TY - JOUR A1 - Napp, J. A1 - Behnke, Thomas A1 - Fischer, L. A1 - Würth, Christian A1 - Wottawa, M. A1 - Katschinski, D.M. A1 - Alves, F. A1 - Resch-Genger, Ute A1 - Schäferling, M. T1 - Targeted luminescent near-infrared polymer-nanoprobes for in vivo imaging of tumor hypoxia N2 - Polystyrene nanoparticles (PS-NPs) were doped with an oxygen-sensitive near-infrared (NIR)-emissive palladium meso-tetraphenylporphyrin and an inert reference dye which are both excitable at 635 nm. The nanosensors were characterized with special emphasis on fundamental parameters such as absolute photoluminescence quantum yield and fluorescence lifetime. The PS-NPs were employed for ratiometric dual-wavelength and lifetime-based photoluminescent oxygen sensing. They were efficiently taken up by cultured murine alveolar macrophages, yielding a characteristic and reversible change in ratiometric response with decreasing oxygen concentration. This correlated with the cellular hypoxic status verified by analysis of hypoxia inducible factor-1α (HIF-1α) accumulation. In addition, the surface of PS-NPs was functionalized with polyethylene glycol (PEG) and the monoclonal antibody herceptin, and their binding to HER2/neu-overexpressing tumor cells was confirmed in vitro. First experiments with tumor-bearing mouse revealed a distinctive ratiometric response within the tumor upon hypoxic condition induced by animal sacrifice. These results demonstrate the potential of these referenced NIR nanosensors for in vitro and in vivo imaging that present a new generation of optical probes for oncology. KW - Fluorescence KW - NIR KW - Nanoparticles KW - Microparticles KW - Imaging KW - Cancer KW - Oxygene PY - 2011 DO - https://doi.org/10.1021/ac201870b SN - 0003-2700 SN - 1520-6882 VL - 83 IS - 23 SP - 9039 EP - 9046 PB - American Chemical Society CY - Washington, DC AN - OPUS4-24987 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Behnke, Thomas A1 - Laux, Eva-Maria A1 - Hoffmann, Katrin A1 - Peters, S. A1 - Haueisen, J. A1 - Klemm, M. A1 - Resch-Genger, Ute T1 - Simple approaches to fluorescence lifetime standards using dye-quencher pairs N2 - Photoluminescence techniques are amongst the most widely used tools in the material and life sciences, with new and exciting applications continuously emerging. Advantages include their comparative ease of use, unique sensitivity, non-invasive character, and potential for multiplexing, remote sensing, and miniaturization. Increasing applications of fluorescence techniques in the life sciences and emerging medical applications of fluorescence microscopic techniques including 1P and 2P fluorescence microscopy combined with fluorescence lifetime imaging (FLIM) in e.g. in vivo eye diagnostics boosted the demand for robust, easy-to-use, and reliable fluorescence standards to ensure the reliability and comparability of fluorescence data. This includes fluorescence standards for the consideration of instrument-specific spectral and intensity distortions of measured signals and instrument performance validation, fluorescence intensity standards for the quantification of measured intensities and for signal referencing, and lately, also fluorescence lifetime standards. T2 - BMT 2012 - 46. DGBMT Jahrestagung CY - Jena, Germany DA - 16.09.2012 PY - 2012 DO - https://doi.org/10.1515/bmt-2012-4499 SN - 0013-5585 SN - 1862-278X VL - 57 IS - Suppl. 1 SP - 613 PB - De Gruyter CY - Berlin AN - OPUS4-27682 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Natte, Kishore A1 - Behnke, Thomas A1 - Orts Gil, Guillermo A1 - Würth, Christian A1 - Friedrich, Jörg Florian A1 - Österle, Werner A1 - Resch-Genger, Ute T1 - Synthesis and characterisation of highly fluorescent core-shell nanoparticles based on Alexa dyes N2 - Current and future developments in the emerging field of nanobiotechnology are closely linked to the rational design of novel fluorescent nanomaterials, e.g. for biosensing and imaging applications. Here, the synthesis of bright near infrared (NIR)-emissive nanoparticles based on the grafting of silica nanoparticles (SNPs) with 3-aminopropyl triethoxysilane (APTES) followed by covalent attachment of Alexa dyes and their subsequent shielding by an additional silica shell are presented. These nanoparticles were investigated by dynamic light scattering (DLS), transmission electron microscopy (TEM) and fluorescence spectroscopy. TEM studies revealed the monodispersity of the initially prepared and fluorophorelabelled silica particles and the subsequent formation of raspberry-like structures after addition of a silica precursor.Measurements of absolute fluorescence quantum yields of these scattering particle suspensions with an integrating sphere setup demonstrated the influence of dye labelling density-dependent fluorophore aggregation on the signaling behaviour of such nanoparticles. KW - Silica KW - Alexa dyes KW - Fluorescent particles KW - Quantum yields KW - Nanoparticles KW - Protective shell KW - Nanobiotechnology PY - 2012 DO - https://doi.org/10.1007/s11051-011-0680-9 SN - 1388-0764 SN - 1572-896X VL - 14 IS - 2 SP - 680-1 - 680-10 PB - Kluwer CY - Dordrecht AN - OPUS4-25872 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Huber, Alexandra A1 - Behnke, Thomas A1 - Würth, Christian A1 - Jäger, Christian A1 - Resch-Genger, Ute T1 - Spectroscopic characterization of coumarin-stained beads: quantification of the number of fluorophores per particle with solid-state 19F-NMR and measurement of absolute fluorescence quantum yields N2 - The rational design of nano- and micrometer-sized particles with tailor-made optical properties for biological, diagnostic, and photonic applications requires tools to characterize the signal-relevant properties of these typically scattering bead suspensions. This includes methods for the preferably nondestructive quantification of the number of fluorophores per particle and the measurement of absolute fluorescence quantum yields and absorption coefficients of suspensions of fluorescent beads for material performance optimization and comparison. Here, as a first proof-of-concept, we present the first time determination of the number of dye molecules per bead using nondestructive quantitative (19F) NMR spectroscopy and 1000 nm-sized carboxylated polystyrene particles loaded with varying concentrations of the laser dye coumarin 153 containing a CF3 group. Additionally, the signal-relevant optical properties of these dye-loaded particles were determined in aqueous suspension in comparison to the free dye in solvents of different polarity with a custom-built integrating sphere setup that enables spectrally resolved measurements of emission, transmission, and reflectance as well absolute fluorescence quantum yields. These measurements present an important step toward absolute brightness values and quantitative fluorescence analysis with particle systems that can be exploited, for example, for optical imaging techniques and different fluorescence assays as well as for the metrological traceability of fluorescence methods. PY - 2012 DO - https://doi.org/10.1021/ac3000682 SN - 0003-2700 SN - 1520-6882 VL - 84 IS - 8 SP - 3654 EP - 3661 PB - American Chemical Society CY - Washington, DC AN - OPUS4-25873 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Mathejczyk, J. A1 - Napp, J. A1 - Hoffmann, Katrin A1 - Würth, Christian A1 - Alves, F. A1 - Schäferling, M. A1 - Resch-Genger, Ute T1 - Bioimaging with rationally designed NIR-flourescent nanosensors T2 - NAMDIATREAM Winter School on Bioimaging, sensing and therapeutic applications of nanomaterials CY - Villars-sur-Ollon, Switzerland DA - 2013-03-03 PY - 2013 AN - OPUS4-27808 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Resch-Genger, Ute A1 - Napp, J. A1 - Alves, F. A1 - Schäferling, M. T1 - NIR-Emissive O2-Nanosensors for Biomarker Imaging T2 - Biosensor Symposium CY - Wildau, Germany DA - 2013-03-10 PY - 2013 AN - OPUS4-27810 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Felbeck, Tom A1 - Behnke, Thomas A1 - Hoffmann, Katrin A1 - Grabolle, Markus A1 - Lezhnina, M.M. A1 - Kynast, U.H. A1 - Resch-Genger, Ute T1 - Nile-red-nanoclay hybrids: Red emissive optical probes for use in aqueous dispersion N2 - Water-dispersible and (bio)functionalizable nanoclays have a considerable potential as inexpensive carriers for organic molecules like drugs and fluorophores. Aiming at simple design strategies for red-emissive optical probes for the life sciences from commercial precursors with minimum synthetic effort, we systematically studied the dye loading behavior and stability of differently functionalized laponites. Here, we present a comprehensive study of the absorption and emission properties of the red emissive hydrophobic and neutral dye Nile Red, a well-known polarity probe, which is almost insoluble and nonemissive in water. Adsorption of this probe onto disk-shaped nanoclays was studied in aqueous dispersion as function of dye concentration, in the absence and presence of the cationic surfactant cetyltrimethylammonium bromide (CTAB) assisting dye loading, and as a function of pH. This laponite loading strategy yields strongly fluorescent nanoclay suspensions with a fluorescence quantum yield of 0.34 at low dye loading concentration. The dye concentration-, CTAB-, and pH-dependent absorption, fluorescence emission, and fluorescence excitation spectra of the Nile-Red6#8211;nanoclay suspensions suggest the formation of several Nile Red species including emissive Nile Red monomers facing a polar environment, nonemissive H-type dimers, and protonated Nile Red molecules that are also nonfluorescent. Formation of all nonemissive Nile Red species could be suppressed by modification of the laponite with CTAB. This underlines the great potential of properly modified and functionalized laponite nanodisks as platform for optical probes with drug delivery capacities, for example, for tumor and therapy imaging. Moreover, comparison of the Nile Red dimer absorption spectra with absorption spectra of previously studied Nile Red aggregates in dendrimer systems and micelles and other literature systems reveals a considerable dependence of the dimer absorption band on microenvironment polarity which has not yet been reported so far for H-type dye aggregates. KW - Nile Red KW - Dye KW - Laponite KW - Nanoclay KW - Photoluminescence KW - Fluorescence KW - Polarity probe KW - Aggregate KW - Dimer PY - 2013 UR - http://pubs.acs.org/doi/pdf/10.1021/la402165q DO - https://doi.org/10.1021/la402165q SN - 0743-7463 SN - 1520-5827 VL - 29 IS - 36 SP - 11489 EP - 11497 PB - American Chemical Society CY - Washington, DC AN - OPUS4-29017 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hoffmann, Katrin A1 - Behnke, Thomas A1 - Drescher, Daniela A1 - Kneipp, Janina A1 - Resch-Genger, Ute T1 - Near-infrared-emitting nanoparticles for lifetime-based multiplexed analysis and imaging of living cells N2 - The increase in information content from bioassays and bioimaging requires robust and efficient strategies for the detection of multiple analytes or targets in a single measurement, thereby addressing current health and security concerns. For fluorescence techniques, an attractive alternative to commonly performed spectral or color multiplexing presents lifetime multiplexing and the discrimination between different fluorophores based on their fluorescence decay kinetics. This strategy relies on fluorescent labels with sufficiently different lifetimes that are excitable at the same wavelength and detectable within the same spectral window. Here, we report on lifetime multiplexing and discrimination with a set of nanometer-sized particles loaded with near-infrared emissive organic fluorophores chosen to display very similar absorption and emission spectra, yet different fluorescence decay kinetics in suspension. Furthermore, as a first proof-of-concept, we describe bioimaging studies with 3T3 fibroblasts and J774 macrophages, incubated with mixtures of these reporters employing fluorescence lifetime imaging microscopy. These proof-of-concept measurements underline the potential of fluorescent nanoparticle reporters in fluorescence lifetime multiplexing, barcoding, and imaging for cellular studies, cell-based assays, and molecular imaging. KW - Fluorescence lifetime imaging microscopy KW - FLIM KW - Lifetime multiplexing KW - Near infrared KW - NIR KW - Cell imaging KW - Nanoparticles PY - 2013 DO - https://doi.org/10.1021/nn4029458 SN - 1936-0851 VL - 7 IS - 8 SP - 6674 EP - 6684 PB - ACS Publ. CY - Washington, DC, USA AN - OPUS4-29031 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Hoffmann, Katrin A1 - Würth, Christian A1 - Pauli, Jutta A1 - Mathejczyk, J. A1 - Alves, F. A1 - Resch-Genger, Ute T1 - NIR-Fluorescent Nano- and Micrometer-sized Particles for Biomedical and Diagnostical Applications T2 - Faraday Discussion, University of Edinburgh CY - Edinburgh, Scotland DA - 2010-09-06 PY - 2010 AN - OPUS4-22073 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Behnke, Thomas A1 - Mathejczyk, J.E. A1 - Brehm, Robert A1 - Würth, Christian A1 - Gomes, F.R. A1 - Dullin, C. A1 - Napp, J. A1 - Alves, F. A1 - Resch-Genger, Ute T1 - Target-specific nanoparticles containing a broad band emissive NIR dye for the sensitive detection and characterization of tumor development N2 - Current optical probes including engineered nanoparticles (NPs) are constructed from near infrared (NIR)-emissive organic dyes with narrow absorption and emission bands and small Stokes shifts prone to aggregation-induced self-quenching. Here, we present the new asymmetric cyanine Itrybe with broad, almost environment-insensitive absorption and emission bands in the diagnostic window, offering a unique flexibility of the choice of excitation and detection wavelengths compared to common NIR dyes. This strongly emissive dye was spectroscopically studied in different solvents and encapsulated into differently sized (15, 25, 100 nm) amino-modified polystyrene NPs (PSNPs) via a one-step staining procedure. As proof-of-concept for its potential for pre-/clinical imaging applications, Itrybe-loaded NPs were surface-functionalized with polyethylene glycol (PEG) and the tumor-targeting antibody Herceptin and their binding specificity to the tumor-specific biomarker HER2 was systematically assessed. Itrybe-loaded NPs display strong fluorescence signals in vitro and in vivo and Herceptin-conjugated NPs bind specifically to HER2 as demonstrated in immunoassays as well as on tumor cells and sections from mouse tumor xenografts in vitro. This demonstrates that our design strategy exploiting broad band-absorbing and -emitting dyes yields versatile and bright NIR probes with a high potential for e.g. the sensitive detection and characterization of tumor development and progression. KW - Nanoparticle KW - Fluorescence KW - In vitro test KW - In vivo test KW - Surface modification KW - Cytotoxicity PY - 2013 DO - https://doi.org/10.1016/j.biomaterials.2012.09.028 SN - 0142-9612 VL - 34 IS - 1 SP - 160 EP - 170 PB - Elsevier CY - Oxford AN - OPUS4-26877 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Laux, M. A1 - Hoffmann, Katrin A1 - Resch-Genger, Ute A1 - Peters, S. A1 - Haueisen, J. A1 - Klemm, M. T1 - Simple Approaches to Fluorescence Lifetime Standards Using Dye-Quencher Pairs T2 - BMT 2012 - Tagung der Biomedizinischen Technik CY - Jena, Germany DA - 2012-09-16 PY - 2012 AN - OPUS4-27071 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Behnke, Thomas A1 - Würth, Christian A1 - Laux, Eva-Maria A1 - Hoffmann, Katrin A1 - Resch-Genger, Ute T1 - Simple strategies towards bright polymer particles via one-step staining procedures N2 - In order to develop simple and versatile procedures for the preparation of red emissive particles, various one-step swelling procedures for the loading of fluorophores into nanometer- and micrometer-sized polystyrene particles were systematically assessed. Parameters studied for model dyes from common dye classes include the composition of the swelling medium, dye charge and polarity, dye concentration, and particle surface chemistry. The dye loading procedures were compared based upon the efficiency of dye incorporation, fluorescence intensity, and colloidal stability of the resulting particles as well as the absence of dye leaking as determined by absorption and fluorescence spectroscopy, flow cytometry, and measurements of zeta potentials. In addition, for the first time, the influence of the amount of incorporated dye on the absolute fluorescence quantum yield and brightness of the fluorescent particles was investigated for selected chromophores in differently sized particles using a custom-made calibrated integrating sphere setup. Our results demonstrate the general suitability of these one-step loading procedures for efficient particle staining with neutral, zwitterionic, and charged fluorophores like oxazines, coumarines, squaraines, xanthenes, and cyanines emitting in the visible and near infrared. Dye polarity was identified as a suitable tool to estimate the loading efficiency of fluorophores into these polymer particles. KW - Fluorescence KW - Polystyrene KW - Particles KW - Encapsulation KW - Quantum yield KW - Zeta potential KW - Method KW - Label KW - Particle KW - Polymer KW - Absolute fluorescence quantum yield KW - Fluorophore KW - Dye content KW - Surface groups KW - Size KW - Brightness PY - 2012 DO - https://doi.org/10.1016/j.dyepig.2012.01.021 SN - 0143-7208 SN - 1873-3743 VL - 94 IS - 2 SP - 247 EP - 257 PB - Elsevier Ltd. CY - Kidlington AN - OPUS4-25481 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Laux, Eva-Maria A1 - Behnke, Thomas A1 - Hoffmann, Katrin A1 - Resch-Genger, Ute T1 - Keeping particles brilliant - simple methods for the determination of the dye content of fluorophore-loaded polymeric particles N2 - One of the most active research areas in the life and material sciences is the design and synthesis of fluorescent nano- and micrometre sized particles for applications e.g. as labels, sensor systems, and platforms for fluorescence assays or barcoding materials. The reliable and reproducible fabrication of such particles as well as many applications require accurate, simple, and versatile procedures for the determination of the dye content per particle which affects e.g. the brightness of these materials and their surface charge and thus, colloidal stability. Here, four fast and inexpensive spectroscopic methods for the quantification of the fluorophore content of beads are presented and compared for nanometre- and micrometre sized polystyrene particles loaded or labeled with commercial fluorophores, differing in dye class, charge, and hydrophilicity. This included the determination of the amount of incorporated dye from absorption spectra of bead suspensions, via dissolving of the polymer matrix, via extraction of the polymer matrix, and from the supernatant of the swelling solution or reaction mixture. Method validation was performed with a sulfur-containing dye and elemental analysis. Based upon this method comparison and the accomplishable uncertainties, two reliable strategies for particle characterization and bead process control are identified that can be easily extended to other materials. PY - 2012 DO - https://doi.org/10.1039/c2ay05822g SN - 1759-9660 SN - 1759-9679 VL - 4 IS - 6 SP - 1759 EP - 1768 PB - RSC Publ. CY - Cambridge AN - OPUS4-26051 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hoffmann, Katrin A1 - Behnke, Thomas A1 - Grabolle, Markus A1 - Resch-Genger, Ute T1 - Nanoparticle-encapsulated vis- and NIR-emissive fluorophores with different fluorescence decay kinetics for lifetime multiplexing N2 - Bioanalytical, clinical, and security applications increasingly require simple, efficient, and versatile strategies to measure an ever increasing number of analytes or events in parallel in a broad variety of detection formats as well as in conjunction with chromatographic separation techniques or flow cytometry. An attractive alternative to common optical multiplexing and encoding methods utilizing spectral multiplexing/color encoding and intensity encoding is lifetime multiplexing, which relies on the discrimination between different fluorescent reporters based on their fluorescence decay kinetics. Here, we propose a platform of surface-functionalizable polymeric nanoparticles stained with fluorophores differing in their fluorescence lifetimes as a new multiplexing and encoding approach. Proof-of-concept measurements with different sets of lifetime-encoded polystyrene nanoparticles are presented, obtained via staining of preformed particles with visible (vis)- and near-infrared (NIR)-emissive organic dyes, which display very similar absorption and emission spectra to enable excitation and detection at the same wavelengths, yet sufficiently different fluorescence decay kinetics in suspension, thereby minimizing instrumentation costs. Data analysis was performed with a linear combination approach in the lifetime domain. Our results and first cell experiments with these reporter sets underline the suitability of our multiplexing strategy for the discrimination between and the quantification of different labels. This simple and versatile concept can be extended to all types of fluorophores, thereby expanding the accessible time scale, and can be used, e.g., for the design of labels and targeted probes for fluorescence assays and molecular imaging, cellular imaging studies, and barcoding applications, also in conjunction with spectral and intensity encoding. KW - Fluorescent label KW - Multiplexing KW - Optical KW - encoding KW - Lifetime multiplexing KW - Fluorescence lifetime imaging FLIM KW - Nanoparticles PY - 2014 DO - https://doi.org/10.1007/s00216-013-7597-3 SN - 1618-2642 SN - 1618-2650 VL - 406 IS - 14 SP - 3315 EP - 3322 PB - Springer CY - Berlin AN - OPUS4-30077 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Ralf A1 - Weigert, Florian A1 - Lesnyak, V. A1 - Leubner, S. A1 - Lorenz, T. A1 - Behnke, Thomas A1 - Dubavik, A. A1 - Joswig, J.-O. A1 - Resch-Genger, Ute A1 - Gaponik, N. A1 - Eychmüller, A. ED - Resch-Genger, Ute ED - Schneider, Ralf T1 - pH and concentration dependence of the optical properties of thiol-capped CdTe nanocrystals in water and D2O N2 - The optical properties of semiconductor nanocrystals (SC NCs) are largely controlled by their size and surface chemistry, i.e., the chemical composition and thickness of inorganic passivation shells and the chemical nature and number of surface ligands as well as the strength of their bonds to surface atoms. The latter is particularly important for CdTe NCs, which – together with alloyed CdₓHg₁₋ₓTe – are the only SC NCs that can be prepared in water in high quality without the need for an additional inorganic passivation shell. Aiming at a better understanding of the role of stabilizing ligands for the control of the application-relevant fluorescence features of SC NCs, we assessed the influence of two of the most commonly used monodentate thiol ligands, thioglycolic acid (TGA) and mercaptopropionic acid (MPA), on the colloidal stability, photoluminescence (PL) quantum yield (QY), and PL decay behavior of a set of CdTe NC colloids. As an indirect measure for the strength of the coordinative bond of the ligands to SC NC surface atoms, the influence of the pH (pD) and the concentration on the PL properties of these colloids was examined in water and D₂O and compared to the results from previous dilution studies with a set of thiol-capped Cd₁₋ₓHgₓTe SC NCs in D₂O. As a prerequisite for these studies, the number of surface ligands was determined photometrically at different steps of purification after SC NC synthesis with Ellman’s test. Our results demonstrate ligand control of the pH-dependent PL of these SC NCs, with MPA-stabilized CdTe NCs being less prone to luminescence quenching than TGA-capped ones. For both types of CdTe colloids, ligand desorption is more pronounced in H₂O compared to D₂O, underlining also the role of hydrogen bonding and solvent molecules. KW - Quantum dots KW - Fluorescence KW - Ligand analysis KW - Nano particles PY - 2016 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-371253 DO - https://doi.org/10.1039/c6cp03123d VL - 18 IS - 28 SP - 19083 EP - 19092 PB - RSC CY - Cambridge AN - OPUS4-37125 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Quevedo, Pablo A1 - Moser, Marko A1 - Würth, Christian A1 - Resch-Genger, Ute T1 - Design and Characterization of Streptavidin Modified Fluorescent Nanoparticles T2 - Molekulare Bildgebung 2014 CY - Jena, Germany DA - 2014-10-08 PY - 2014 AN - OPUS4-31613 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Resch-Genger, Ute A1 - Hoffmann, Katrin A1 - Würth, Christian A1 - Behnke, Thomas A1 - Hoffmann, Angelika A1 - Pfeifer, Dietmar A1 - Engel, A. T1 - The toolbox of fluorescence standards: Flexible calibration tools for the standardization of fluorescence-based measurements N2 - To improve the reliability of fluorescence data in the life and material sciences and to enable accreditation of fluorescence techniques, standardization concepts are required that guarantee and improve the comparability of fluorescence measurements. At the core of such concepts are simple and evaluated fluorescence standards for the consideration of instrument-specific spectral and intensity distortions of measured signals and for instrument performance validation (IPV). Similarly in need are fluorescence intensity standards for the quantification from measured intensities and for signal referencing, thereby accounting for excitation light-induced intensity fluctuations. These standards should be preferably certified, especially for use in regulated areas like medical diagnostics. This encouraged us to develop liquid and solid standards for different fluorescence parameters and techniques for use under routine measurement conditions in different formates. Special emphasis was dedicated to the determination and control of the spectral responsivity of detection systems, wavelength accuracy, homogeneity of illumination, and intensity referencing for e.g. spectrofluorometers, fluorescence sensors and confocal laser scanning fluorescence microscopes. Here, we will present design concepts and examples for mono- and multifunctional fluorescence standards that provide traceability to radiometric units and present a first step towards a toolbox of standards. KW - Fluorescence KW - Fluorescence standard KW - Calibration tool KW - Spectral fluorescence standard KW - Intensity standard KW - Instrument performance validation KW - Quality assurance KW - Traceability KW - Glass KW - Liquid standard PY - 2010 DO - https://doi.org/10.1117/12.853133 SN - 0277-786X SN - 0038-7355 SN - 0361-0748 VL - 7666 IS - 76661J SP - 1 EP - 12 PB - Soc. CY - Redondo Beach, Calif. [u.a.] AN - OPUS4-21595 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Behnke, Thomas A1 - Hoffmann, Katrin A1 - Resch-Genger, Ute T1 - Preparation, Characterization, and Bioanalytical Applications of Fluorescent Nano- and Micrometer-sized Particles T2 - Tag der Chemie CY - Berlin, Germany DA - 2010-06-16 PY - 2010 AN - OPUS4-21550 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Resch-Genger, Ute A1 - Behnke, Thomas A1 - Brehm, Robert A1 - Grabolle, Markus A1 - Hennig, Andreas A1 - Hoffmann, Angelika A1 - Hoffmann, Katrin A1 - Linck, Lena A1 - Lochmann, Cornelia A1 - Pauli, Jutta A1 - Spieles, Monika A1 - Würth, Christian T1 - Funktionelle Chromophor-Systeme, innovative Validierungskonzepte und rückführbare Standards für die fluoreszenzbasierte multiparametrische Bioanalytik N2 - Unter dem Motto „Innovation und Qualitätssicherung in der (Bio)Analytik“ werden in der Arbeitsgruppe Fluoreszenzspektroskopie der BAM, Bundesanstalt für Materialforschung und -prüfung, funktionelle Chromophor-Systeme, einfache Signalverstärkungs- und Multiplexingstrategien sowie innovative Validierungs- und rückführbare Standardisierungskonzepte für verschiedene fluorometrische Messgrößen und Methoden entwickelt. Im Mittelpunkt stehen dabei molekulare Fluorophore, Nanokristalle mit größenabhängigen optischen Eigenschaften (sogenannte Quantenpunkte, QDs) und fluoreszierende Partikel variabler Größe sowie Sonden und Sensormoleküle für neutrale und ionische Analyte und für die Charakterisierung von funktionellen Gruppen. Dabei erfolgen auch methodische Entwicklungen für die Fluoreszenzspektroskopie, die Fluoreszenzmikroskopie, die Milcrofluorometrie, die Sensorik und die Mikroarraytechnologie. Ziele sind u. a. das Design und die Untersuchung von multiplexfähigen selektiven und sensitiven Sonden für die Biomarkeranalytik, die Entwicklung von Methoden zur Charakterisierung der signalrelevanten Eigenschaften dieser Chromophor-Systeme und zur Charakterisierung von funktionellen Gruppen an Oberflächen und ihre Validierung sowie die Entwicklung und Bereitstellung von formatadaptierbaren, flexibel ersetzbaren Standards für die fluoreszenzbasierte Multiparameteranalytik. T2 - 5. Senftenberger Innovationsforum Multiparameteranalytik CY - Senftenberg, Deutschland DA - 10.03.2011 KW - Multiparametric KW - Multiplexing KW - Fluorescence KW - Nanoparticles KW - NIR dyes KW - Surface analysis KW - Quantum yield KW - Quantum dot KW - Lifetime PY - 2011 SP - 86 EP - 108 CY - Senftenberg AN - OPUS4-23635 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hoffmann, Katrin A1 - Behnke, Thomas A1 - Drescher, Daniela A1 - Kneipp, Janina A1 - Resch-Genger, Ute T1 - Lifetime-based discrimination between spectrally matching vis and NIR emitting particle labels and probes N2 - Increasing the information content from bioassays which requires robust and efficient strategies for the detection of multiple analytes or targets in a single measurement is an important field of research, especially in the context of meeting current security and health concerns. An attractive alternative to spectral multiplexing, which relies on fluorescent labels excitable at the same wavelength, yet sufficiently differing in their emission spectra or color presents lifetime multiplexing. For this purpose, we recently introduced a new strategy based on 'pattern-matching' in the lifetime domain, which was exemplary exploited for the discrimination between organic dyes and quantum dot labels revealing multi-exponential decay kinetics and allowed quantification of these labels. Meanwhile, we have succeeded in extending this lifetime multiplexing approach to nanometer-sized particle labels and probes absorbing and emitting in the visible (vis) and near-infrared (NIR) spectral region. Here, we present a first proof-of-principle of this approach for a pair of NIR-fluorescent particles. Each particle is loaded with a single organic dye chosen to display very similar absorption and emission spectra, yet different fluorescence decay kinetics. Examples for the lifetime-based distinction between pairs of these fluorescent nanoparticles in solution and in cells are presented. The results underline the potential of fluorescenc lifetime multiplexing in life science and bioanalysis. KW - Fluorescence KW - Fluorescence lifetime imaging microscopy KW - FLIM KW - Lifetime Multiplexing KW - Particle Label KW - Near-infrared KW - NIR KW - Cell imaging KW - Nanoparticles PY - 2011 DO - https://doi.org/10.1117/12.881442 SN - 1605-7422 VL - 7905 SP - 79051F-1 EP - 79051F-9 PB - SPIE, The International Society for Optical Engineering CY - Bellingham, Wash. AN - OPUS4-23637 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -