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Fluorescence techniques such as fluorescence spectroscopy, microfluorometry, and fluorescence microscopy, providing spectral, intensity, polarization, and lifetime information, are amongst the most broadly utilized analytical methods in the life and materials sciences. However, the measured fluorescence data contain sample- and instrument-specific contributions, which hamper their comparability across instruments and laboratories. Comparable, instrument-independent fluorescence data require the determination of the fluorescence instrument’s wavelength-dependent spectral responsivity, also termed emission correction curve, for the same instrument settings as those used for the fluorescence measurements as a prerequisite for the subsequent correction of the measured instrument-specific data. Such a spectral correction is essential for the performance comparison of different fluorescent labels and reporters, quantitative fluorescence measurements, the determination of the fluorescence quantum yield, and the spectroscopic measure for the fluorescence efficiency of a fluorophore. Simple-to-use tools for obtaining emission correction curves are chromophore-based reference materials (RMs), referred to as fluorescence standards, with precisely known, preferably certified instrument-independent fluorescence spectra. However, for the increasingly used near-infrared (NIR) wavelength region >700 nm, at present, no spectral fluorescence standards are available. To close this gap, we developed two novel spectral fluorescence standards, BAM F007 and BAM-F009, with broad emission bands from about 580 to 940 nm in ethanolic solution. These liquid fluorescence standards currently under certification, which will be released in 2025, will expand the wavelength range of the already available certified Calibration Kit BAM F001b-F005b from about 300–730 to 940 nm. In this research article, we will detail the criteria utilized for dye and matrix selection and the homogeneity and stability tests accompanying dye certification as well as the calculation of the wavelength-dependent uncertainty budgets of the emission spectra BAM F007 and BAM-F009, determined with the traceably calibrated BAM reference spectrofluorometer. These fluorescence standards can provide the basis for comparable fluorescence measurements in the ultraviolet, visible, and NIR for the fluorescence community.
Luminescence techniques are amongst the most commonly used analytical methods in the life and material sciences due to their high sensitivity and non-destructive and multiparametric character. Photoluminescence signals are, however, affected by wavelength-, polarization-, and time-dependent instrument specific effect and the compound-specific photoluminescence quantum yield. The former hamper the comparability of fluorescence measurements, while the relative determination of the latter requires suitable quantum yield standards with well-known photoluminescence quantum yields (QY). For the simple correction of instrument specific effects in the wavelength region of 300 nm to 950 nm, the set of the five certified spectral fluorescence standards BAM-F001 – BAM-F005, has been extended to the NIR range by including two new fluorescence standards currently under certification. For the reliable and accurate determination of QY which is the key performance parameter for the comparison of different luminophores, we certified a set of 12 quantum yield standards, which absorb and emit in the wavelength range from 300 nm to 1000 nm.
We report the synthesis of a fluorescent polycyclic aromatic hydrocarbon dye with a “symmetry-broken” core, derived from the related hexa-peri-benzocoronene (HBC) core with fluoranthene subunit. The fluorophore is composed of a pure Carbon skeleton without heteroatoms and exhibits remarkable photo luminescence properties with a photoluminescence Quantum yield (PLQY) of up to 67% in toluene, exceeding that of theparent HBC by a factor of 30. The single crystal X-ray structure reveals the distorted polycyclic aromatic hydrocarbon structure, which is responsible for the optoelectronic properties, as supported by density functional theory calculations. We show that the new fluorescent dye can be readily used for the fabrication of organic light-emitting diodes (OLED) without extensive optimization, whereby solubility in a variety of solvents and successful film formation are decisive.
For a series of 500 μm-thick polyurethane films containing different concentrations of luminescent and scattering YAG:Ce microparticles, we systematically explored and quantified pitfalls of absolute measurements of photoluminescence quantum yields (Φf) for often employed integrating sphere (IS) geometries, where the sample is placed either on a sample holder at the bottom of the IS surface or mounted in the IS center. Thereby, the influence of detection and illumination geometry and sample position was examined using blanks with various scattering properties for measuring the number of photons absorbed by the sample. Our results reveal that (i) setup configurations where the scattering sample is mounted in the IS center and (ii) transparent blanks can introduce systematic errors in absolute Φf measurements.
For strongly scattering, luminescent samples, this can result in either an under- or overestimation of the absorbed photon flux and hence an under- or overestimation of Φf. The size of these uncertainties depends on the scattering properties of the sample and instrument parameters, such as sample position, IS size, wavelength-dependent reflectivity of the IS surface coating, and port configuration. For accurate and reliable absolute Φf measurements, we recommend (i) a blank with scattering properties closely matching those of the sample to realize similar distributions of the diffusely scattered excitation photons within the IS, and (ii) a sufficiently high sample absorption at the excitation wavelength. For IS setups with center-mounted samples, measurement geometries should be utilized that prevent the loss of excitation photons by reflections from the sample out of the IS.
The size and shape of photoluminescence signals is affected by wavelength-, polarization-, and time-dependent instrumentspecific contributions and the compound- and environment-specific photoluminescence quantum yield. The former hamper the comparability of fluorescence measurements performed on different measuring devices. The commonly relatively done determination of the performance parameter requires suitable quantum yield standards with well-known. The performance of such measurements is, e.g., described in the written standard IEC 62607 currently revised.
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.
New Reference Materials for Quantification and Standardization of Fluorescence-based Measurements
(2022)
Luminescence techniques are amongst the most commonly used analytical methods in the life and material sciences due to their high sensitivity and non-destructive and multiparametric character. Photoluminescence signals are, however, affected by wavelength-, polarization-, and time-dependent instrument specific effects and the compound-specific photoluminescence quantum yield. The former hamper the comparability of fluorescence measurements, while the relative determination of the latter requires suitable quantum yield standards with well-known photoluminescence quantum yields. For the simple correction of instrument specific effects in the wavelength region of 300 nm to 950 nm, the set of the five certified spectral fluorescence standards (CRM) BAM-F001 – BAM-F005, has been extended to the NIR range by including two new fluorescence standards currently under certification.
Photoluminescence Quantum Yields of Luminescent Nanocrystals and Particles in the UV/vis/NIR/SWIR
(2023)
The rational design of functional luminescent materials such as semiconductor quantum dots and lanthanide-based upconversion nanoparticles, all photophysical and mechanistic studies, and the comparison of different emitters require accurate and quantitative photoluminescence measurements. Particularly the reliable determination of the key performance parameter photoluminescence quantum yield (f), the number of emitted per absorbed photons, and the brightness are of special importance for luminescence applications in the life and material sciences and nano(bio)photonics.[1] In this context, examples for absolute measurements of the photoluminescence quantum yields of UV/vis/NIR/SWIR emissive semiconductor quantum dots and rods, made from different materials, and spectrally shifting lanthanide upconversion nanocrystals with different surface chemistries in transparent matrices are presented including excitation wavelength and power density dependent studies utilizing integration sphere spectroscopy.[2,3] In addition, procedures for the absolute determination of the photoluminescence quantum yields of scattering dispersions of larger size quantum rods and differently sized inorganic particles have been developed as well as procedures for the characterization of solid luminescent nanomaterials such as different perovskites and YAG:Cer converter materials.[4] Thereby, challenges and pitfalls of f measurements in different wavelength regions including the SWIR and material-specific effects related to certain emitter classes are addressed, achievable uncertainties are quantified, and relative and absolute measurements of photoluminescence quantum yield measurements are compared to underline limitations of the former approach. Finally, a set of novel UV/vis/NIR quantum yield standards is presented including their certification with a complete uncertainty budget.[5]
Organic fluorophores, particularly stimuli-responsive molecules, are very interesting for biological and material sciences applications, but frequently limited by aggregation- and rotation-caused photoluminescence quenching. A series of easily accessible bipyridinium fluorophores, whose emission is quenched by a twisted intramolecular charge-transfer (TICT) mechanism, is reported.
Encapsulation in a cucurbit[7]uril host gave a 1:1 complex exhibiting a moderate emission increase due to destabilization of the TICT state inside the apolar cucurbituril cavity. A much stronger fluorescence enhancement is observed in 2:2 complexes with the larger cucurbit[8]uril, which is caused by additional conformational restriction of rotations around the aryl/aryl bonds. Because the cucurbituril complexes are pH switchable, this system represents an efficient supramolecular ON/OFF fluorescence switch.
Diaminodicyanoquinones: Fluorescent dyes with high dipole moments and electron-acceptor properties
(2019)
Fluorescent dyes are applied in various fields of research,includingsolarcellsandlight-emittingdevices,andas reporters for assays and bioimaging studies.Fluorescent dyes with an added high dipole moment pave the way to nonlinear optics and polarity sensitivity.Redox activity makes it possible to switch the moleculeQsphotophysical properties.Diaminodicyanoquinone derivatives possess high dipole moments,yet only lowfluorescence quantum yields,and have therefore been neglected as fluorescent dyes.Here we investigate the fluorescencepropertiesofdiaminodicyanoquinonesusingacombined theoretical and experimental approach and derive molecules with afluorescence quantum yield exceeding 90%. The diaminodicyanoquinone core moiety provides chemical versatility and can be integrated into novel molecular architectures with unique photophysical features.