Interlaboratory Comparison on Absolute Photoluminescence Quantum Yield Measurements of Solid Light Converting Phosphors with Three Commercial Integrating Sphere Setups.

Scattering luminescent materials dispersed in liquid and solid matrices and luminescent powders are increasingly relevant for fundamental research and industry. Examples are luminescent nano- and microparticles and phosphors of different compositions in various matrices or incorporated into ceramics with applications in energy conversion, solid-state lighting, medical diagnostics, and security barcoding. The key parameter to characterize the performance of these materials is the photoluminescence/fluorescence quantum yield (Φf), i.e., the number of emitted photons per number of absorbed photons. To identify and quantify the sources of uncertainty of absolute measurements of Φf of scattering samples, the first interlaboratory comparison (ILC) of three laboratories from academia and industry was performed by following identical measurement protocols. Thereby, two types of commercial stand-alone integrating sphere setups with different illumination and detection geometries were utilized for measuring the Φf of transparent and scattering dye solutions and solid phosphors, namely, YAG:Ce optoceramics of varying surface roughness, used as converter materials for blue light emitting diodes. Special emphasis was dedicated to the influence of the measurement geometry, the optical properties of the blank utilized to determine the number of photons of the incident excitation light absorbed by the sample, and the sample-specific surface roughness. While the Φf values of the liquid samples matched between instruments, Φf measurements of the optoceramics with different blanks revealed substantial differences. The ILC results underline the importance of the measurement geometry, sample position, and blank for reliable Φf data of scattering the YAG:Ce optoceramics, with the blank's optical properties accounting for uncertainties exceeding 20%.

Efficiency scale for scattering luminescent particles linked to fundamental and measurable spectroscopic properties.

Comparing the performance of molecular and nanoscale luminophores and luminescent micro- and nanoparticles and estimating achievable signal amplitudes and limits of detection requires a standardizable intensity scale. This initiated the development of the relative MESF (number of molecules of equivalent soluble fluorochromes) and ERF (equivalent reference fluorophores) scales for flow cytometry and fluorescence microscopy. Both intensity scales rely on fluorescence intensity values assigned to fluorescent calibration beads by an intensity comparison to spectrally closely matching fluorophore solutions of known concentration using a spectrofluorometer. Alternatively, the luminophore or bead brightness (B) can be determined that equals the product of the absorption cross section (σa) at the excitation wavelength (σa(λex)) and the photoluminescence quantum yield (Φpl). Thereby, an absolute scale based on fundamental and measurable spectroscopic properties can be realized which is independent of particle size, material, and luminophore staining or labeling density and considers the sensitivity of the optical properties of luminophores to their environment. Aiming for establishing such a brightness scale for light-scattering dispersions of luminescent particles with sizes exceeding a few ten nanometers, we demonstrate how the brightness of quasi-monodisperse 25 nm, 100 nm, and 1 µm sized polystyrene particles (PSP), loaded with two different dyes in varying concentrations, can be obtained with a single custom-designed integrating sphere setup that enables the absolute determination of Φpl and transmittance and diffuse reflectance measurements. The resulting Φpl, σa(λex), imaginary parts of the refractive index, and calculated B values of these samples are given in dependence of the number of incorporated dye molecule per particle. Finally, a unitless luminescence efficiency (LE) is defined allowing for the direct comparison of luminescence efficiencies of particles with different sizes.

Fluorescence Quantum Yield Standards for the UV/Visible/NIR: Development, Traceable Characterization, and Certification.

The rational design of next generation molecular and nanoscale reporters and the comparison of different emitter classes require the determination of the fluorometric key performance parameter fluorescence quantum yield (Φf), i.e., the number of emitted photons per number of absorbed photons. Main prerequisites for reliable Φf measurements, which are for transparent luminophore solutions commonly done relative to a reference, i.e., a fluorescence quantum yield standard of known Φf, are reliable and validated instrument calibration procedures to consider wavelength-, polarization-, and time-dependent instrument specific signal contributions, and sufficiently well characterized fluorescence quantum yield standards. As the standard's Φf value directly contributes to the calculation of the sample's Φf, its accuracy presents one of the main sources of uncertainty of relative Φf measurements. To close this gap, we developed a first set of 12 fluorescence quantum yield standards, which absorb and emit in the wavelength region of 330-1000 nm and absolutely determined their Φf values with two independently calibrated integrating sphere setups. Criteria for standard selection and the configuration of these novel fluorescence reference materials are given, and the certification procedure is presented including homogeneity and stability studies and the calculation of complete uncertainty budgets for the certified Φf values. The ultimate goal is to provide the community of fluorescence users with available reference materials as a basis for an improved comparability and reliability of quantum yield data since the measurement of this spectroscopic key property is an essential part of the characterization of any new emitter.

Assessing the influence of microwave-assisted synthesis parameters and stabilizing ligands on the optical properties of AIS/ZnS quantum dots.

Luminescent semiconductor quantum dots (QDs) are frequently used in the life and material sciences as reporter for bioimaging studies and as active components in devices such as displays, light-emitting diodes, solar cells, and sensors. Increasing concerns regarding the use of toxic elements like cadmium and lead, and hazardous organic solvents during QD synthesis have meanwhile triggered the search for heavy-metal free QDs using green chemistry syntheses methods. Interesting candidates are ternary AgInS2 (AIS) QDs that exhibit broad photoluminescence (PL) bands, large effective Stokes shifts, high PL quantum yields (PL QYs), and long PL lifetimes, which are particularly beneficial for applications such as bioimaging, white light-emitting diodes, and solar concentrators. In addition, these nanomaterials can be prepared in high quality with a microwave-assisted (MW) synthesis in aqueous solution. The homogeneous heat diffusion and instant temperature rise of the MW synthesis enables a better control of QD nucleation and growth and thus increases the batch-to-batch reproducibility. In this study, we systematically explored the MW synthesis of AIS/ZnS QDs by varying parameters such as the order of reagent addition, precursor concentration, and type of stabilizing thiol ligand, and assessed their influence on the optical properties of the resulting AIS/ZnS QDs. Under optimized synthesis conditions, water-soluble AIS/ZnS QDs with a PL QY of 65% and excellent colloidal and long-term stability could be reproducible prepared.

Dopant ion concentration-dependent upconversion luminescence of cubic SrF2:Yb3+,Er3+ nanocrystals prepared by a fluorolytic sol-gel method.

A fluorolytic sol-gel method was used for the fast and simple synthesis of small cubic-phase SrF2:Yb3+,Er3+ upconversion (UC) nanocrystals (UCNC) of different composition at room temperature. Systematic studies of the crystal phase and particle size of this Yb3+,Er3+-concentration series as well as excitation power density (P)-dependent UC luminescence (UCL) spectra, UCL quantum yields (ΦUCL), and UCL decay kinetics yielded maximum UCL performance for doping amounts of Yb3+ of 13.5% and Er3+ of 1.3% in the studied doping and P-range (30-400 W cm-2). Furthermore, ΦUCL were determined to be similar to popular ß-NaYF4:Yb3+,Er3+. The relative spectral UCL distributions revealed that all UCNC show a strong red emission in the studied doping and P-range (30-400 W cm-2) and suggest that the UCL quenching pathway for unshelled cubic-phase SrF2:Yb3+,Er3+ UCNC differs from the commonly accepted population and depopulation pathways of ß-NaYF4:Yb3+,Er3+ UCNC. In SrF2:Yb3+,Er3+ UCNC the 4S3/2 → 4I13/2 transition exhibits a notably stronger sensitivity towards P and reveals increasing values for decreasing Yb3+-Yb3+ distances while the 4I9/2 → 4I15/2 transition is significantly less affected by P and energy migration facilitated UCL quenching. These results emphasize the complexity of the UC processes and the decisive role of the crystal phase and symmetry of the host lattice on the operative UCL quenching mechanism in addition to surface effects. Moreover, the room temperature UCNC synthesis enabled a systematic investigation of the influence of the calcination temperature on the crystal phase of powder-UCNC and the associated UCL properties. Calcination studies of solid UCNC of optimized doping concentration in the temperature range of 175 °C and 800 °C showed the beneficial influence of temperature-induced healing of crystal defects on UCL and the onset of a phase separation connected with the oxygenation of the lanthanide ions at elevated temperature. This further emphasizes the sensitivity of the UC process to the crystal phase and quality of the host matrix.

Tailoring the SWIR emission of gold nanoclusters by surface ligand rigidification and their application in 3D bioimaging.

The influence of solvent polarity and surface ligand rigidification on the SWIR emission profile of gold nanoclusters with an anistropic surface was investigated. A strong enhancement of the SWIR emission band at 1200 nm was observed when measuring in different local environments: in solution, in polymer composites, and in solids. SWIR in vivo imaging of mice assisted by deep learning after intravenous administration of these gold nanoclusters provides high definition pseudo-3D views of vascular blood vessels.

Metasurface Enhanced Sensitized Photon Upconversion: Toward Highly Efficient Low Power Upconversion Applications and Nanoscale E-Field Sensors.

Large-scale nanoimprinted metasurfaces based on silicon photonic crystal slabs were produced and coated with a NaYF4:Yb3+/Er3+ upconversion nanoparticle (UCNP) layer. UCNPs on these metasurfaces yield a more than 500-fold enhanced upconversion emission compared to UCNPs on planar surfaces. It is also demonstrated how the optical response of the UCNPs can be used to estimate the local field energy in the coating layer. Optical simulations using the finite element method validate the experimental results and the calculated spatial three-dimensional field energy distribution helps us to understand the emission enhancement mechanism of the UCNPs closely attached to the metasurface. In addition, we analyzed the spectral shifts of the resonances for uncoated and coated metasurfaces and metasurfaces submerged in water to enable a prediction of the optimum layer thicknesses for different excitation wavelengths, paving the way to applications such as electromagnetic field sensors or bioassays.

Time-resolved luminescence spectroscopy for monitoring the stability and dissolution behaviour of upconverting nanocrystals with different surface coatings.

We demonstrate the potential of time-resolved luminescence spectroscopy for the straightforward assessment and in situ monitoring of the stability of upconversion nanocrystals (UCNPs). Therefore, we prepared hexagonal NaYF4:Yb3+,Er3+ UCNPs with various coatings with a focus on phosphonate ligands of different valency, using different ligand exchange procedures, and studied their dissolution behaviour in phosphate-buffered saline (PBS) dispersions at 20 °C and 37 °C with various analytical methods. The amount of the released UCNPs constituting fluoride ions was quantified by potentiometry using a fluoride ion-sensitive electrode and particle disintegration was confirmed by transmission electron microscopy studies of the differently aged UCNPs. In parallel, the luminescence features of the UCNPs were measured with special emphasis on the lifetime of the sensitizer emission to demonstrate its suitability as screening parameter for UCNP stability and changes in particle composition. The excellent correlation between the changes in luminescence lifetime and fluoride concentration highlights the potential of our luminescence lifetime method for UCNP stability screening and thereby indirect monitoring of the release of potentially hazardous fluoride ions during uptake and dissolution in biological systems. Additionally, the developed in situ optical method was used to distinguish the dissolution dynamics of differently sized and differently coated UCNPs.

Efficient sub-15 nm cubic-phase core/shell upconversion nanoparticles as reporters for ensemble and single particle studies.

Single particle imaging of upconversion nanoparticles (UCNPs) has typically been realized using hexagonal (ß) phase lanthanide-doped sodium yttrium fluoride (NaYF4) materials, the upconversion luminescence (UCL) of which saturates at power densities (P) of several hundred W cm-2 under 980 nm near-infrared (NIR) excitation. Cubic (α) phase UCNPs have been mostly neglected because of their commonly observed lower UCL efficiency at comparable P in ensemble level studies. Here, we describe a set of sub-15 nm ytterbium-enriched α-NaYbF4:Er3+@CaF2 core/shell UCNPs doped with varying Er3+ concentrations (5-25%), studied over a wide P range of ∼8-105 W cm-2, which emit intense UCL even at a low P of 10 W cm-2 and also saturate at relatively low P. The highest upconversion quantum yield (ΦUC) and the highest particle brightness were obtained for an Er3+ dopant concentration of 12%, reaching the highest ΦUC of 0.77% at a saturation power density (Psat) of 110 W cm-2. These 12%Er3+-doped core/shell UCNPs were also the brightest UCNPs among this series under microscopic conditions at high P of ∼102-105 W cm-2 as demonstrated by imaging studies at the single particle level. Our results underline the potential applicability of the described sub-15 nm cubic-phase core/shell UCNPs for ensemble- and single particle-level bioimaging.

Yb,Nd,Er-doped upconversion nanoparticles: 980 nm versus 808 nm excitation.

Yb,Nd,Er-doped upconversion nanoparticles (UCNPs) have attracted considerable interest as luminescent reporters for bioimaging, sensing, energy conversion/shaping, and anticounterfeiting due to their capability to convert multiple near-infrared (NIR) photons into shorter wavelength ultraviolet, visible or NIR luminescence by successive absorption of two or more NIR photons. This enables optical measurements in complex media with very little background and high penetration depths for bioimaging. The use of Nd3+ as substitute for the commonly employed sensitizer Yb3+ or in combination with Yb3+ shifts the excitation wavelength from about 980 nm, where the absorption of water can weaken upconversion luminescence, to about 800 nm, and laser-induced local overheating effects in cells, tissue, and live animal studies can be minimized. To systematically investigate the potential of Nd3+ doping, we assessed the performance of a set of similarly sized Yb3+,Nd3+,Er3+-doped core- and core-shell UCNPs of different particle architecture in water at broadly varied excitation power densities (P) with steady state and time-resolved fluorometry for excitation at 980 nm and 808 nm. As a measure for UCNPs performance, the P-dependent upconversion quantum yield (ΦUC) and its saturation behavior were used as well as particle brightness (BUC). Based upon spectroscopic measurements at both excitation wavelengths in water and in a lipid phantom and BUC-based calculations of signal size at different penetration depths, conditions under which excitation at 808 nm is advantageous are derived and parameters for the further optimization of triple-doped UCNPs are given.

Simple Self-Referenced Luminescent pH Sensors Based on Upconversion Nanocrystals and pH-Sensitive Fluorescent BODIPY Dyes.

We present the design and fabrication of pH responsive ratiometric dual component sensor systems based on multicolor emissive upconversion nanoparticles (UCNP) and pH sensitive BODIPY dyes with tunable p Ka values embedded into a polymeric hydrogel matrix. The use of NIR excitable NaYF4:Yb3+,Tm3+ UCNPs enables background free read-out. Furthermore, the spectrally matching optical properties of the UCNPs and the dyes allow the UCNPs to serve as excitation light source for the analyte-responsive BODIPY as well as intrinsic reference. The blue upconversion luminescence (UCL) of NaYF4:Yb3+,Tm3+ UCNPs excited at 980 nm, that overlaps with the absorption of the pH-sensitive fluorophore, provides reabsorption based excitation of the dye, the spectrally distinguishable green fluorescence of which is switched ON upon protonation, preventing photoinduced electron transfer (PET) within the dye moiety, and the pH-inert red UCL act as reference. The intensities ratios of the dye's fluorescence and the analyte-inert red Tm3+ UCL correlate directly with pH, which was successfully utilized for monitoring time-dependent pH changes of a suspension of quiescent E. coli metabolizing d-glucose.

On the decay time of upconversion luminescence.

In this study, we systematically investigate the decay characteristics of upconversion luminescence (UCL) under anti-Stokes excitation through numerical simulations based on rate-equation models. We find that a UCL decay profile generally involves contributions from the sensitizer's excited-state lifetime, energy transfer and cross-relaxation processes. It should thus be regarded as the overall temporal response of the whole upconversion system to the excitation function rather than the intrinsic lifetime of the luminescence emitting state. Only under certain conditions, such as when the effective lifetime of the sensitizer's excited state is significantly shorter than that of the UCL emitting state and of the absence of cross-relaxation processes involving the emitting energy level, the UCL decay time approaches the intrinsic lifetime of the emitting state. Subsequently, Stokes excitation is generally preferred in order to accurately quantify the intrinsic lifetime of the emitting state. However, possible cross-relaxation between doped ions at high doping levels can complicate the decay characteristics of the luminescence and even make the Stokes-excitation approach fail. A strong cross-relaxation process can also account for the power dependence of the decay characteristics of UCL.

Colour-optimized quantum yields of Yb, Tm Co-doped upconversion nanocrystals.

We present here a systematic analysis of the influence of Tm3+ doping concentrations (x Tm ) on the excitation power (P)-dependent upconversion luminescence and -performance of hexagonal-phase NaYF4: 20% Yb3+, x Tm % Tm3+ upconversion nanoparticles (UCNPs) for x Tm of 0.2, 0.5, 0.8, 1.2, and 2.0, respectively. Our results reveal the influence of these different Tm3+ doping concentrations with respect to optimized upconversion quantum yield (Φ UC ) values of the various Tm3+ upconversion emission bands, with the highest Φ UC values of the Tm3+ emission bands above 700 nm resulting for different x Tm values as the Tm3+ emission bands below 700 nm. This underlines the potential of Tm3+ dopant concentration for colour tuning. Special emphasis was dedicated to the spectroscopic parameters that can be linked to the (de)population pathways of the various Tm3+ energy levels, like the P- and x Tm -dependent slope factors and the intensity ratios of selected emission bands. The evaluation of all parameters indicates that not only energy transfer upconversion-, but also cross-relaxation processes between neighbouring Tm3+ ions play a vital role in the (de)population of the excited energy levels of Yb3+, Tm3+ codoped nanocrystals.

Sensitization of upconverting nanoparticles with a NIR-emissive cyanine dye using a micellar encapsulation approach.

Photon upconversion nanomaterials have a wide range of applications, including biosensing and deep-tissue imaging. Their typically very weak and narrow absorption bands together with their size dependent luminescence efficiency can limit their application potential. This has been addressed by increasingly sophisticated core-shell particle architectures including the sensitization with organic dyes that strongly absorb in the near infrared (NIR). In this work, we present a simple water-dispersible micellar system that features energy transfer from the novel NIR excitable dye, 1859 SL with a high molar absorption coefficient and a moderate fluorescence quantum yield to oleate-capped NaYF4:20%Yb(III), 2%Er(III) upconversion nanoparticles (UCNP) upon 808 nm excitation. The micelles were formed using the surfactants Pluronic F-127 and Tween 80 to produce a hydrophilic dye-UCNP system. Successful energy transfer from the dye to the UCNP could be confirmed by emission measurements that revealed the occurrence of upconversion emission upon excitation at 808 nm and an enhancement of the green Er(III) emission compared to direct Er(III) excitation at 808 nm.

Absolute upconversion quantum yields of blue-emitting LiYF4:Yb3+,Tm3+ upconverting nanoparticles.

The upconversion quantum yield (ΦUC) is an essential parameter for the characterization of the optical performance of lanthanoid-doped upconverting nanoparticles (UCNPs). Despite its nonlinear dependence on excitation power density (Pexc), it is typically reported only as a single number. Here, we present the first measurement of absolute upconversion quantum yields of the individual emission bands of blue light-emitting LiYF4:Yb3+,Tm3+ UCNPs in toluene. Reporting the quantum yields for the individual emission bands is required for assessing the usability of UCNPs in various applications that require upconverted light of different wavelengths, such as bioimaging, photocatalysis and phototherapy. Here, the reliability of the ΦUC measurements is demonstrated by studying the same batch of UCNPs in three different research groups. The results show that whereas the total upconversion quantum yield of these UCNPs is quite high-typically 0.02 at a power density of 5 W cm-2-most of the upconverted photon flux is emitted in the 794 nm upconversion band, while the blue emission band at 480 nm is very weak, with a much lower quantum yield of ∼6 × 10-5 at 5 W cm-2. Overall, although the total upconversion quantum yield of LiYF4:Yb3+,Tm3+ UCNPs seems satisfying, notably for NIR bioimaging, blue-light demanding phototherapy applications will require better-performing UCNPs with higher blue light upconversion quantum yields.

NaYF4 :Yb,Er/NaYF4 Core/Shell Nanocrystals with High Upconversion Luminescence Quantum Yield.

Upconversion core/shell nanocrystals with different mean sizes ranging from 15 to 45 nm were prepared via a modified synthesis procedure based on anhydrous rare-earth acetates. All particles consist of a core of NaYF4 :Yb,Er, doped with 18 % Yb3+ and 2 % Er3+ , and an inert shell of NaYF4 , with the shell thickness being equal to the radius of the core particle. Absolute measurements of the photoluminescence quantum yield at a series of different excitation power densities show that the quantum yield of 45 nm core/shell particles is already very close to the quantum yield of microcrystalline upconversion phosphor powder. Smaller core/shell particles prepared by the same method show only a moderate decrease in quantum yield. The quantum yield of 15 nm core/shell particles, for instance, is reduced by a factor of three compared to the bulk upconversion phosphor at high power densities (100 W cm-2 ) and by approximately a factor of 10 at low power densities (1 W cm-2 ).

Quantum Yields, Surface Quenching, and Passivation Efficiency for Ultrasmall Core/Shell Upconverting Nanoparticles.

We synthesized and characterized a set of ultrasmall hexagonal-phase NaGdF4: 20% Yb3+, 2% Er3+ upconversion nanoparticles with core diameters of 3.7 ± 0.5 nm. In order to assess passivation effects and the influence of possible core-shell intermixing and to identify optimum particle structures for combined imaging in the visible and near-infrared (vis-NIR: 410-850 nm) and short-wave infrared (SWIR: 1520 nm), NaYF4 shells of varying thicknesses (monolayer to 10 nm) were introduced and the influence of this parameter on the upconversion and downshifting photoluminescence of these particles was studied at different excitation power densities. This included excitation power-dependent emission spectra, slope factors, quantum yields, and excited state decay kinetics. These measurements revealed enhancement factors of the upconversion quantum yield of >10 000 in the low power region and an excitation power density-independent quantum yield of the downshifted emission at 1520 nm between 0.1 and 14%. The optimized shell thickness for combined vis and SWIR imaging was identified as 5 nm. Moreover, lifetimes and quantum yields can be continuously tuned by shell thickness which can be exploited for lifetime multiplexing and encoding. The fact that we did not observe a saturation of the upconversion quantum yield or the excited state decay kinetics with increasing shell thickness is ascribed to a strong intermixing of the active core with the inert shell during the shelling procedure. This indicates the potential of spectroscopic tools to detect cation intermixing.

A protected excitation-energy reservoir for efficient upconversion luminescence.

Lanthanide-doped upconversion nanoparticles (UCNPs) are of great interest for biomedical applications. Currently, the applicability of UCNP bionanotechnology is hampered by the generally low luminescence intensity of UCNPs and inefficient energy transfer from UCNPs to surface-bound chromophores used e.g. for photodynamic therapy or analyte sensing. In this work, we address the low-efficiency issue by developing versatile core-shell nanostructures, where high-concentration sensitizers and activators are confined in the core and shell region of representative hexagonal NaYF4:Yb,Er UCNPs. After doping concentration optimization, the sensitizer-rich core is able to harvest/accumulate more excitation energy and generate almost one order of magnitude higher luminescence intensity than conventional homogeneously doped nanostructures. At the same time, the activator ions located in the shell enable a ∼6 times more efficient resonant energy transfer from UCNPs to surface-bound acceptor dye molecules due to the short distance between donor-acceptor pairs. Our work provides new insights into the rational design of UCNPs and will greatly increase the general applicability of upconversion nanotechnologies.

Shaping Luminescent Properties of Yb3+ and Ho3+ Co-Doped Upconverting Core-Shell ß-NaYF4 Nanoparticles by Dopant Distribution and Spacing.

At the core of luminescence color and lifetime tuning of rare earth doped upconverting nanoparticles (UCNPs), is the understanding of the impact of the particle architecture for commonly used sensitizer (S) and activator (A) ions. In this respect, a series of core@shell NaYF4 UCNPs doped with Yb3+ and Ho3+ ions are presented here, where the same dopant concentrations are distributed in different particle architectures following the scheme: YbHo core and YbHo@…, …@YbHo, Yb@Ho, Ho@Yb, YbHo@Yb, and Yb@YbHo core-shell NPs. As revealed by quantitative steady-state and time-resolved luminescence studies, the relative spatial distribution of the A and S ions in the UCNPs and their protection from surface quenching has a critical impact on their luminescence characteristics. Although the increased amount of Yb3+ ions boosts UCNP performance by amplifying the absorption, the Yb3+ ions can also efficiently dissipate the energy stored in the material through energy migration to the surface, thereby reducing the overall energy transfer efficiency to the activator ions. The results provide yet another proof that UC phosphor chemistry combined with materials engineering through intentional core@shell structures may help to fine-tune the luminescence features of UCNPs for their specific future applications in biosensing, bioimaging, photovoltaics, and display technologies.

Beam-profile-compensated quantum yield measurements of upconverting nanoparticles.

The quantum yield is a critically important parameter in the development of lanthanide-based upconverting nanoparticles (UCNPs) for use as novel contrast agents in biological imaging and optical reporters in assays. The present work focuses on the influence of the beam profile in measuring the quantum yield (ϕ) of nonscattering dispersions of nonlinear upconverting probes, by establishing a relation between ϕ and excitation light power density from a rate equation analysis. A resulting 60% correction in the measured ϕ due to the beam profile utilized for excitation underlines the significance of the beam profile in such measurements, and its impact when comparing results from different setups and groups across the world.