetophoretic chromatography.

PubMed

Che, Yulan; Xu, Yi; Wang, Renjie; Chen, Li

2017-08-01

A rapid and sensitive analytical method was developed to detect pathogenic bacteria which combined magnetic enrichment, fluorescence labeling with polyethylene glycol (PEG) magnetophoretic chromatography. As pathogenic bacteria usually exist in complex matrixes at low concentration, an efficient enrichment is essential for diagnosis. In order to capture series types of pathogenic bacteria in samples, amino-modified magnetic nanoparticles (Fe 3 O 4 @SiO 2 -NH 2 ) were prepared for efficient enrichment by the electrostatic interaction with pathogenic bacteria. It was shown that the capture efficiency reached up to 95.4% for Escherichia coli (E. coli). Furthermore, quantitative analysis of the bacteria was achieved by using acridine orange (AO) as a fluorescence probe for the captured E. coli due to its ability of staining series types of bacteria and rapid labeling. In order to remove the free magnetic nanoparticles and redundant fluorescent reagent, the labeled suspension was poured into a PEG separation column and was separated by applying an external magnetic field. The presence of 100 cfu mL -1 E. coli could be detected for semi-quantitative analysis by observing the separation column with the naked eye, and the concentration could be further evaluated by fluorescence detection. All the above processes were finished within 80 min. It was demonstrated that a good linear relationship existed between the fluorescence intensity and the concentration of E. coli ranging from 10 2 to 10 6  cfu mL -1 , with a detection limit of 100 cfu mL -1 when E. coli acted as target bacteria. The recovery rate of E. coli was 93.6∼102.0% in tap water and cooked meat samples, and the RSD was lower than 7% (n = 6); the result coincided with the conventional plate count method. Graphical abstract ᅟ.

High-Throughput Synthesis and Characterization of Eu Doped Ba xSr2- xSiO4 Thin Film Phosphors.

PubMed

Frost, Sara; Guérin, Samuel; Hayden, Brian E; Soulié, Jean-Philippe; Vian, Chris

2018-06-20

High-throughput techniques have been employed for the synthesis and characterization of thin film phosphors of Eu-doped Ba x Sr 2- x SiO 4 . Direct synthesis from evaporation of the constituent elements under a flux of atomic oxygen on a sapphire substrate at 850 °C was used to directly produce thin film libraries (415 nm thickness) of the crystalline orthosilicate phase with the desired compositional variation (0.24 > x > 1.86). The orthosilicate phase could be synthesized as a pure, or predominantly pure, phase. Annealing the as synthesized library in a reducing atmosphere resulted in the reduction of the Eu while retaining the orthosilicate phase, and resulted in a materials thin film library where fluorescence excited by blue light (450 nm) was observable by the naked eye. Parallel screening of the fluorescence from the combinatorial libraries of Eu doped Ba x Sr 2- x SiO 4 has been implemented by imaging the fluorescent radiation over the library using a monochrome digital camera using a series of color filters. Informatics tools have been developed to allow the 1931 CIE color coordinates and the relative quantum efficiencies of the materials library to be rapidly assessed and mapped against composition, crystal structure and phase purity. The range of compositions gave values of CIE x between 0.17 and 0.52 and CIE y between 0.48 and 0.69 with relative efficiencies in the range 2.0 × 10 -4 -7.6 × 10 -4 . Good agreement was obtained between the thin film phosphors and the fluorescence characteristics of a number of corresponding bulk phosphor powders. The thermal quenching of fluorescence in the thin film libraries was also measured in the temperature range 25-130 °C: The phase purity of the thin film was found to significantly influence both the relative quantum efficiency and the thermal quenching of the fluorescence.

Raman efficiencies of natural rocks and minerals: performance of a remote Raman system for planetary exploration at a distance of 10 meters.

PubMed

Stopar, Julie D; Lucey, Paul G; Sharma, Shiv K; Misra, Anupam K; Taylor, G Jeffrey; Hubble, Hugh W

2005-08-01

Raman spectroscopy is a powerful technique for materials analysis, and we are developing and analyzing a remote Raman system for use on a planetary lander or rover. We have acquired data at a distance of 10m from a variety of geologic materials using different instrument designs. We have employed a pulsed laser with both an ungated detector and a gated detector. A gated detector can reduce long-lived fluorescence while still collecting all Raman signal. In order to design a flight instrument, we need to quantify how natural surfaces will respond to laser stimulus. We define remote Raman efficiency of natural surfaces as the ratio of radiant exitance leaving a natural surface to the irradiance of the incident laser. The radiant exitance of a natural surface is the product of the sample radiance, the projected solid angle, and the full-width-half-maximum of the Raman signal. We have determined the remote Raman efficiency for a variety of rocks and minerals. The best efficiencies are achieved for large, clear, single crystals that produce the most radiant exitance, while darker fine-grained mineral mixtures produce lower efficiencies. By implementing a pulsed laser, gated detector system we have improved the signal detection and have generally decreased the integration time necessary to detect Raman signal from natural surfaces.

Characterization of retroviral infectivity and superinfection resistance during retrovirus-mediated transduction of mammalian cells.

PubMed

Liao, J; Wei, Q; Fan, J; Zou, Y; Song, D; Liu, J; Liu, F; Ma, C; Hu, X; Li, L; Yu, Y; Qu, X; Chen, L; Yu, X; Zhang, Z; Zhao, C; Zeng, Z; Zhang, R; Yan, S; Wu, T; Wu, X; Shu, Y; Lei, J; Li, Y; Zhang, W; Wang, J; Reid, R R; Lee, M J; Huang, W; Wolf, J M; He, T-C; Wang, J

2017-06-01

Retroviral vectors including lentiviral vectors are commonly used tools to stably express transgenes or RNA molecules in mammalian cells. Their utilities are roughly divided into two categories, stable overexpression of transgenes and RNA molecules, which requires maximal transduction efficiency, or functional selection with retrovirus (RV)-based libraries, which takes advantage of retroviral superinfection resistance. However, the dynamic features of RV-mediated transduction are not well characterized. Here, we engineered two murine stem cell virus-based retroviral vectors expressing dual fluorescence proteins and antibiotic markers, and analyzed virion production efficiency and virion stability, dynamic infectivity and superinfection resistance in different cell types, and strategies to improve transduction efficiency. We found that the highest virion production occurred between 60 and 72 h after transfection. The stability of the collected virion supernatant decreased by >60% after 3 days in storage. We found that RV infectivity varied drastically in the tested human cancer lines, while low transduction efficiency was partially overcome with increased virus titer, prolonged infection duration and/or repeated infections. Furthermore, we demonstrated that RV receptors PIT1 and PIT2 were lowly expressed in the analyzed cells, and that PIT1 and/or PIT2 overexpression significantly improved transduction efficiency in certain cell lines. Thus, our findings provide resourceful information for the optimal conditions of retroviral-mediated gene delivery.

The correlation of the maximum intensity of fluorescence with pigment characteristics of leaves of Betula pendula

NASA Astrophysics Data System (ADS)

Zavoruev, V. V.; Zavorueva, E. N.

2015-11-01

Using fluorimeter Junior PAM (Heinz Walz GmbH, Germany) the fluorescence parameters of leaves of Betula pendula are investigated. A linear dependence of the maximum fluorescence (Fm) of leaves from the ratio of total chlorophylls concentration to concentration of carotenoids is obtained. Such dependence is found for samples collected during the period of vegetation and for simultaneous selection of colored leaves.

Site-specific multipoint fluorescence measurement system with end-capped optical fibers.

PubMed

Song, Woosub; Moon, Sucbei; Lee, Byoung-Cheol; Park, Chul-Seung; Kim, Dug Young; Kwon, Hyuk Sang

2011-07-10

We present the development and implementation of a spatially and spectrally resolved multipoint fluorescence correlation spectroscopy (FCS) system utilizing multiple end-capped optical fibers and an inexpensive laser source. Specially prepared end-capped optical fibers placed in an image plane were used to both collect fluorescence signals from the sample and to deliver signals to the detectors. The placement of independently selected optical fibers on the image plane was done by monitoring the end-capped fiber tips at the focus using a CCD, and fluorescence from specific positions of a sample were collected by an end-capped fiber, which could accurately represent light intensities or spectral data without incurring any disturbance. A fast multipoint spectroscopy system with a time resolution of ∼1.5 ms was then implemented using a prism and an electron multiplying charge coupled device with a pixel binning for the region of interest. The accuracy of our proposed system was subsequently confirmed by experimental results, based on an FCS analysis of microspheres in distilled water. We expect that the proposed multipoint site-specific fluorescence measurement system can be used as an inexpensive fluorescence measurement tool to study many intracellular and molecular dynamics in cell biology. © 2011 Optical Society of America

One-step synthesis of fluorescein modified nano-carbon for Pd(II) detection via fluorescence quenching.

PubMed

Panchompoo, Janjira; Aldous, Leigh; Baker, Matthew; Wallace, Mark I; Compton, Richard G

2012-05-07

Carbon black (CB) nanoparticles modified with fluorescein, a highly fluorescent molecule, were prepared using a facile and efficient methodology. Simply stirring CB in aqueous solution containing fluorescein resulted in the strong physisorption of fluorescein onto the CB surface. The resulting Fluorescein/CB was then characterised by means of X-ray photoelectron spectroscopy (XPS), cyclic voltammetry (CV), fluorescence microscopy and fluorescence spectroscopy. The optimum experimental conditions for fluorescence of Fluorescein/CB viz. fluorescence excitation and emission wavelengths, O(2) removal and the amount of Fluorescein/CB used, were investigated. The Fluorescein/CB was used as a fluorescent probe for the sensitive detection of Pd(II) in water, based on fluorescence quenching. The results demonstrated that the fluorescence intensity of Fluorescein/CB decreased with increasing Pd(II) concentration, and the fluorescence quenching process could be described by the Stern-Volmer equation. The limit of detection (LOD) for the fluorescence quenching of Fluorescein/CB by Pd(II) in aqueous solution was found to be 1.07 μM (based on 3σ). Last, approaches were studied for the removal of Fe(III) which interferes with the fluorescence quenching of Fluorescein/CB. Complexation of Fe(III) with salicylic acid was used to enhance and control the selectivity of Fluorescein/CB sensor towards Pd(II) in the presence of Fe(III).

[Sensitive Determination of Chondroitin Sulfate by Fluorescence Recovery of an Anionic Aluminum Phthalocyanine-Cationic Surfactant Ion-Association Complex Used as a Fluorescent Probe Emitting at Red Region].

PubMed

Chen, Lin; Huang, Ping; Yang, Hui-qing; Deng, Ya-bin; Guo, Meng-lin; Li, Dong-hui

2015-08-01

Determination of chondroitin sulfate in the biomedical field has an important value. The conventional methods for the assay of chondroitin sulfate are still unsatisfactory in sensitivity, selectivity or simplicity. This work aimed at developing a novel method for sensitive and selective determination of chondroitin sulfate by fluorimetry. We found that some kinds of cationic surfactants have the ability to quench the fluorescence of tetrasulfonated aluminum phthalocyanine (AlS4Pc), a strongly fluorescent compound which emits at red region, with high efficiency. But, the fluorescence of the above-mentioned fluorescence quenching system recovered significantly when chondroitin sulfate (CS) exits. Tetradecyl dimethyl benzyl ammonium chloride(TDBAC) which was screened from all of the candidates of cationic surfactants was chosen as the quencher because it shows the most efficient quenching effect. It was found that the fluorescence of AlS4Pc was extremely quenched by TDBAC because of the formation of association complex between AlS4Pc and TDBAC. Fluorescence of the association complex recovered dramatically after the addition of chondroitin sulfate (CS) due to the ability of chondroitin sulfate to shift the association equilibrium of the association, leading to the release of AlS4Pc, thus resulting in an increase in the fluorescence of the reaction system. Based on this phenomenon, a novel method with simplicity, accuracy and sensitivity was developed for quantitative determination of CS. Factors including the reaction time, influencing factors and the effect of coexisting substances were investigated and discussed. Under optimum conditions the linear range of the calibration curve was 0.20~10.0 μg · mL(-1). The detection limit for CS was 0.070 μg · mL(-1). The method has been applied to the analysis of practical samples with satisfied results. This work expands the applications of AlS4Pc in biomedical area.

Design and field application of a UV-LED based optical fiber biofilm sensor.

PubMed

Fischer, Matthias; Wahl, Martin; Friedrichs, Gernot

2012-03-15

Detecting changes in the formation dynamics of biofilms stemming from bacteria and unicellular microorganisms in their natural environment is of prime interest for biological, ecological as well as anti-fouling technology research. We developed a robust optical fiber-based biofilm sensor ready to be applied in natural aquatic environments for on-line, in situ and non-destructive monitoring of large-area biofilms. The device is based on the detection of the natural fluorescence of microorganisms constituting the biofilm. Basically, the intrinsic fluorescence of the amino acid tryptophan is excited at a wavelength of λ=280 nm and detected at λ=350 nm utilising a numerically optimized sensor head equipped with a UV-LED light source and optical fiber bundles for efficient fluorescence light collection. Calibration was carried out with tryptophan solutions and two characteristic marine bacteria strains revealing linear signal response, satisfactory background suppression, wide dynamic range, and an experimental detection limit of 4 × 10(3)cells/cm(2). Successful field experiments in the Baltic Sea accomplished over a period of twenty-one days provided for the first time continuous observation of biofilm formation dynamics in a natural habitat. Starting from the first adhering bacteria, the measurement yielded the characteristic three phases of biofilm formation up to a fully developed biofilm. The sensor system holds potential for applications in aquatic sciences including deep sea research and, after further miniaturisation, in the industrial and biomedical field. Copyright © 2012 Elsevier B.V. All rights reserved.

A fluorescent-based HPLC assay for quantification of cysteine and cysteamine adducts in Escherichia coli-derived proteins.

PubMed

Soriano, Brian D; Tam, Lei-Ting T; Lu, Hsieng S; Valladares, Violeta G

2012-01-01

Recombinant proteins expressed in Escherichia coli are often produced as unfolded, inactive forms accumulated in inclusion bodies. Redox-coupled thiols are typically employed in the refolding process in order to catalyze the formation of correct disulfide bonds at maximal folding efficiency. These thiols and the recombinant proteins can form mixed disulfide bonds to generate thiol-protein adducts. In this work, we apply a fluorescent-based assay for the quantification of cysteine and cysteamine adducts as observed in E. coli-derived proteins. The thiols are released by reduction of the adducted protein, collected and labeled with a fluorescent reagent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. The derivatized thiols are separated by reversed-phase HPLC and can be accurately quantified after method optimization. The estimated thiol content represents total amount of adducted forms present in the analyzed samples. The limit of quantification (LOQ) was established; specifically, the lowest amount of quantifiable cysteine adduction is 30 picograms and the lowest amount of quantifiable cysteamine adduction is 60 picograms. The assay is useful for quantification of adducts in final purified products as well as in-process samples from various purification steps. The assay indicates that the purification process accomplishes a decrease in cysteine adduction from 0.19 nmol adduct/nmol protein to 0.03 nmol adduct/nmol protein as well as a decrease in cysteamine adduction from 0.24 nmol adduct/nmol protein to 0.14 nmol adduct/nmol protein. Copyright © 2011. Published by Elsevier B.V.

Fluorescence Live Cell Imaging

PubMed Central

Ettinger, Andreas

2014-01-01

Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio, and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities, and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate fluorescent protein constructs by spinning disk confocal microscopy. PMID:24974023

Highly Efficient Nondoped Green Organic Light-Emitting Diodes with Combination of High Photoluminescence and High Exciton Utilization.

PubMed

Wang, Chu; Li, Xianglong; Pan, Yuyu; Zhang, Shitong; Yao, Liang; Bai, Qing; Li, Weijun; Lu, Ping; Yang, Bing; Su, Shijian; Ma, Yuguang

2016-02-10

Photoluminescence (PL) efficiency and exciton utilization efficiency are two key parameters to harvest high-efficiency electroluminescence (EL) in organic light-emitting diodes (OLEDs). But it is not easy to simultaneously combine these two characteristics (high PL efficiency and high exciton utilization) into a fluorescent material. In this work, an efficient combination was achieved through two concepts of hybridized local and charge-transfer (CT) state (HLCT) and "hot exciton", in which the former is responsible for high PL efficiency while the latter contributes to high exciton utilization. On the basis of a tiny chemical modification in TPA-BZP, a green-light donor-acceptor molecule, we designed and synthesized CzP-BZP with this efficeient combination of high PL efficiency of η(PL) = 75% in the solid state and maximal exciton utilization efficiency up to 48% (especially, the internal quantum efficiency of η(IQE) = 35% substantially exceed 25% of spin statistics limit) in OLED. The nondoped OLED of CzP-BZP exhibited an excellent performance: a green emission with a CIE coordinate of (0.34, 0.60), a maximum current efficiency of 23.99 cd A(-1), and a maximum external quantum efficiency (EQE, η(EQE)) of 6.95%. This combined HLCT state and "hot exciton" strategy should be a practical way to design next-generation, low-cost, high-efficiency fluorescent OLED materials.

5-ALA-induced fluorescence as a marker for diagnostic tissue in stereotactic biopsies of intracranial lymphomas: experience in 41 patients.

PubMed

Kiesel, Barbara; Millesi, Matthias; Woehrer, Adelheid; Furtner, Julia; Bavand, Anahita; Roetzer, Thomas; Mischkulnig, Mario; Wolfsberger, Stefan; Preusser, Matthias; Knosp, Engelbert; Widhalm, Georg

2018-06-01

OBJECTIVE Stereotactic needle biopsies are usually performed for histopathological confirmation of intracranial lymphomas to guide adequate treatment. During biopsy, intraoperative histopathology is an effective tool to avoid acquisition of nondiagnostic samples. In the last years, 5-aminolevulinic acid (5-ALA)-induced fluorescence has been increasingly used for visualization of diagnostic brain tumor tissue during stereotactic biopsies. Recently, visible fluorescence was reported in the first cases of intracranial lymphomas as well. The aim of this study is thus to investigate the technical and clinical utility of 5-ALA-induced fluorescence in a large series of stereotactic biopsies for intracranial lymphoma. METHODS This prospective study recruited adult patients who underwent frameless stereotactic needle biopsy for a radiologically suspected intracranial lymphoma after oral 5-ALA administration. During biopsy, samples from the tumor region were collected for histopathological analysis, and presence of fluorescence (strong, vague, or no fluorescence) was assessed with a modified neurosurgical microscope. In tumors with available biopsy samples from at least 2 different regions the intratumoral fluorescence homogeneity was additionally investigated. Furthermore, the influence of potential preoperative corticosteroid treatment or immunosuppression on fluorescence was analyzed. Histopathological tumor diagnosis was established and all collected biopsy samples were screened for diagnostic lymphoma tissue. RESULTS The final study cohort included 41 patients with intracranial lymphoma. Stereotactic biopsies with assistance of 5-ALA were technically feasible in all cases. Strong fluorescence was found as maximum level in 30 patients (75%), vague fluorescence in 2 patients (4%), and no visible fluorescence in 9 patients (21%). In 28 cases, samples were obtained from at least 2 different tumor regions; homogenous intratumoral fluorescence was found in 16 of those cases (57%) and inhomogeneous intratumoral fluorescence in 12 (43%). According to histopathological analysis, all samples with strong or vague fluorescence contained diagnostic lymphoma tissue, resulting in a positive predictive value of 100%. Analysis showed no influence of preoperative corticosteroids or immunosuppression on fluorescence. CONCLUSIONS The data obtained in this study demonstrate the technical and clinical utility of 5-ALA-induced fluorescence in stereotactic biopsies of intracranial lymphomas. Thus, 5-ALA can serve as a useful tool to select patients not requiring intraoperative histopathology, and its application should markedly reduce operation time and related costs in the future.

Optical radiation emissions from compact fluorescent lamps.

PubMed

Khazova, M; O'Hagan, J B

2008-01-01

There is a drive to energy efficiency to mitigate climate change. To meet this challenge, the UK Government has proposed phasing out incandescent lamps by the end of 2011 and replacing them with energy efficient fluorescent lighting, including compact fluorescent lamps (CFLs) with integrated ballasts. This paper presents a summary of an assessment conducted by the Health Protection Agency in March 2008 to evaluate the optical radiation emissions of CFLs currently available in the UK consumer market. The study concluded that the UV emissions from a significant percentage of the tested CFLs with single envelopes may result in foreseeable overexposure of the skin when these lamps are used in desk or task lighting applications. The optical output of all tested CFLs, in addition to high-frequency modulation, had a 100-Hz envelope with modulation in excess of 15%. This degree of modulation may be linked to a number of adverse effects.

Influence of silver nanoparticles on relaxation processes and efficiency of dipole – dipole energy transfer between dye molecules in polymethylmethacrylate films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bryukhanov, V V; Borkunov, R Yu; Tsarkov, M V

The fluorescence and phosphorescence of dyes in thin polymethylmethacrylate (PMMA) films in the presence of ablated silver nanoparticles has been investigated in a wide temperature range by methods of femtosecond and picosecond laser photoexcitation. The fluorescence and phosphorescence times, as well as spectral and kinetic characteristics of rhodamine 6G (R6G) molecules in PMMA films are measured in a temperature range of 80 – 330 K. The temperature quenching activation energy of the fluorescence of R6G molecules in the presence of ablated silver nanoparticles is found. The vibrational relaxation rate of R6G in PMMA films is estimated, the efficiency of themore » dipole – dipole electron energy transfer between R6G and brilliant green molecules (enhanced by plasmonic interaction with ablated silver nanoparticles) is analysed, and the constants of this energy transfer are determined. (nanophotonics)« less

A novel broad specificity fucosidase capable of core α1-6 fucose release from N-glycans labeled with urea-linked fluorescent dyes.

PubMed

Vainauskas, Saulius; Kirk, Charlotte H; Petralia, Laudine; Guthrie, Ellen P; McLeod, Elizabeth; Bielik, Alicia; Luebbers, Alex; Foster, Jeremy M; Hokke, Cornelis H; Rudd, Pauline M; Shi, Xiaofeng; Taron, Christopher H

2018-06-22

Exoglycosidases are often used for detailed characterization of glycan structures. Bovine kidney α-fucosidase is commonly used to determine the presence of core α1-6 fucose on N-glycans, an important modification of glycoproteins. Recently, several studies have reported that removal of core α1-6-linked fucose from N-glycans labeled with the reactive N-hydroxysuccinimide carbamate fluorescent labels 6-aminoquinolyl-N-hydroxysuccinimidylcarbamate (AQC) and RapiFluor-MS is severely impeded. We report here the cloning, expression and biochemical characterization of an α-fucosidase from Omnitrophica bacterium (termed fucosidase O). We show that fucosidase O can efficiently remove α1-6- and α1-3-linked core fucose from N-glycans. Additionally, we demonstrate that fucosidase O is able to efficiently hydrolyze core α1-6-linked fucose from N-glycans labeled with any of the existing NHS-carbamate activated fluorescent dyes.

A comparative study of fluorescent and LED lighting in industrial facilities

NASA Astrophysics Data System (ADS)

Perdahci PhD, C.; Akin BSc, H. C.; Cekic Msc, O.

2018-05-01

Industrial facilities have always been in search for reducing outgoings and minimizing energy consumption. Rapid developments in lighting technology require more energy efficient solutions not only for industries but also for many sectors and for households. Addition of solid-state technology has brought LED lamps into play and with LED lamp usage, efficacy level has reached its current values. Lighting systems which uses fluorescent and LED lamps have become the prior choice for many industrial facilities. This paper presents a comparative study about fluorescent and LED based indoor lighting systems for a warehouse building in an industrial facility in terms of lighting distribution values, colour rendering, power consumption, energy efficiency and visual comfort. Both scenarios have been modelled and simulated by using Relux and photometric data for the luminaires have been gathered by conducting tests and measurements in an accredited laboratory.

Triazole-based Zn²⁺-specific molecular marker for fluorescence bioimaging.

PubMed

Sinha, Sougata; Mukherjee, Trinetra; Mathew, Jomon; Mukhopadhyay, Subhra K; Ghosh, Subrata

2014-04-25

Fluorescence bioimaging potential, both in vitro and in vivo, of a yellow emissive triazole-based molecular marker has been investigated and demonstrated. Three different kinds of cells, viz Bacillus thuringiensis, Candida albicans, and Techoma stans pollen grains were used to investigate the intracellular zinc imaging potential of 1 (in vitro studies). Fluorescence imaging of translocation of zinc through the stem of small herb, Peperomia pellucida, having transparent stem proved in vivo bioimaging capability of 1. This approach will enable in screening cell permeability and biostability of a newly developed probe. Similarly, the current method for detection and localization of zinc in Gram seed sprouts could be an easy and potential alternative of the existing analytical methods to investigate the efficiency of various strategies applied for increasing zinc-content in cereal crops. The probe-zinc ensemble has efficiently been applied for detecting phosphate-based biomolecules. Copyright © 2014 Elsevier B.V. All rights reserved.

Oligothiophene-based colorimetric and ratiometric fluorescence dual-channel cyanide chemosensor: Sensing ability, TD-DFT calculations and its application as an efficient solid state sensor

NASA Astrophysics Data System (ADS)

Lan, Linxin; Li, Tianduo; Wei, Tao; Pang, He; Sun, Tao; Wang, Enhua; Liu, Haixia; Niu, Qingfen

2018-03-01

An oligothiophene-based colorimetric and ratiometric fluorescence dual-channel cyanide chemosensor 3 T-2CN was reported. Sensor 3 T-2CN showed both naked-eye recognition and ratiometric fluorescence response for CN- with an excellent selectivity and high sensitivity. The sensing mechanism based on the nucleophilic attack of CN- on the vinyl Cdbnd C bond has been successfully confirmed by the optical measurements, 1H NMR titration, FT-IR spectra as well as the DFT/TD-DFT calculations. Moreover, the detection limit was calculated to be 0.19 μM, which is much lower than the maximum permission concentration in drinking water (1.9 μM). Importantly, test strips (filter paper and TLC plates) containing 3 T-2CN were fabricated, which could act as a practical and efficient solid state optical sensor for CN- in field measurements.

Nanocapillaries for Open Tubular Chromatographic Separations of Proteins in Femtoliter to Picoliter Samples

PubMed Central

Wang, Xiayan; Cheng, Chang; Wang, Shili; Zhao, Meiping; Dasgupta, Purnendu K.; Liu, Shaorong

2009-01-01

We have recently examined the potential of bare nanocapillaries for free solution DNA separations and demonstrated efficiencies exceeding 106 theoretical plates/m. In the present work, we demonstrate the use of bare and hydroxypropylcellulose (HPC) coated open tubular nanocapillaries for protein separations. Using 1.5 μm inner diameter (i.d.) capillary columns, hydrodynamically injecting femto to picoliter (fL-pL) volumes of fluorescent or fluorescent dye labeled protein samples, utilizing a pneumatically pressurized chamber containing 1.0 mM sodium tetraborate solution eluent (typ. 200 psi) as the pump and performing on-column detection using a simple laser-induced fluorescence detector, we demonstrate efficiencies of close to a million theoretical plates/m while generating single digit μL volumes of waste for a complete chromatographic run. We achieve baseline resolution for a protein mixture consisting of transferrin, α-lactalbumin, insulin, and α -2-macroglobulin. PMID:19663450

2D fluorescence spectroscopy for monitoring ion-exchange membrane based technologies - Reverse electrodialysis (RED).

PubMed

Pawlowski, Sylwin; Galinha, Claudia F; Crespo, João G; Velizarov, Svetlozar

2016-01-01

Reverse electrodialysis (RED) is one of the emerging, membrane-based technologies for harvesting salinity gradient energy. In RED process, fouling is an undesirable operation constraint since it leads to a decrease of the obtainable net power density due to increasing stack electric resistance and pressure drop. Therefore, early fouling detection is one of the main challenges for successful RED technology implementation. In the present study, two-dimensional (2D) fluorescence spectroscopy was used, for the first time, as a tool for fouling monitoring in RED. Fluorescence excitation-emission matrices (EEMs) of ion-exchange membrane surfaces and of natural aqueous streams were acquired during one month of a RED stack operation. Fouling evolvement on the ion-exchange membrane surfaces was successfully followed by 2D fluorescence spectroscopy and quantified using principal components analysis (PCA). Additionally, the efficiency of cleaning strategy was assessed by measuring the membrane fluorescence emission intensity before and after cleaning. The anion-exchange membrane (AEM) surface in contact with river water showed to be significantly affected due to fouling by humic compounds, which were found to cross through the membrane from the lower salinity (river water) to higher salinity (sea water) stream. The results obtained show that the combined approach of using 2D fluorescence spectroscopy and PCA has a high potential for studying fouling development and membrane cleaning efficiency in ion exchange membrane processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Development and biodistribution of a theranostic aluminum phthalocyanine nanophotosensitizer.

PubMed

Asem, Heba; El-Fattah, Ahmed Abd; Nafee, Noha; Zhao, Ying; Khalil, Labiba; Muhammed, Mamoun; Hassan, Moustapha; Kandil, Sherif

2016-03-01

Aluminum phthalocyanine (AlPc) is an efficient second generation photosensitizer (PS) with high fluorescence ability. Its use in photodynamic therapy (PDT) is hampered by hydrophobicity and poor biodistribution. AlPc was converted to a biocompatible nanostructure by incorporation into amphiphilic polyethylene glycol-polycaprolactone (PECL) copolymer nanoparticles, allowing efficient entrapment of the PS in the hydrophobic core, water dispersibility and biodistribution enhancement by PEG-induced surface characteristics. A series of synthesized PECL copolymers were used to prepare nanophotosensitizers with an average diameter of 66.5-99.1nm and encapsulation efficiency (EE%) of 66.4-78.0%. One formulation with favorable colloidal properties and relatively slow release over 7 days was selected for in vitro photophysical assessment and in vivo biodistribution studies in mice. The photophysical properties of AlPc were improved by encapsulating AlPc into PECL-NPs, which showed intense fluorescence emission at 687nm and no AlPc aggregation has been induced after entrapment into the nanoparticles. Biodistribution of AlPc loaded NPs (AlPc-NPs) and free AlPc drug in mice was monitored by in vivo whole body fluorescence imaging and ex vivo organ imaging, with in vivo imaging system (IVIS). Compared to a AlPc solution in aqueous TWEEN 80 (2 w/v%), the developed nanophotosensitizer showed targeted drug delivery to lungs, liver and spleen as monitored by the intrinsic fluorescence of AlPc at different time points (1h, 24h and 48h) post iv. administration. The AlPc-based copolymer nanoparticles developed offer potential as a single agent-multifunctional theranostic nanophotosensitizer for PDT coupled with imaging-guided drug delivery and biodistribution, and possibly also fluorescence diagnostics. Copyright © 2015 Elsevier B.V. All rights reserved.

5-Aminolevulinic Acid Induced Fluorescence Is a Powerful Intraoperative Marker for Precise Histopathological Grading of Gliomas with Non-Significant Contrast-Enhancement

PubMed Central

Widhalm, Georg; Kiesel, Barbara; Woehrer, Adelheid; Traub-Weidinger, Tatjana; Preusser, Matthias; Marosi, Christine; Prayer, Daniela; Hainfellner, Johannes A.; Knosp, Engelbert; Wolfsberger, Stefan

2013-01-01

Background Intraoperative identification of anaplastic foci in diffusely infiltrating gliomas (DIG) with non-significant contrast-enhancement on MRI is indispensible to avoid histopathological undergrading and subsequent treatment failure. Recently, we found that 5-aminolevulinic acid (5-ALA) induced protoporphyrin IX (PpIX) fluorescence can visualize areas with increased proliferative and metabolic activity in such gliomas intraoperatively. As treatment of DIG is predominantely based on histopathological World Health Organisation (WHO) parameters, we analyzed whether PpIX fluorescence can detect anaplastic foci according to these criteria. Methods We prospectively included DIG patients with non-significant contrast-enhancement that received 5-ALA prior to resection. Intraoperatively, multiple samples from PpIX positive and negative intratumoral areas were collected using a modified neurosurgical microscope. In all samples, histopathological WHO criteria and proliferation rate were assessed and correlated to the PpIX fluorescence status. Results A total of 215 tumor specimens were collected in 59 patients. Of 26 WHO grade III gliomas, 23 cases (85%) showed focal PpIX fluorescence, whereas 29 (91%) of 33 WHO grade II gliomas were PpIX negative. In intratumoral areas with focal PpIX fluorescence, mitotic rate, cell density, nuclear pleomorphism, and proliferation rate were significantly higher than in non-fluorescing areas. The positive predictive value of focal PpIX fluorescence for WHO grade III histology was 85%. Conclusions Our study indicates that 5-ALA induced PpIX fluorescence is a powerful marker for intraoperative identification of anaplastic foci according to the histopathological WHO criteria in DIG with non-significant contrast-enhancement. Therefore, application of 5-ALA optimizes tissue sampling for precise histopathological diagnosis independent of brain-shift. PMID:24204718

5-Aminolevulinic acid induced fluorescence is a powerful intraoperative marker for precise histopathological grading of gliomas with non-significant contrast-enhancement.

PubMed

Widhalm, Georg; Kiesel, Barbara; Woehrer, Adelheid; Traub-Weidinger, Tatjana; Preusser, Matthias; Marosi, Christine; Prayer, Daniela; Hainfellner, Johannes A; Knosp, Engelbert; Wolfsberger, Stefan

2013-01-01

Intraoperative identification of anaplastic foci in diffusely infiltrating gliomas (DIG) with non-significant contrast-enhancement on MRI is indispensible to avoid histopathological undergrading and subsequent treatment failure. Recently, we found that 5-aminolevulinic acid (5-ALA) induced protoporphyrin IX (PpIX) fluorescence can visualize areas with increased proliferative and metabolic activity in such gliomas intraoperatively. As treatment of DIG is predominantely based on histopathological World Health Organisation (WHO) parameters, we analyzed whether PpIX fluorescence can detect anaplastic foci according to these criteria. We prospectively included DIG patients with non-significant contrast-enhancement that received 5-ALA prior to resection. Intraoperatively, multiple samples from PpIX positive and negative intratumoral areas were collected using a modified neurosurgical microscope. In all samples, histopathological WHO criteria and proliferation rate were assessed and correlated to the PpIX fluorescence status. A total of 215 tumor specimens were collected in 59 patients. Of 26 WHO grade III gliomas, 23 cases (85%) showed focal PpIX fluorescence, whereas 29 (91%) of 33 WHO grade II gliomas were PpIX negative. In intratumoral areas with focal PpIX fluorescence, mitotic rate, cell density, nuclear pleomorphism, and proliferation rate were significantly higher than in non-fluorescing areas. The positive predictive value of focal PpIX fluorescence for WHO grade III histology was 85%. Our study indicates that 5-ALA induced PpIX fluorescence is a powerful marker for intraoperative identification of anaplastic foci according to the histopathological WHO criteria in DIG with non-significant contrast-enhancement. Therefore, application of 5-ALA optimizes tissue sampling for precise histopathological diagnosis independent of brain-shift.

ultraLM and miniLM: Locator tools for smart tracking of fluorescent cells in correlative light and electron microscopy.

PubMed

Brama, Elisabeth; Peddie, Christopher J; Wilkes, Gary; Gu, Yan; Collinson, Lucy M; Jones, Martin L

2016-12-13

In-resin fluorescence (IRF) protocols preserve fluorescent proteins in resin-embedded cells and tissues for correlative light and electron microscopy, aiding interpretation of macromolecular function within the complex cellular landscape. Dual-contrast IRF samples can be imaged in separate fluorescence and electron microscopes, or in dual-modality integrated microscopes for high resolution correlation of fluorophore to organelle. IRF samples also offer a unique opportunity to automate correlative imaging workflows. Here we present two new locator tools for finding and following fluorescent cells in IRF blocks, enabling future automation of correlative imaging. The ultraLM is a fluorescence microscope that integrates with an ultramicrotome, which enables 'smart collection' of ultrathin sections containing fluorescent cells or tissues for subsequent transmission electron microscopy or array tomography. The miniLM is a fluorescence microscope that integrates with serial block face scanning electron microscopes, which enables 'smart tracking' of fluorescent structures during automated serial electron image acquisition from large cell and tissue volumes.

Video-Rate Confocal Microscopy for Single-Molecule Imaging in Live Cells and Superresolution Fluorescence Imaging

PubMed Central

Lee, Jinwoo; Miyanaga, Yukihiro; Ueda, Masahiro; Hohng, Sungchul

2012-01-01

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0–85 μm from the surface of a coverglass. PMID:23083712

Wide field fluorescence epi-microscopy behind a scattering medium enabled by speckle correlations

NASA Astrophysics Data System (ADS)

Hofer, Matthias; Soeller, Christian; Brasselet, Sophie; Bertolotti, Jacopo

2018-04-01

Fluorescence microscopy is widely used in biological imaging, however scattering from tissues strongly limits its applicability to a shallow depth. In this work we adapt a methodology inspired from stellar speckle interferometry, and exploit the optical memory effect to enable fluorescence microscopy through a turbid layer. We demonstrate efficient reconstruction of micrometer-size fluorescent objects behind a scattering medium in epi-microscopy, and study the specificities of this imaging modality (magnification, field of view, resolution) as compared to traditional microscopy. Using a modified phase retrieval algorithm to reconstruct fluorescent objects from speckle images, we demonstrate robust reconstructions even in relatively low signal to noise conditions. This modality is particularly appropriate for imaging in biological media, which are known to exhibit relatively large optical memory ranges compatible with tens of micrometers size field of views, and large spectral bandwidths compatible with emission fluorescence spectra of tens of nanometers widths.

An efficient method for visualization and growth of fluorescent Xanthomonas oryzae pv. oryzae in planta

PubMed Central

Han, Sang-Wook; Park, Chang-Jin; Lee, Sang-Won; Ronald, Pamela C

2008-01-01

Background Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight disease, is a serious pathogen of rice. Here we describe a fluorescent marker system to study virulence and pathogenicity of X. oryzae pv. oryzae. Results A fluorescent X. oryzae pv. oryzae Philippine race 6 strain expressing green fluorescent protein (GFP) (PXO99GFP) was generated using the gfp gene under the control of the neomycin promoter in the vector, pPneo-gfp. The PXO99GFPstrain displayed identical virulence and avirulence properties as the wild type control strain, PXO99. Using fluorescent microscopy, bacterial multiplication and colonization were directly observed in rice xylem vessels. Accurate and rapid determination of bacterial growth was assessed using fluoremetry and an Enzyme-Linked ImmunoSorbant Assay (ELISA). Conclusion Our results indicate that the fluorescent marker system is useful for assessing bacterial infection and monitoring bacterial multiplication in planta. PMID:18826644

Shrink-induced silica multiscale structures for enhanced fluorescence from DNA microarrays.

PubMed

Sharma, Himanshu; Wood, Jennifer B; Lin, Sophia; Corn, Robert M; Khine, Michelle

2014-09-23

We describe a manufacturable and scalable method for fabrication of multiscale wrinkled silica (SiO2) structures on shrink-wrap film to enhance fluorescence signals in DNA fluorescence microarrays. We are able to enhance the fluorescence signal of hybridized DNA by more than 120 fold relative to a planar glass slide. Notably, our substrate has improved detection sensitivity (280 pM) relative to planar glass slide (11 nM). Furthermore, this is accompanied by a 30-45 times improvement in the signal-to-noise ratio (SNR). Unlike metal enhanced fluorescence (MEF) based enhancements, this is a far-field and uniform effect based on surface concentration and photophysical effects from the nano- to microscale SiO2 structures. Notably, the photophysical effects contribute an almost 2.5 fold enhancement over the concentration effects alone. Therefore, this simple and robust method offers an efficient technique to enhance the detection capabilities of fluorescence based DNA microarrays.

Shrink-Induced Silica Multiscale Structures for Enhanced Fluorescence from DNA Microarrays

PubMed Central

2015-01-01

We describe a manufacturable and scalable method for fabrication of multiscale wrinkled silica (SiO2) structures on shrink-wrap film to enhance fluorescence signals in DNA fluorescence microarrays. We are able to enhance the fluorescence signal of hybridized DNA by more than 120 fold relative to a planar glass slide. Notably, our substrate has improved detection sensitivity (280 pM) relative to planar glass slide (11 nM). Furthermore, this is accompanied by a 30–45 times improvement in the signal-to-noise ratio (SNR). Unlike metal enhanced fluorescence (MEF) based enhancements, this is a far-field and uniform effect based on surface concentration and photophysical effects from the nano- to microscale SiO2 structures. Notably, the photophysical effects contribute an almost 2.5 fold enhancement over the concentration effects alone. Therefore, this simple and robust method offers an efficient technique to enhance the detection capabilities of fluorescence based DNA microarrays. PMID:25191785

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

PubMed

Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

2015-12-14

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

Optical properties of flexible fluorescent films prepared by screen printing technology

NASA Astrophysics Data System (ADS)

Chen, Yan; Ke, Taiyan; Chen, Shuijin; He, Xin; Zhang, Mei; Li, Dong; Deng, Jinfeng; Zeng, Qingguang

2018-05-01

In this work, we prepared a fluorescent film comprised phosphors and silicone on flexible polyethylene terephthalate (PET) substrate using a screen printing technology. The effects of mesh number and weight ratio of phosphors to silicone on the optical properties of the flexible films were investigated. The results indicate that the emission intensity of the film increase as the mesh decreased from 400 to 200, but the film surface gradually becomes uneven. The fluorescent film with high emission intensity and smooth surface can be obtained when the weight ratio of phosphor to gel is 2:1, and mesh number is 300. The luminous efficiency of the fabricated LEDs combined the fluorescent films with 460 nm Ga(In)N chip module can reach 75 lm/W. The investigation indicates that the approach can be applied in the remote fluorescent film conversion and decreases the requirements of the particle size and the dispersion state of fluorescent materials.

Structural basis for activity of highly efficient RNA mimics of green fluorescent protein

PubMed Central

Warner, Katherine Deigan; Chen, Michael C.; Song, Wenjiao; Strack, Rita L.; Thorn, Andrea; Jaffrey, Samie R.; Ferré-D’Amaré, Adrian R.

2014-01-01

Green fluorescent protein (GFP) and its derivatives revolutionized the study of proteins. Spinach is a recently reported in vitro evolved RNA mimic of GFP, which as genetically encoded fusions, makes possible live-cell, real-time imaging of biological RNAs, without resorting to large RNA-binding protein-GFP fusions. To elucidate the molecular basis of Spinach fluorescence, we have solved its co-crystal structure bound to its cognate exogenous chromophore, revealing that Spinach activates the small molecule by immobilizing it between a base triple, a G-quadruplex, and an unpaired guanine. Mutational and NMR analyses indicate that the G-quadruplex is essential for Spinach fluorescence, is also present in other fluorogenic RNAs, and may represent a general strategy for RNAs to induce fluorescence of chromophores. The structure has guided the design of a miniaturized 'Baby Spinach', and provides the foundation for structure-driven design and tuning of fluorescent RNAs. PMID:25026079

φ(2)GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples.

PubMed

Jain, Paras; Hartman, Travis E; Eisenberg, Nell; O'Donnell, Max R; Kriakov, Jordan; Govender, Karnishree; Makume, Mantha; Thaler, David S; Hatfull, Graham F; Sturm, A Willem; Larsen, Michelle H; Moodley, Preshnie; Jacobs, William R

2012-04-01

The difficulty of diagnosing active tuberculosis (TB) and lack of rapid drug susceptibility testing (DST) at the point of care remain critical obstacles to TB control. This report describes a high-intensity mycobacterium-specific-fluorophage (φ(2)GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing φ(2)GFP10 from previous reporter phages include an improved vector backbone with increased cloning capacity and superior expression of fluorescent reporter genes through use of an efficient phage promoter. φ(2)GFP10 produces a 100-fold increase in fluorescence per cell compared to existing reporter phages. DST for isoniazid and oxofloxacin, carried out in cultured samples, was complete within 36 h. Use of φ(2)GFP10 detected M. tuberculosis in clinical sputum samples collected from TB patients. DST for rifampin and kanamycin from sputum samples yielded results after 12 h of incubation with φ(2)GFP10. Fluorophage φ(2)GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection and for rapid DST.

Novel KCNQ2 channel activators discovered using fluorescence-based and automated patch-clamp-based high-throughput screening techniques

PubMed Central

Yue, Jin-feng; Qiao, Guan-hua; Liu, Ni; Nan, Fa-jun; Gao, Zhao-bing

2016-01-01

Aim: To establish an improved, high-throughput screening techniques for identifying novel KCNQ2 channel activators. Methods: KCNQ2 channels were stably expressed in CHO cells (KCNQ2 cells). Thallium flux assay was used for primary screening, and 384-well automated patch-clamp IonWorks Barracuda was used for hit validation. Two validated activators were characterized using a conventional patch-clamp recording technique. Results: From a collection of 80 000 compounds, the primary screening revealed a total of 565 compounds that potentiated the fluorescence signals in thallium flux assay by more than 150%. When the 565 hits were examined in IonWorks Barracuda, 38 compounds significantly enhanced the outward currents recorded in KCNQ2 cells, and were confirmed as KCNQ2 activators. In the conventional patch-clamp recordings, two validated activators ZG1732 and ZG2083 enhanced KCNQ2 currents with EC50 values of 1.04±0.18 μmol/L and 1.37±0.06 μmol/L, respectively. Conclusion: The combination of thallium flux assay and IonWorks Barracuda assay is an efficient high-throughput screening (HTS) route for discovering KCNQ2 activators. PMID:26725738

In vivo imaging of small animals with optical tomography and near-infrared fluorescent probes

NASA Astrophysics Data System (ADS)

Palmer, Matthew R.; Shibata, Yasushi; Kruskal, Jonathan B.; Lenkinski, Robert E.

2002-06-01

A developmental optical tomography has been designed for imaging small animals in vivo using near IR fluorophores. The system employs epi-illumination via a 450 W Xe arc lamp, filtered and collimated to illuminate a 10 cm square movable stage. Emission light is filtered then collected by a high- resolution, high quantum efficiency, cooled CCD camera. Stage movement and image acquisition are under the control of a personal computer running system integration and automation software. During an experiment, the anesthetized animal is secured to the stage and up to 200 projections can be acquired over 180 degrees rotation. Angular sampling of the light distribution at a point on the surface is used to determine relative contributions form ballistic and diffuse photons. We have employed the system to investigate a number of applications of in-vivo fluorescent imaging. In dynamic studies, hepatic function has been visualized in nude mice following intravenous injection of indocyanine green (ICG) and cerebrospinal fluid flow as been measured by injection of ICG-lipoprotein conjugate in the subarachnoid space of the lumbar spine followed by dynamic imaging of the brain. Further applications in physiological imaging, cancer detection, and molecular imaging are under investigation in our laboratory.

Triplet exciton confinement for enhanced fluorescent organic light-emitting diodes using a co-host system

NASA Astrophysics Data System (ADS)

Yoo, Han Kyu; Lee, Ho Won; Lee, Song Eun; Kim, Young Kwan; Kim, Se Hyun; Yoon, Seung Soo; Park, Jaehoon

2016-05-01

In this work, the co-host system within an emitting layer (EML) consists of the host and triplet managing (TM) host materials. A set of EML structures was fabricated with various concentrations of the TM host (0, 10, 30, 50, and 70%). The TM host triplet energy level is lower than the energy levels of the host and the guest, which leads to a reduction in the triplet exciton density and the singlet-triplet annihilation of the guest. Blue fluorescent organic light-emitting diodes exhibit a maximum luminous efficiency (LE) and an external quantum efficiency (EQE) of 9.74 cd/A and 4.92%, respectively. In addition, the efficiency roll-off ratios of the LE and the EQE are 14.25 and 13.16%, respectively.

Manganese Deficiency Leads to Genotype-Specific Changes in Fluorescence Induction Kinetics and State Transitions1[C][OA

PubMed Central

Husted, Søren; Laursen, Kristian H.; Hebbern, Christopher A.; Schmidt, Sidsel B.; Pedas, Pai; Haldrup, Anna; Jensen, Poul E.

2009-01-01

Barley (Hordeum vulgare) genotypes display a marked difference in their ability to tolerate growth at low manganese (Mn) concentrations, a phenomenon designated as differential Mn efficiency. Induction of Mn deficiency in two genotypes differing in Mn efficiency led to a decline in the quantum yield efficiency for both, although faster in the Mn-inefficient genotype. Leaf tissue and thylakoid Mn concentrations were reduced under Mn deficiency, but no difference between genotypes was observed and no visual Mn deficiency symptoms were developed. Analysis of the fluorescence induction kinetics revealed that in addition to the usual O-J-I-P steps, clear K and D steps were developed in the Mn-inefficient genotype under Mn deficiency. These marked changes indicated damages to photosystem II (PSII). This was further substantiated by state transition measurements, indicating that the ability of plants to redistribute excitation energy was reduced. The percentage change in state transitions for control plants with normal Mn supply of both genotypes was 9% to 11%. However, in Mn-deficient leaves of the Mn-inefficient genotypes, state transitions were reduced to less than 1%, whereas no change was observed for the Mn-efficient genotypes. Immunoblotting and the chlorophyll a/b ratio confirmed that Mn deficiency in general resulted in a significant reduction in abundance of PSII reaction centers relative to the peripheral antenna. In addition, PSII appeared to be significantly more affected by Mn limitation than PSI. However, the striking genotypic differences observed in Mn-deficient plants, when analyzing state transitions and fluorescence induction kinetics, could not be correlated with specific changes in photosystem proteins. Thus, there is no simple linkage between protein expression and the differential reduction in state transition and fluorescence induction kinetics observed for the genotypes under Mn deficiency. PMID:19369593

Experimental verification of the kinetic theory of FRET using optical microspectroscopy and obligate oligomers.

PubMed

Patowary, Suparna; Pisterzi, Luca F; Biener, Gabriel; Holz, Jessica D; Oliver, Julie A; Wells, James W; Raicu, Valerică

2015-04-07

Förster resonance energy transfer (FRET) is a nonradiative process for the transfer of energy from an optically excited donor molecule (D) to an acceptor molecule (A) in the ground state. The underlying theory predicting the dependence of the FRET efficiency on the sixth power of the distance between D and A has stood the test of time. In contrast, a comprehensive kinetic-based theory developed recently for FRET efficiencies among multiple donors and acceptors in multimeric arrays has waited for further testing. That theory has been tested in the work described in this article using linked fluorescent proteins located in the cytoplasm and at the plasma membrane of living cells. The cytoplasmic constructs were fused combinations of Cerulean as donor (D), Venus as acceptor (A), and a photo-insensitive molecule (Amber) as a nonfluorescent (N) place holder: namely, NDAN, NDNA, and ADNN duplexes, and the fully fluorescent quadruplex ADAA. The membrane-bound constructs were fused combinations of GFP2 as donor (D) and eYFP as acceptor (A): namely, two fluorescent duplexes (i.e., DA and AD) and a fluorescent triplex (ADA). According to the theory, the FRET efficiency of a multiplex such as ADAA or ADA can be predicted from that of analogs containing a single acceptor (e.g., NDAN, NDNA, and ADNN, or DA and AD, respectively). Relatively small but statistically significant differences were observed between the measured and predicted FRET efficiencies of the two multiplexes. While elucidation of the cause of this mismatch could be a worthy endeavor, the discrepancy does not appear to question the theoretical underpinnings of a large family of FRET-based methods for determining the stoichiometry and quaternary structure of complexes of macromolecules in living cells. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Efficient triplet application in exciplex delayed-fluorescence OLEDs using a reverse intersystem crossing mechanism based on a ΔES-T of around zero.

PubMed

Zhang, Tianyou; Chu, Bei; Li, Wenlian; Su, Zisheng; Peng, Qi Ming; Zhao, Bo; Luo, Yongshi; Jin, Fangming; Yan, Xingwu; Gao, Yuan; Wu, Hairuo; Zhang, Feng; Fan, Di; Wang, Junbo

2014-08-13

We demonstrate highly efficient exciplex delayed-fluorescence organic light-emitting diodes (OLEDs) in which 4,4',4″-tris[3-methylphenyl(phenyl)aminotriphenylamine (m-MTDATA) and 4,7-diphenyl-1,10-phenanthroline (Bphen) were selected as donor and acceptor components, respectively. Our m-MTDATA:Bphen exciplex electroluminescence (EL) mechanism is based on reverse intersystem crossing (RISC) from the triplet to singlet excited states. As a result, an external quantum efficiency (EQE) of 7.79% at 10 mA/cm(2) was observed, which increases by 3.2 and 1.5 times over that reported in Nat. Photonics 2012, 6, 253 and Appl. Phys. Lett. 2012, 101, 023306, respectively. The high EQE would be attributed to a very easy RISC process because the energy difference between the singlet and triplet excited states is almost around zero. The verdict was proven by photoluminescence (PL) rate analysis at different temperatures and time-resolved spectral analysis. Besides, the study of the transient PL process indicates that the presence of an unbalanced charge in exciplex EL devices is responsible for the low EQE and high-efficiency roll-off. When the exciplex devices were placed in a 100 mT magnetic field, the permanently positive magnetoelectroluminescence and magnetoconductivity were observed. The magnetic properties confirm that the efficient exciplex EL only originates from delayed fluorescence via RISC processes but is not related to the triplet-triplet annihilation process.

A trident dithienylethene-perylenemonoimide dyad with super fluorescence switching speed and ratio

NASA Astrophysics Data System (ADS)

Li, Chong; Yan, Hui; Zhao, Ling-Xi; Zhang, Guo-Feng; Hu, Zhe; Huang, Zhen-Li; Zhu, Ming-Qiang

2014-12-01

Photoswitchable fluorescent diarylethenes are promising in molecular optical memory and photonic devices. However, the performance of current diarylethenes is far from satisfactory because of the scarcity of high-speed switching capability and large fluorescence on-off ratio. Here we report a trident perylenemonoimide dyad modified by triple dithienylethenes whose photochromic fluorescence quenching ratio at the photostationary state exceeds 10,000 and the fluorescence quenching efficiency is close to 100% within seconds of ultraviolet irradiation. The highly sensitive fluorescence on/off switching of the trident dyad enables recyclable fluorescence patterning and all-optical transistors. The prototype optical device based on the trident dyad enables the optical switching of incident light and conversion from incident light wavelength to transmitted light wavelength, which is all-optically controlled, reversible and wavelength-convertible. In addition, the trident dyad-staining block copolymer vesicles are observed via optical nanoimaging with a sub-100 nm resolution, portending a potential prospect of the dithienylethene dyad in super-resolution imaging.

A trident dithienylethene-perylenemonoimide dyad with super fluorescence switching speed and ratio.

PubMed

Li, Chong; Yan, Hui; Zhao, Ling-Xi; Zhang, Guo-Feng; Hu, Zhe; Huang, Zhen-Li; Zhu, Ming-Qiang

2014-12-12

Photoswitchable fluorescent diarylethenes are promising in molecular optical memory and photonic devices. However, the performance of current diarylethenes is far from satisfactory because of the scarcity of high-speed switching capability and large fluorescence on-off ratio. Here we report a trident perylenemonoimide dyad modified by triple dithienylethenes whose photochromic fluorescence quenching ratio at the photostationary state exceeds 10,000 and the fluorescence quenching efficiency is close to 100% within seconds of ultraviolet irradiation. The highly sensitive fluorescence on/off switching of the trident dyad enables recyclable fluorescence patterning and all-optical transistors. The prototype optical device based on the trident dyad enables the optical switching of incident light and conversion from incident light wavelength to transmitted light wavelength, which is all-optically controlled, reversible and wavelength-convertible. In addition, the trident dyad-staining block copolymer vesicles are observed via optical nanoimaging with a sub-100 nm resolution, portending a potential prospect of the dithienylethene dyad in super-resolution imaging.

Fluorescence in situ hybridization for phytoplasma and endophytic bacteria localization in plant tissues.

PubMed

Bulgari, Daniela; Casati, Paola; Faoro, Franco

2011-11-01

In the present study, we developed a rapid and efficient fluorescence in situ hybridization assay (FISH) in non-embedded tissues of the model plant Catharanthus roseus for co-localizing phytoplasmas and endophytic bacteria, opening new perspectives for studying the interaction between these microorganisms. Copyright © 2011 Elsevier B.V. All rights reserved.

Design and Development of an Electrostatic Screen Battery for Emission Control (ESBEC)

PubMed Central

Han, Taewon; Mainelis, Gediminas

2017-01-01

Current diesel particulate filters (DPFs) can effectively capture the exhaust particles, but they add to engine backpressure and accumulate particles during their operation, which results in the need to regenerate the DPFs by burning off the collected particles periodically. This regeneration results in aerosol emissions, especially in the 10–30 nanometer size range and contributes to ultrafine particle pollution. In this research, we designed and developed a prototype of a novel diesel exhaust control device: the Electrostatic Screen Battery for Emissions Control (ESBEC). The device features high particle collection efficiency without adding to the exhaust backpressure and without the need for thermal regeneration of the collected particles. The ESBEC consists of a series of metal mesh screens coated with a superhydrophobic substance and an integrated carbon fiber ionizer to charge the incoming particles. Multiple pairs of screens (e.g., 5 pairs) are arranged in a battery, in which one screen of each pair is supplied with high voltage, and the other is grounded, producing electrostatic field produced across the screens. The application of a superhydrophobic coating onto the screens allows easy removal of the collected particles using liquid without the need for thermal regeneration. The current prototypes of the device were tested with fluorescent polystyrene latex (PSL) particles of 0.2 and 1.2 μm in size and at 25 and 105 L/min sampling flow rates. The average collection efficiency was ~87% for 0.2 μm and ~95% for 1.2 μm PSL particles. In addition, the ESBEC was tested with actual diesel exhaust particles; here its performance was verified by visually inspecting deposition of particles on an after-filter with the device ON and OFF. In the next stages of this work, the ESBEC will be challenged with diesel exhaust at different mass concentrations and for different collection time periods. PMID:28983124

Design and Development of an Electrostatic Screen Battery for Emission Control (ESBEC).

PubMed

Han, Taewon; Mainelis, Gediminas

2017-05-01

Current diesel particulate filters (DPFs) can effectively capture the exhaust particles, but they add to engine backpressure and accumulate particles during their operation, which results in the need to regenerate the DPFs by burning off the collected particles periodically. This regeneration results in aerosol emissions, especially in the 10-30 nanometer size range and contributes to ultrafine particle pollution. In this research, we designed and developed a prototype of a novel diesel exhaust control device: the Electrostatic Screen Battery for Emissions Control (ESBEC). The device features high particle collection efficiency without adding to the exhaust backpressure and without the need for thermal regeneration of the collected particles. The ESBEC consists of a series of metal mesh screens coated with a superhydrophobic substance and an integrated carbon fiber ionizer to charge the incoming particles. Multiple pairs of screens (e.g., 5 pairs) are arranged in a battery, in which one screen of each pair is supplied with high voltage, and the other is grounded, producing electrostatic field produced across the screens. The application of a superhydrophobic coating onto the screens allows easy removal of the collected particles using liquid without the need for thermal regeneration. The current prototypes of the device were tested with fluorescent polystyrene latex (PSL) particles of 0.2 and 1.2 μm in size and at 25 and 105 L/min sampling flow rates. The average collection efficiency was ~87% for 0.2 μm and ~95% for 1.2 μm PSL particles. In addition, the ESBEC was tested with actual diesel exhaust particles; here its performance was verified by visually inspecting deposition of particles on an after-filter with the device ON and OFF. In the next stages of this work, the ESBEC will be challenged with diesel exhaust at different mass concentrations and for different collection time periods.

Method of oocyte activation affects cloning efficiency in pigs.

PubMed

Whitworth, Kristin M; Li, Rongfeng; Spate, Lee D; Wax, David M; Rieke, August; Whyte, Jeffrey J; Manandhar, Gaurishankar; Sutovsky, Miriam; Green, Jonathan A; Sutovsky, Peter; Prather, Randall S

2009-05-01

The following experiments compared the efficiency of three fusion/activation protocols following somatic cell nuclear transfer (SCNT) with porcine somatic cells transfected with enhanced green fluorescent protein driven by the chicken beta-actin/rabbit beta-globin hybrid promoter (pCAGG-EGFP). The three protocols included electrical fusion/activation (NT1), electrical fusion/activation followed by treatment with a reversible proteasomal inhibitor MG132 (NT2) and electrical fusion in low Ca(2+) followed by chemical activation with thimerosal/dithiothreitol (NT3). Data were collected at Days 6, 12, 14, 30, and 114 of gestation. Fusion rates, blastocyst-stage mean cell numbers, recovery rates, and pregnancy rates were calculated and compared between protocols. Fusion rates were significantly higher for NT1 and NT2 compared to NT3 (P < 0.05). There was no significant difference in mean nuclear number. Pregnancy rate for NT2 was 100% (n = 19) at all stages collected and was significantly higher than NT1 (71.4%, n = 28; P < 0.05), but was not significantly higher than NT3 (82.6%, n = 23; P < 0.15). Recovery rates were calculated based on the number of embryos, conceptuses, fetuses, or piglets present at the time of collection, divided by the number of embryos transferred to the recipient gilts. Recovery rates between the three groups were not significantly different at any of the stages collected (P > 0.05). All fusion/activation treatments produced live, pCAGG-EGFP positive piglets from SCNT. Treatment with MG132 after fusion/activation of reconstructed porcine embryos was the most effective method when comparing the overall pregnancy rates. The beneficial effect of NT2 protocol may be due to the stimulation of proteasomes that infiltrate donor cell nucleus shortly after nuclear transfer. (c) 2008 Wiley-Liss, Inc.

Measurement of the fluorescence of crop residues: A tool for controlling soil erosion

NASA Technical Reports Server (NTRS)

Daughtry, C. S. T.; Mcmurtrey, J. E., III; Chappelle, E. W.; Hunter, W. J.

1994-01-01

Management of crop residues, the portion of a crop left in the field after harvest, is an important conservation practice for minimizing soil erosion and for improving water quality. Quantification of crop residue cover is required to evaluate the effectiveness of conservation tillage practices. Methods are needed to quantify residue cover that are rapid, accurate, and objective. The fluorescence of crop residue was found to be a broadband phenomenon with emission maxima at 420 to 495 nm for excitations of 350 to 420 nm. Soils had low intensity broadband emissions over the 400 to 690 nm region for excitations of 300 to 600 nm. The range of relative fluorescence intensities for the crop residues was much greater than the fluorescence observed of the soils. As the crop residues decompose their blue fluorescence values approach the fluorescence of the soil. Fluorescence techniques are concluded to be less ambiguous and better suited for discriminating crop residues and soils than reflectance methods. If properly implemented, fluorescence techniques can be used to quantify, not only crop residue cover, but also photosynthetic efficiency in the field.

In vivo tumor-targeted dual-modal fluorescence/CT imaging using a nanoprobe co-loaded with an aggregation-induced emission dye and gold nanoparticles.

PubMed

Zhang, Jimei; Li, Chan; Zhang, Xu; Huo, Shuaidong; Jin, Shubin; An, Fei-Fei; Wang, Xiaodan; Xue, Xiangdong; Okeke, C I; Duan, Guiyun; Guo, Fengguang; Zhang, Xiaohong; Hao, Jifu; Wang, Paul C; Zhang, Jinchao; Liang, Xing-Jie

2015-02-01

As an intensely studied computed tomography (CT) contrast agent, gold nanoparticle has been suggested to be combined with fluorescence imaging modality to offset the low sensitivity of CT. However, the strong quenching of gold nanoparticle on fluorescent dyes requires complicated design and shielding to overcome. Herein, we report a unique nanoprobe (M-NPAPF-Au) co-loading an aggregation-induced emission (AIE) red dye and gold nanoparticles into DSPE-PEG(2000) micelles for dual-modal fluorescence/CT imaging. The nanoprobe was prepared based on a facile method of "one-pot ultrasonic emulsification". Surprisingly, in the micelles system, fluorescence dye (NPAPF) efficiently overcame the strong fluorescence quenching of shielding-free gold nanoparticles and retained the crucial AIE feature. In vivo studies demonstrated the nanoprobe had superior tumor-targeting ability, excellent fluorescence and CT imaging effects. The totality of present studies clearly indicates the significant potential application of M-NPAPF-Au as a dual-modal non-invasive fluorescence/X-ray CT nanoprobe for in vivo tumor-targeted imaging and diagnosis. Copyright © 2014 Elsevier Ltd. All rights reserved.