TY - JOUR A1 - Würth, Christian A1 - Pauli, Jutta A1 - Lochmann, Cornelia A1 - Spieles, Monika A1 - Resch-Genger, Ute T1 - Integrating sphere setup for the traceable measurement of absolute photoluminescence quantum yields in the near infrared JF - Analytical chemistry N2 - There is an increasing interest in chromophores absorbing and emitting in the near-infrared (NIR) spectral region, e.g., for applications as fluorescent reporters for optical imaging techniques and hence, in reliable methods for the characterization of their signal-relevant properties like the fluorescence quantum yield (Φf) and brightness. The lack of well established Φf standards for the NIR region in conjunction with the need for accurate Φf measurements in transparent and scattering media encouraged us to built up an integrating sphere setup for spectrally resolved measurements of absolute fluorescence traceable to radiometric scales. Here, we present the design of this setup and its characterization and validation including an uncertainty budget for the determination of absolute Φf in the visible and NIR. To provide the basis for better measurements of Φf in the spectral window from ca. 600 to 1000 nm used, e.g., for optical imaging, the absolute Φf of a set of NIR chromophores covering this spectral region are measured and compared to relative values obtained using rhodamine 101 as Φf standard. Additionally, the absolute Φf values of some red dyes that are among the most commonly used labels in the life sciences are presented as well as the absolute quantum yield of an optical probe for tumor imaging. KW - Fluorescence KW - Photoluminescence KW - Quantum yield KW - Dye KW - Uncertainty KW - Absolute quantum yield KW - Method comparison KW - Integrating sphere KW - NIR KW - Standards KW - Quantum yield standards PY - 2012 DO - https://doi.org/10.1021/ac2021954 SN - 0003-2700 SN - 1520-6882 VL - 84 IS - 3 SP - 1345 EP - 1352 PB - American Chemical Society CY - Washington, DC AN - OPUS4-25504 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Würth, Christian A1 - Lochmann, Cornelia A1 - Spieles, Monika A1 - Pauli, Jutta A1 - Hoffmann, Katrin A1 - Schüttrigkeit, T. A1 - Franzl, T. A1 - Resch-Genger, Ute T1 - Evaluation of a commercial integrating sphere setup for the determination of absolute photoluminescence quantum yields of dilute dye solutions JF - Applied spectroscopy N2 - The commercial availability of stand-alone setups for the determination of absolute photoluminescence quantum yields (φf) in conjunction with the increasing use of integrating sphere accessories for spectrofluorometers is expected to have a considerable influence not only on the characterization of chromophore systems for use in optical and opto-electronic devices, but also on the determination of this key parameter for (bio)analytically relevant dyes and functional luminophores. Despite the huge potential of systems measuring absolute φf values and the renewed interest in dependable data, evaluated protocols for even the most elementary case, the determination of the fluorescence quantum yield of transparent dilute solutions of small organic dyes with integrating sphere methods, are still missing. This encouraged us to evaluate the performance and sources of uncertainty of a simple commercial integrating sphere setup with dilute solutions of two of the best characterized fluorescence quantum yield standards, quinine sulfate dihydrate and rhodamine 101, strongly differing in spectral overlap between absorption and emission. Special attention is dedicated to illustrate common pitfalls of this approach, thereby deriving simple procedures to minimize measurement uncertainties and improve the comparability of data for the broad community of users of fluorescence techniques. KW - Lifetime KW - Fluorescence KW - Luminescence KW - Quantum yield KW - Quantum efficiency KW - Integrating sphere KW - Reabsorption KW - Rhodamine 101 KW - Quinine sulfate dihydrate KW - Method KW - Photoluminescence KW - Standard KW - Emission KW - Spectral correction KW - Excitation KW - Anisotropy PY - 2010 DO - https://doi.org/10.1366/000370210791666390 SN - 0003-7028 SN - 1943-3530 VL - 64 IS - 7 SP - 733 EP - 741 PB - Society for Applied Spectroscopy CY - Frederick, Md. AN - OPUS4-22089 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Würth, Christian A1 - Grabolle, Markus A1 - Pauli, Jutta A1 - Spieles, Monika A1 - Resch-Genger, Ute T1 - Relative and absolute determination of fluorescence quantum yields of transparent samples JF - Nature protocols N2 - Luminescence techniques are among the most widely used detection methods in the life and material sciences. At the core of these methods is an ever-increasing variety of fluorescent reporters (i.e., simple dyes, fluorescent labels, probes, sensors and switches) from different fluorophore classes ranging from small organic dyes and metal ion complexes, quantum dots and upconversion nanocrystals to differently sized fluorophore-doped or fluorophore-labeled polymeric particles. A key parameter for fluorophore comparison is the fluorescence quantum yield (Φf), which is the direct measure for the efficiency of the conversion of absorbed light into emitted light. In this protocol, we describe procedures for relative and absolute determinations of Φf values of fluorophores in transparent solution using optical methods, and we address typical sources of uncertainty and fluorophore class-specific challenges. For relative determinations of Φf, the sample is analyzed using a conventional fluorescence spectrometer. For absolute determinations of Φf, a calibrated stand-alone integrating sphere setup is used. To reduce standard-related uncertainties for relative measurements, we introduce a series of eight candidate quantum yield standards for the wavelength region of ~350–950 nm, which we have assessed with commercial and custom-designed instrumentation. With these protocols and standards, uncertainties of 5–10% can be achieved within 2 h. PY - 2013 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-294202 DO - https://doi.org/10.1038/nprot.2013.087 SN - 1754-2189 SN - 1750-2799 VL - 8 IS - 8 SP - 1535 EP - 1550 PB - Nature Publishing Group CY - Basingstoke AN - OPUS4-29420 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Resch-Genger, Ute A1 - Würth, Christian A1 - Pauli, Jutta A1 - Weigert, Florian T1 - Quantitative optical-spectroscopic characterization of luminescent nanomaterials - Photoluminescence Quantum Yields N2 - Accurate and quantitative photoluminescence measurements are mandatory for the comparison of different emitter classes and the rational design of the next generation of molecular and nanoscale reporters as well as for most applications relying on their luminescence features in the life and material sciences and nanobiophotonics. In the following, procedures for the determination of the spectroscopic key parameter photoluminescence quantum yield, i.e., the number of emitted per absorbed photons, in the UV/vis/NIR/SWIR are presented including pitfalls and achievable uncertainties and material-specific effects related to certain emitter classes are addressed. Special emphasis is dedicated to luminescent nanocrystals. T2 - International Workshop on "Emerging Nanomaterials for Displays and SSL" CY - Dresden, Germany DA - 11.11.2021 KW - Nano KW - Nanomaterial KW - Nanocrystal KW - Semiconductor quantum dot KW - Lanthanide KW - Photoluminescence KW - Quantum yield KW - Photophysics KW - Lifetime KW - Surface chemistry KW - Brightness KW - NIR KW - SWIR KW - Method KW - Uncertainty KW - Rreference material PY - 2021 AN - OPUS4-53783 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Pauli, Jutta A1 - Würth, Christian A1 - Güttler, Arne A1 - Resch-Genger, Ute T1 - Reliable Determination of the Signal-Relevant Spectroscopic Key Characteristics of Luminescent Reporters and Optical Probes for Imaging in the vis/NIR/SWIR N2 - Introduction. Comparing different emitter classes and rationally designing the next generation of molecular and nanoscale probes for bioimaging applications require accurate and quantitative methods for the measurement of the key parameter photoluminescence quantum yield f.1 f equals the number of emitted per number of absorbed photons. This is particularly relevant for increasingly used fluorescence imaging in the short wave-infrared region (SWIR) ≥ 900 nm providing deeper penetration depths, a better image resolution, and an improved signal-to-noise or tumor-to-background ratio.2, 3 However, spectroscopic measurements in the SWIR are more challenging and require specific calibrations and standards. T2 - EMIM 2021 CY - Göttingen, Germany DA - 24.08.2021 KW - Fluorescence KW - Optical probe KW - Dye KW - Photophysics KW - Quantum yield KW - Mechanism KW - NIR KW - SWIR KW - Imaging KW - Reference material KW - Reliability KW - Nano KW - Particle KW - Method KW - Quality assurance PY - 2021 AN - OPUS4-53233 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Würth, Christian A1 - Grabolle, Markus A1 - Pauli, Jutta A1 - Spieles, Monika A1 - Resch-Genger, Ute T1 - Comparison of methods and achievable uncertainties for the relative and absolute measurement of photoluminescence quantum yields JF - Analytical chemistry N2 - The photoluminescence quantum yield (Φf) that presents a direct measure for the efficiency of the conversion of absorbed photons into emitted photons is one of the spectroscopic key parameters of functional fluorophores. It determines the suitability of such materials for applications in, for example, (bio)analysis, biosensing, and fluorescence imaging as well as as active components in optical devices. The reborn interest in accurate Φf measurements in conjunction with the controversial reliability of reported Φf values of many common organic dyes encouraged us to compare two relative and one absolute fluorometric method for the determination of the fluorescence quantum yields of quinine sulfate dihydrate, coumarin 153, fluorescein, rhodamine 6G, and rhodamine 101. The relative methods include the use of a chain of Φf transfer standards consisting of several 'standard dye' versus 'reference dye' pairs linked to a golden Φf standard that covers the ultraviolet and visible spectral region, and the use of different excitation wavelengths for standard and sample, respectively. Based upon these measurements and the calibration of the instruments employed, complete uncertainty budgets for the resulting Φf values are derived for each method, thereby providing evaluated standard operation procedures for Φf measurements and, simultaneously, a set of assessed Φf standards. KW - Fluorescence KW - Photoluminescence KW - Quantum yield KW - Dye KW - Uncertainty KW - Absolute quantum yield KW - Method comparison KW - Integrating sphere PY - 2011 DO - https://doi.org/10.1021/ac2000303 SN - 0003-2700 SN - 1520-6882 VL - 83 IS - 9 SP - 3431 EP - 3439 PB - American Chemical Society CY - Washington, DC AN - OPUS4-23632 LA - eng 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 T2 - 5. Senftenberger Innovationsforum Multiparameteranalytik (Proceedings) 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 - Hatami, Soheil A1 - Würth, Christian A1 - Kaiser, Martin A1 - Leubner, S. A1 - Gabriel, S. A1 - Bahrig, L. A1 - Lesnyak, V. A1 - Pauli, Jutta A1 - Gaponik, N. A1 - Eychmüller, A. A1 - Resch-Genger, Ute T1 - Absolute photoluminescence quantum yields of IR26 and IR-emissive Cd1-xHgxTe and PbS quantum dots - method- and material-inherent challenges JF - Nanoscale N2 - Bright emitters with photoluminescence in the spectral region of 800–1600 nm are increasingly important as optical reporters for molecular imaging, sensing, and telecommunication and as active components in electrooptical and photovoltaic devices. Their rational design is directly linked to suitable methods for the characterization of their signal-relevant properties, especially their photoluminescence quantum yield (Φf). Aiming at the development of bright semiconductor nanocrystals with emission >1000 nm, we designed a new NIR/IR integrating sphere setup for the wavelength region of 600–1600 nm. We assessed the performance of this setup by acquiring the corrected emission spectra and Φf of the organic dyes Itrybe, IR140, and IR26 and several infrared (IR)-emissive Cd1-xHgxTe and PbS semiconductor nanocrystals and comparing them to data obtained with two independently calibrated fluorescence instruments absolutely or relative to previously evaluated reference dyes. Our results highlight special challenges of photoluminescence studies in the IR ranging from solvent absorption to the lack of spectral and intensity standards together with quantum dot-specific challenges like photobrightening and photodarkening and the size-dependent air stability and photostability of differently sized oleate-capped PbS colloids. These effects can be representative of lead chalcogenides. Moreover, we redetermined the Φf of IR26, the most frequently used IR reference dye, to 1.1 × 10-3 in 1,2-dichloroethane DCE with a thorough sample reabsorption and solvent absorption correction. Our results indicate the need for a critical reevaluation of Φf values of IR-emissive nanomaterials and offer guidelines for improved Φf measurements. PY - 2015 DO - https://doi.org/10.1039/c4nr04608k SN - 2040-3364 SN - 2040-3372 VL - 7 IS - 1 SP - 133 EP - 143 PB - RSC Publ. CY - Cambridge AN - OPUS4-32408 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pauli, Jutta A1 - Hoffmann, Katrin A1 - Würth, Christian A1 - Behnke, Thomas A1 - Resch-Genger, Ute T1 - Standardization of fluorescence measurements in the UV/vis/NIR/IR JF - Standardization of Fluorescence Measurements in the UV/vis/NIR/IR N2 - Photoluminescence techniques are amongst the most widely used Tools in the life sciences, with new and exciting applications in medical diagnostics and molecular Imaging continuously emerging. Advantages include their comparative ease of use, unique sensitivity, non-invasive character, and potential for Multiplexing, remote sensing, and miniaturization. General drawbacks are, however, signals, that contain unwanted wavelength- and polarization contributions from Instrument-dependent effects, which are also time-dependent due to aging of Instrument-components, and difficulties to measure absolute flourescence entensities. Moreover, scattering Systems require Special measurement geometries and the interest in new optical Reporters with Emission > 1000 nm strategies for reliable measurements in the second diagnostic for the comparison of material Performance and the rational designg of new flourophores with improved properties. Here, we present strategies to versatile method-adaptable liquid and solid flourescence Standards for different flourescence paramters including traceable Instrument calibration procedures and the design of integrating spere setups for the absolute measurements of emission spectra and Quantum yields in the wavelength Region of 350 to 1600 nm. Examples are multi-Emitter glasses, spectral flourescence Standards, and quantum yield Standards for the UV/vis/NIR. T2 - Conference on Molecular-Guided Surgery - Molecules, Devices, and Applications III CY - San Francisco, CA, USA DA - 28.01.2017 KW - Fluorescence KW - Reference material KW - Standard KW - Calibration KW - Nanoparticle KW - Absolute flourometry KW - Integrating sphere spectroscopy KW - NIR KW - IR KW - Quantum yield standard KW - Emission standards PY - 2017 SN - 978-1-5106-0539-8 DO - https://doi.org/10.1117/12.2255728 SN - 0277-786X VL - 10049 SP - 1 PB - Proceedings of SPIE AN - OPUS4-41783 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -