The problem of atmospheric emission from OH molecules is a long standing problem for near-infrared astronomy. We are now close to solving this problem for the first time with the PRAXIS instrument. PRAXIS is a unique spectrograph which is fed by fibres that remove the OH background, and is optimised specifically to benefit from OH-Suppression. The OH suppression is achieved with fibre Bragg gratings, which were tested successfully on the GNOSIS instrument. The OH lines are suppressed by a factor of ~1000, leading to a reduction of the integrated background of a factor ≈9. A future upgrade to multicore fibre Bragg gratings will further increase this reduction. PRAXIS has had two commissioning runs, with a third commissioning run planned for July 2019, which will be presented at the conference. PRAXIS has a measured throughput of ≈20 %, demonstrating high efficiency in an OH suppression instrument for the first time. Science verification observations of Seyfert galaxies demonstrate the potential of OH suppression.
The problem of atmospheric emission from OH molecules is a long standing problem for near-infrared astronomy. PRAXIS is a unique spectrograph which is fed by fibres that remove the OH background and is optimised specifically to benefit from OH-Suppression. The OH suppression is achieved with fibre Bragg gratings, which were tested successfully on the GNOSIS instrument. PRAXIS uses the same fibre Bragg gratings as GNOSIS in its first implementation, and will exploit new, cheaper and more efficient, multicore fibre Bragg gratings in the second implementation. The OH lines are suppressed by a factor of ∼ 1000, and the expected increase in the
signal-to-noise in the interline regions compared to GNOSIS is a factor of ∼ 9 with the GNOSIS gratings and a
factor of ∼ 17 with the new gratings.
PRAXIS will enable the full exploitation of OH suppression for the first time, which was not achieved by GNOSIS (a retrofit to an existing instrument that was not OH-Suppression optimised) due to high thermal emission, low spectrograph transmission and detector noise. PRAXIS has extremely low thermal emission, through the cooling of all significantly emitting parts, including the fore-optics, the fibre Bragg gratings, a long length of fibre, and the fibre slit, and an optical design that minimises leaks of thermal emission from outside the spectrograph. PRAXIS has low detector noise through the use of a Hawaii-2RG detector, and a high throughput through a efficient VPH based spectrograph. PRAXIS will determine the absolute level of the interline continuum and enable observations of individual objects via an IFU. In this paper we give a status update and report on acceptance tests.
Atmospheric emission from OH molecules is a long standing problem for near-infrared astronomy. PRAXIS is a unique spectrograph, currently in the build-phase, which is fed by a fibre array that removes the OH background. The OH suppression is achieved with fibre Bragg gratings, which were tested successfully on the GNOSIS instrument. PRAXIS will use the same fibre Bragg gratings as GNOSIS in the first implementation, and new, less expensive and more efficient, multicore fibre Bragg gratings in the second implementation. The OH lines are suppressed by a factor of ~1000, and the expected increase in the signal-to-noise in the interline regions compared to GNOSIS is a factor of ~ 9 with the GNOSIS gratings and a factor of ~ 17 with the new gratings. PRAXIS will enable the full exploitation of OH suppression for the first time, which was not achieved by GNOSIS due to high thermal emission, low spectrograph transmission, and detector noise. PRAXIS will have extremely low thermal emission, through the cooling of all significantly emitting parts, including the fore-optics, the fibre Bragg gratings, a long length of fibre, and a fibre slit, and an optical design that minimises leaks of thermal emission from outside the spectrograph. PRAXIS will achieve low detector noise through the use of a Hawaii-2RG detector, and a high throughput through an efficient VPH based spectrograph. The scientific aims of the instrument are to determine the absolute level of the interline continuum and to enable observations of individual objects via an IFU. PRAXIS will first be installed on the AAT, then later on an 8m class telescope.
For the past forty years, optical fibres have found widespread use in ground-based and space-based instruments. In most applications, these fibres are used in conjunction with conventional optics to transport light. But photonics offers a huge range of optical manipulations beyond light transport that were rarely exploited before 2001. The fundamental obstacle to the broader use of photonics is the difficulty of achieving photonic action in a multimode fibre. The first step towards a general solution was the invention of the photonic lantern1 in 2004 and the delivery of high-efficiency devices (< 1 dB loss) five years on2. Multicore fibres (MCF), used in conjunction with lanterns, are now enabling an even bigger leap towards multimode photonics. Until recently, the single-moded cores in MCFs were not sufficiently uniform to achieve telecom (SMF-28) performance. Now that high-quality MCFs have been realized, we turn our attention to printing complex functions (e.g. Bragg gratings for OH suppression) into their N cores. Our first work in this direction used a Mach-Zehnder interferometer (near-field phase mask) but this approach was only adequate for N=7 MCFs as measured by the grating uniformity3. We have now built a Sagnac interferometer that gives a three-fold increase in the depth of field sufficient to print across N ≥ 127 cores. We achieved first light this year with our 500mW Sabre FRED laser. These are sophisticated and complex interferometers. We report on our progress to date and summarize our first-year goals which include multimode OH suppression fibres for the Anglo-Australian Telescope/PRAXIS instrument and the Discovery Channel Telescope/MOHSIS instrument under development at the University of Maryland.
Nanobioconjugates have been synthesized using cadmium selenide quantum dots (QDs), europium complexes (EuCs), and biotin. In those conjugates, long-lived photoluminescence (PL) is provided by the europium complexes, which efficiently transfer energy via Förster resonance energy transfer (FRET) to the QDs in close spatial proximity. As a result, the conjugates have a PL emission spectrum characteristic for QDs combined with the long PL decay time characteristic for EuCs. The nanobioconjugates synthesis strategy and photophysical properties are described as well as their performance in a time-resolved streptavidin-biotin PL assay. In order to prepare the QD-EuC-biotin conjugates, first an amphiphilic polymer has been functionalized with the EuC and biotin. Then, the polymer has been brought onto the surface of the QDs (either QD655 or QD705) to provide functionality and to make the QDs water dispersible. Due to a short distance between EuC and QD, an efficient FRET can be observed. Additionally, the QD-EuC-biotin conjugates’ functionality has been demonstrated in a PL assay yielding good signal discrimination, both from autofluorescence and directly excited QDs. These newly designed QD-EuC-biotin conjugates expand the class of highly sensitive tools for bioanalytical optical detection methods for diagnostic and imaging applications.
GNOSIS has provided the first on-telescope demonstration of a concept to utilize complex aperioidc fiber Bragg
gratings to suppress the 103 brightest atmospheric hydroxyl emission doublets between 1.47-1.7 μm. The unit is
designed to be used at the 3.9-meter Anglo-Australian Telescope (AAT) feeding the IRIS2 spectrograph. Unlike
previous atmospheric suppression techniques GNOSIS suppresses the lines before dispersion. We present the
results of laboratory and on-sky tests from instrument commissioning. These tests reveal excellent suppression
performance by the gratings and high inter-notch throughput, which combine to produce high fidelity OH-free
spectra.
Semiconductor quantum dots (QDs) are highly interesting fluorophores for a large variety of spectroscopic applications.
Although their fluorescence properties are well investigated, accurate size determination of QDs is still a problem. TEM
techniques can image the inorganic core/shell system of QDs, but size determination of polymer coated QDs is difficult.
SEC (size exclusion chromatography) compares the QD size only with standard polymers and their sizes, and is therefore
not easy to use on nanoparticles. As QDs are fluorescent, single molecule spectroscopy methods such as fluorescence
correlation spectroscopy (FCS) can be used to determine QDs diffusion coefficients and hence their hydrodynamic radii.
Moreover, this method for size determination requires only very low QD concentrations, which is a mayor advantage
compared to other techniques such as dynamic light scattering.
Within our contribution we present the size determination of commercially available and self-modified QDs with FCS.
The commercial QDs (QD525, QD565, QD605, QD655 and QD705 - purchased from Invitrogen Inc.) have a rather
thick polymer shell and are functionalized with streptavidin, biotin or carboxylic groups. The self-modified QDs consist
of the same commercial core/shell QDs and are modified with a polymer shell and several bio-functionalization groups.
For all nanoparticles the diffusion coefficients were measured by FCS and the hydrodynamic radii were calculated according
to the Stokes-Einstein equation. The obtained results are in good agreement with the size information provided
by Invitrogen Inc., which demonstrates that FCS is an important technique for QD size determination at very low concentrations.
Semiconductor nanocrystals (quantum dots - QDs) possess unique photophysical properties that make them highly interesting
for many biochemical applications. Besides their common use as fluorophores in conventional spectroscopy and
microscopy, QDs are well-suited for studying Förster resonance energy transfer (FRET). Size-dependent broadband
absorption and narrow emission bands offer several advantages for the use of QDs both as FRET donors and acceptors.
QD-based FRET pairs can be efficiently used as biological and chemical sensors for highly sensitive multiplexed detection.
In this contribution we present the use of several commercially available QDs (Qdot® Nanocrystals - Invitrogen) as
FRET donors in combination with commercial organic dyes as FRET acceptors. In order to investigate the FRET process
within our donor-acceptor pairs, we used biotinylated QDs and streptavidin-labeled dyes. The well-known biotinstreptavidin
molecular recognition enables effective FRET from QDs to dye molecules and provides defined distances
between donor and acceptor. Steady-state and time-resolved fluorescence measurements were performed in order to
investigate QD-to-dye FRET. Despite a thick polymer shell around the QDs, our results demonstrate the potential of
these QDs as efficient donors both for steady-state and time-resolved FRET applications in nano-biotechnology.
GNOSIS is an OH suppression unit to be used in conjunction with existing spectrographs. The OH suppression
is achieved using fibre Bragg gratings (FBGs), and will deliver the darkest near-infrared background of any
ground-based instrument. Laboratory and on-sky tests demonstrate that FBGs can suppress OH lines by 30dB
whilst maintaing > 90% throughput between the lines, resulting in a 4 mag decrease in the background.
In the first implementation GNOSIS will feed IRIS2 on the AAT. It will consist of a seven element lenslet
array, covering 1.4" on the sky, and will suppress the 103 brightest OH lines between 1.47 and 1.70 μm. Future
upgrades will include J-band suppression and implementation on an 8m telescope.
Supercontinuum white light sources (SCLS) are intense, spatially coherent laser sources with a very broad and flat
spectral energy distribution which have very quickly found ubiquitous use in optical laboratories. As photonics is now
providing more and more applications for astronomical instrumentation, the possible use of SCLS as a calibration light
source for spectroscopy has been tested. A standard industrial SCLS was coupled to the calibration unit of the PMAS
integral field spectrophotometer and compared directly to the PMAS standard tungsten filament lamp that is normally
used for calibration exposures. We report on comparative measurements concerning flux, spectral energy distribution,
and temporal stability.
Spatially resolved sensing of molecular oxygen is important for many biological and environmental applications. For this
purpose, time-resolved fluorescence measurements were combined with optical time domain reflectometry (OTDR), a
technique which was primarily developed for inspections of optical fiber lines. To achieve spatial resolution of some
meters, which are typical for commercial OTDR instruments, the lifetimes of the sensor dyes must be within the range of
some nanoseconds, which is much shorter than the decay times of common phosphorescent oxygen probes. Therefore,
the measurements were performed with a novel fluorescent triangular-[4]phenylene sensor dye. The fluorescence decay
times are around 80 ns in absence of oxygen and around 20 ns in the presence of air. The [4]phenylene sensor dye was
applied in toluene solution as well as immobilized in a polymer film. Using a branched model fiber line, oxygen
measurements were carried out in a micro- to millimolar concentration range. Oxygen-dependent fluorescence decay
times measured with OTDR in toluene were verified by use of time-correlated single photon counting (TCSPC). The
Stern-Volmer plots obtained for fluorescence quenching of sensor dyes dissolved in toluene solution and polymer-based
sensor spots show good linearity.
An optical multiplexed homogeneous (liquid phase) immunoassay based on FRET from a terbium complex to eight
different fluorescent dyes is presented. We achieved highly sensitive parallel detection of four different lung cancer
specific tumor markers (CEA, NSE, SCC and CYFRA21-1) within a single assay and show a proof-of-principle for 5-
fold multiplexing. The method is well suited for fast and low-cost miniaturized point-of-care testing as well as for highthroughput
screening in a broad range of in-vitro diagnostic applications.
We present the use of luminescent terbium complexes (LTCs) as FRET donors and luminescent semiconductor quantum
dots (QDs) as FRET acceptors for spectroscopic ruler measurements. The LTCs were labeled to polyhistidine-appended
peptides which self-assembled onto three different QDs allowing optical multiplexed measurements. Forster distances
were in the range of 60-75 Å and FRET efficiencies of up to 97 % were realized. Time-resolved analysis allowed the
determination of donor-acceptor separation distances. The results suggests the efficient use of our LTC-to-QD FRET
systems for multiplexed optical size determination, molecular ruler measurements and multi-parameter diagnostics.
In this contribution we present the application of five different commercially available semiconductor core/shell quantum
dots (Qdot® Nanocrystals - Invitrogen Corp.) as multiplexing FRET acceptors together with a commercial
supramolecular terbium complex (Lumi4®-Tb - Lumiphore Inc.) as donor in a homogeneous immunoassay format. To
realize the molecular recognition necessary for a FRET assay, the terbium complex was labeled to streptavidin (sAv-
Lumi4-Tb) and the QDs were surface functionalized with biotin (Biot-QD). The biotin-streptavidin binding serves as a
proof-of-principle representative for many biological interactions taking place on the nanometer scale (e.g.
immunoassays). The presented FRET system can be efficiently used for the detection of inter- and intramolecular
processes for clinical diagnostics and biomedical spectroscopy as well as molecular ruler applications and microscopy.
Homogeneous immunoassays have the benefit that they do not require any time-consuming separation steps. FRET is
one of the most sensitive homogeneous methods used for immunoassays. Due to their extremely strong absorption over a
broad wavelength range the use of quantum dots as FRET acceptors allows for large Foerster radii, an important
advantage for assays in the 5 to 10 nm distance range. Moreover, because of their size-tunable emission, quantum dots of
different sizes can be used with a single donor for the detection of different analytes (multiplexing). As the use of
organic dyes with short fluorescence decay times as donors is known to be inefficient with quantum dot acceptors,
lanthanide complexes with long luminescence decays are very efficient alternatives.
In this contribution we present the application of commercially available biocompatible CdSe/ZnS core/shell quantum
dots as multiplexing FRET acceptors together with a single terbium complex as donor in a homogeneous immunoassay
system. Foerster radii of 10 nm and FRET efficiencies of 75 % are demonstrated. The high sensitivity of the terbium-toquantum
dot FRET assay is shown by sub-100-femtomolar detection limits for two different quantum dots (emitting at
605 and 655 nm) within the same biotin-streptavidin assay. Direct comparison to the FRET immunoassay "gold standard" (FRET from Eu-TBP to APC) yields a three orders of magnitude sensitivity improvement, demonstrating the
big advantages of quantum dots not only for multiplexing but also for highly sensitive nanoscale analysis.
In this study two issues are addressed, namely laser ionisation of selected nitroaromatic compounds (NAC) and the characterisation of their anions by photodetachment (PD) spectroscopy. Laser ionisation of the NAC at λ = 226.75 nm is investigated by ion mobility (IM) spectrometry at atmospheric pressure. The main product after laser ionisation is the reactive NO+ ion formed in a sequence of photofragmentation and multiphoton ionisation processes. NO+ is trapped by specific ion molecule reactions (IMR). Alternatively, NO, added as laser dopant, can directly be ionised. The formed NO+ reacts with the NAC under complex formation. This allows fragmentless NAC detection. The combination of IM spectrometry and PD spectroscopy provides real-time characterisation of the anions in the IM spectrum. This is useful to differentiate between NAC and interfering substances and, thus, to reduce false-positive detections of NAC. The electrons detached by the PD laser at λ = 532 nm are detected in the same spectrum as the anions. The potential of PD-IM spectrometry in terms of cross section determination, analytical improvements, tomographic mapping, spatial hole burning etc., is outlined.
innoFSPEC Potsdam is presently being established as in interdisciplinary innovation center for fiber-optical
spectroscopy and sensing, hosted by Astrophysikalisches Institut Potsdam and the Physical Chemistry group of Potsdam
University, Germany. The center focuses on fundamental research in the two fields of fiber-coupled multi-channel
spectroscopy and optical fiber-based sensing. Thanks to its interdisciplinary approach, the complementary methodologies
of astrophysics on the one hand, and physical chemistry on the other hand, are expected to spawn synergies that
otherwise would not normally become available in more standard research programmes. innoFSPEC targets future
innovations for next generation astrophysical instrumentation, environmental analysis, manufacturing control and
process monitoring, medical diagnostics, non-invasive imaging spectroscopy, biopsy, genomics/proteomics, high-throughput
screening, and related applications.
Phototropic microalgae have a large potential for producing valuable substances for the feed, food, cosmetics, pigment, bioremediation, and
pharmacy industries as well as for biotechnological processes. Today it is estimated that the microalgal aquaculture worldwide production
is 5000 tons of dry matter per year (not taking into account processed products) making it an approximately $1.25 billion U.S. per year
industry. For effective observation of the photosynthetic growth processes, fast on-line sensor systems that analyze the
relevant biological and technical process parameters are preferred. The optical properties of the microalgae culture influence the
transport of light in the photobioreactor and can be used to extract relevant information for efficient cultivation practices. Microalgae
cultivation media show a combination of light absorption and scattering, which are influenced by the concentrations and the physical and
chemical properties of the different absorbing and scattering species (e.g. pigments, cell components, etc.). Investigations with frequency
domain photon density waves (PDW) allow for the examination of absorption and scattering properties of turbid media, namely the absorption
and reduced scattering coefficient. The reduced scattering coefficient can be used to characterize physical and morphological properties of
the medium, including the cell concentration, whereas the absorption coefficient correlates with the pigment content.
Nannochloropsis oculata, a single-cell species of microalgae, were examined in a nutrient solution with photon density waves. The
absorption and reduced scattering coefficients were experimentally determined throughout the cultivation process, and applied to gain
information about the cell concentration and average cell radius.
Optical measurements of molecular oxygen are based on phosphorescent dyes with decay times dependent on the
ambient oxygen content. Additionally, the phosphorescence decay times are affected by the temperature. In this work,
we present miniaturized fiber optical probes, which are able to determine both ambient oxygen content and temperature
simultaneously.
Homogeneous FRET (Forster Resonance Energy Transfer) fluoroimmunoassays (FIA) allow fast, inexpensive
and highly sensitive monitoring of biochemical processes occurring on the nanometer scale. The technique
is widely applied in high throughput screening (HTS) for in vitro diagnostics (IVD). Quantum dots (qdots) are usually applied as energy donors for FRET experiments in solution. In this contribution we show that commercially available biotinylated CdSe/ZnS core/shell qdots (Qdot 665 Biotin conjugate, Invitrogen Corp., USA) are excellent FRET acceptors in a time-resolved FIA based on interaction with lanthanide complex-labeled streptavidin as energy donor. The energy transfer experiments were performed on a modified commercial FIA reader system (KRYPTOR, Cezanne SA, France) using three di.erent lanthanide chelates (1 of terbium and 2 of europium). All three FRET donors showed efficient energy transfer to the qdots, evidenced both by nanocrystals emission sensitization and by a thousand fold increase of the qdot luminescence decay time, reaching some
hundreds of microseconds. In a control experiment the unlabeled donors were used to rule out dynamic energy
transfer between lanthanides and qdots. Due to the very high qdot absorption extremely large Forster radii of
104 A for terbium and 96 A for europium were achieved. FRET efficiency was up to 67 % and sub-picomolar detection limits were obtained for qdots in this type of homogeneous FIA. The use of qdots as energy acceptors potentially offers a broad scope of scientific and commercial applications such as ultra-sensitive FIA, the study of interactions within very large molecules, biomedical HTS and multiplexed analysis.
We demonstrate the improvement of fluorescence immunoassay (FIA) diagnostics in deploying a newly developed compact diode-pumped solid state (DPSS) laser with emission at 315 nm. The laser is based on the quasi-three-level transition in Nd:YAG at 946 nm. The pulsed operation is either realized by an active Q-switch using an electro-optical device or by introduction of a Cr4+:YAG saturable absorber as passive Q-switch element. By extra-cavity second harmonic generation in different nonlinear crystal media we obtained blue light at 473 nm. Subsequent mixing of the fundamental and the second harmonic in a β-barium-borate crystal provided pulsed emission at 315 nm with up to 20 μJ maximum pulse energy and 17 ns pulse duration. Substitution of a nitrogen laser in a FIA diagnostics system by the DPSS laser succeeded in considerable improvement of the detection limit. Despite significantly lower pulse energies (7 μJ DPSS laser versus 150 μJ nitrogen laser), in preliminary investigations the limit of detection was reduced by a factor of three for a typical FIA.
The detection of polar molecules, like ketones and ethers, in a laser ion mobility spectrometer was investigated. Because
the direct multiphoton ionization (MPI) for these compounds shows too high limits of detection (LOD) and intensive
fragmentation of the molecular ions, alternative ionization methods based on ion-molecule-reactions (IMR) were
investigated. These ionization methods should retain the advantages of the laser ionization. As examples for IMR two
reaction classes, proton transfer reactions (PTR) and complex formation reactions (CFR), were studied. The PTR are
based in a first step on the proton transfer from toluene radical cations to polar molecules. In a second step protonated
dimers are formed. The CFR are characterized by the complex formation between aniline or phenol radical cations and
polar molecules. All products are formed at atmospheric pressure and are characterized by transfer into a time-of-flight
mass spectrometer. In both IMR the ionic reactants are formed selectively by 1+1 REMPI. The rates of the following
IMR are near to the collision limit. Therefore the reactions are very efficient. The LOD for the analysis of selected
ketones and ethers by IMR are in the low ppb-range, much lower than the corresponding LOD for direct MPI. The
required laser intensities for the IMR are up to a factor of 1000 lower than the laser intensities for direct MPI. The
fragmentation of the product ions is much lower for both IMR in comparison to direct MPI. The IMR allow the
quantitative analysis of substance mixtures.
Near-infrared (NIR) absorption spectroscopy with tunable diode lasers allows the simultaneous detection of the three
most important isotopologues of carbon dioxide (12CO2, 13CO2, 12C18O16O) and carbon monoxide (12CO,13CO, 12C18O).
The flexible and compact fiber-optic tunable diode laser absorption spectrometer (TDLAS) allows selective
measurements of CO2 and CO with high isotopic resolution without sample preparation since there is no interference
with water vapour. For each species, linear calibration plots with a dynamic range of four orders of magnitude and
detection limits (LOD) in the range of a few ppm were obtained utilizing wavelength modulation spectroscopy (WMS)
with balanced detection in a Herriott-type multipass cell. The high performance of the apparatus is illustrated by fill-evacuation-
refill cycles.
KEYWORDS: Fluorescence resonance energy transfer, Luminescence, Quantum dots, Terbium, Lanthanides, Quantum efficiency, Energy transfer, Ions, Absorption, Resonance energy transfer
Due to their extraordinary photophysical properties CdSe/ZnS core/shell nanocrystals (quantum dots) are excellent luminescence dyes for fluorescence resonance energy transfer (FRET) systems. By using a supramolecular lanthanide complex with central terbium cation as energy donor, we show that commercially available biocompatible biotinilated quantum dots are excellent energy acceptors in a time-resolved FRET fluoroimmunoassay (FRET-FIA) using streptavidin-biotin binding as biological recognition process. The efficient energy transfer is demonstrated by quantum dot emission sensitization and a thousandfold increase of the nanocrystal luminescence decay time. A Foerster Radius of 90 Å and a picomolar detection limit were achieved in quantum dot borate buffer. Regarding biological applications the influence of bovine serum albumin (BSA) and sodium azide (a frequently used preservative) to the luminescence behaviour of our FRET-system is reported.
KEYWORDS: Fluorescence resonance energy transfer, Luminescence, Europium, Energy transfer, Quantum dots, Molecules, Absorption, Resonance energy transfer, Optical properties, Energy efficiency
Quantum dots (QDs) are common as luminescing markers for imaging in biological applications because their optical properties seem to be inert against their surrounding solvent. This, together with broad and strong absorption bands and intense, sharp tuneable luminescence bands, makes them interesting candidates for methods utilizing Forster Resonance Energy Transfer (FRET), e. g. for sensitive homogeneous fluoroimmunoassays (FIA). In this work we demonstrate energy transfer from Eu3+-trisbipyridin (Eu-TBP) donors to CdSe-ZnS-QD acceptors in solutions with and without serum. The QDs are commercially available CdSe-ZnS core-shell particles emitting at 655 nm (QD655). The FRET system was achieved by the binding of the streptavidin conjugated donors with the biotin conjugated acceptors. After excitation of Eu-TBP and as result of the energy transfer, the luminescence of the QD655 acceptors also showed lengthened decay times like the donors. The energy transfer efficiency, as calculated from the decay times of the bound and the unbound components, amounted to 37%. The Forster-radius, estimated from the absorption and emission bands, was ca. 77Å. The effective binding ratio, which not only depends on the ratio of binding pairs but also on unspecific binding, was obtained from the donor emission dependent on the concentration. As serum promotes unspecific binding, the overall FRET efficiency of the assay was reduced. We conclude that QDs are good substitutes for acceptors in FRET if combined with slow decay donors like Europium. The investigation of the influence of the serum provides guidance towards improving binding properties of QD assays.
Two examples of our biophotonic research utilizing nanoparticles are presented, namely laser-based fluoroimmuno analysis and in-vivo optical oxygen monitoring. Results of the work include significantly enhanced sensitivity of a homogeneous fluorescence immunoassay and markedly improved spatial resolution of oxygen gradients in root nodules of a legume species.
The performance of a home-built tunable diode laser (TDL) spectrometer has been optimized regarding multi-line detection of carbon dioxide in natural gases. In the regime of the (3001)III;←(000) band of 12CO2 around 1.6 μm, the dominating isotope species 12CO2, 13CO2, and12C18O16O were detected simultaneously. In contrast to most established techniques, selective measurements are performed without any sample preparation. This is possible since the CO2 detection is free of interference from water, ubiquitous in natural gases. Detection limits in the range of a few ppmv were obtained for each species utilizing wavelength modulation (WM) spectroscopy with balanced detection in a long-path absorption cell set-up. Linear calibration plots cover a dynamic range of four orders of magnitude, allowing for quantitative CO2 detection in various samples, like soil and breath gas. High isotopic resolution enables the excellent selectivity, sensitivity, and stability of the chosen analytical concept. The obtained isotopic resolution of typically ± 1.0 % and ± 1.5 % (for 3 vol. % and 0.7 vol. % of CO2, respectively) offers a promising analytical tool for isotope-ratio determination of carbon dioxide in soil gas. Preliminary experiments on soil respiration for the first time combine the on-line quantification of the overall carbon dioxide content with an optode sensor and isotopic determination (TDL system) of natural gas species.
The drift time spectra of polycyclic aromatic hydrocarbons (PAH), alkylbenzenes and alkylphenylethers were recorded with a laser-based ion mobility (IM) spectrometer. The ion mobilities of all compounds were determined in helium as drift gas. This allows the calculation of the diffusion cross sections (Ωcalc) on the basis of the exact hard sphere scattering model (EHSSM) and their comparison with the experimentally determined diffusion cross sections (Ωexp). These Ωexp/Ωcalc-correlations are presented for molecules with a rigid structure like PAH and prove the reliability of the theoretical model and experimental method. The increase of the selectivity of IM spectrometry is demonstrated using resonance enhanced multiphoton ionisation (REMPI) at atmospheric pressure, realized by tuneable lasers. The REMPI spectra of nine alkylbenzenes and alkylphenylethers are investigated. On the basis of these spectra, the complete qualitative distinction of eight compounds in a mixture is shown. These experiments are extended to alkylbenzene isomer mixtures.
In this paper results of LIF and DR measurements of diesel- fuel contaminated natural and model soils are presented. In order to characterize the influence of soil optical properties on LIF signal intensities (IF), the reflectances (R) of the soils are taken into account. It is demonstrated that using 'normalized' LIF signal intensities IF/R significantly reduced the influence of the variability of the matrices. For various natural and model soils contaminated with 5000 ppm diesel fuel the ratio of IF/R is shown to be approximately constant. Therefore, the calibration with 'normalized' LIF signal intensities allows the quantification of a contaminant in different soils.
Time-resolved emission spectra of humic substances of different origin have been recorded in order to extend our previous spectroscopic characterization of humic and fulvic acids in aqueous solution with stationary UV/Vis-absorption and fluorescence measurements. Instead of applying a fitting procedure, decay times are introduced as simple and robust experimental parameters for the quantitative description of fluorescence decays. The analysis of time-resolved emission spectra indicates a wavelength dependence of the fluorescence investigations of the interactions between pyrene and various humic substances in water. For all humic substances the quenching of pyrene fluorescence is dominated by a static quenching process, i.e. by a ground-state complexation. A significant influence of pH on the fluorescence quenching efficiencies is found, whereas no long-term changes of the interaction between pyrene and a humic substance are discernible. Time-resolved triplet- triplet absorption measurements reveal a decrease of chrysene triplet lifetime due to the presence of humic substances.
Photophysical properties of the widely used fluorescence tracers eosin, uranin and pyranine were determined in water at different pH. In order to elucidate the potential of LIF spectroscopy for in-situ measurements, calibration curves of uranin in different waters have been measured and the influence of other components in natural waters and of soil on the LIF signals was determined. Furthermore, LIF spectroscopy was applied to the investigation of sorption processes in water/soil-mixtures and of transport processes in soil columns.
Extending our stationary fluorescence and absorption measurements of petroleum products in cyclohexane solution, time-resolved measurements of the fluorescence decay are presented in this paper. Several parameters suitable for the description of fluorescence decay curves of petroleum products are discussed in detail. Measurements of time- resolved emission spectra indicate an emission-wavelength dependence of the decay curves. We have found that the fluorescence emission of petroleum products is significantly influenced by oxygen. Applying a Stern-Volmer analysis it is possible to obtain Stern-Volmer constants Ksv and quenching constants kq for the quenching of the petroleum product fluorescence by molecular oxygen. In addition, the excitation-wavelength dependence of stationary fluorescence as well as the concentration dependence of the effective extinction coefficients have been investigated.
In the present paper results of our LIF measurements of mineral oils on soil surfaces are briefly described. In order to characterize the influence of the analytes on the LIF signals we have measured the photophysical properties of different mineral oils in solution. An attempt to correlate the analytical parameters of the LIF calibration curves on soil surfaces with the photophysical properties in solution is presented.
From the fluorescence analysis of PAC-doped model oils on alumina, quartz sand, and soil surfaces LIF intensity/concentration-relationships (calibration curves) were obtained. It was found that LIF measurements directly from soil surfaces allow the in-situ analysis of PAC contaminations in environmentally relevant concentration ranges. However, the LIF signals measured cannot directly be related to photophysical parameters of the PAC in solution. Also reported are results of preliminary investigations of the long-term behavior and the depth profile of perylene or fluoranthene LIF signals from surfaces reflecting mass transport and sorption processes.
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