Photostability Of Dyes Research Articles

Coumarin-BODIPY hybrids by heteroatom linkage: versatile, tunable and photostable dye lasers for UV irradiation.

Linking amino and hydroxycoumarins to BODIPYs through the amino or hydroxyl group lets the easy construction of unprecedented photostable coumarin-BODIPY hybrids with broadened and enhanced absorption in the UV spectral region, and outstanding wavelength-tunable laser action within the green-to-red spectral region (∼520-680 nm). These laser dyes allow the generation of a valuable tunable UV (∼260-350 nm) laser source by frequency doubling, which is essential to study accurately the photochemistry of biological molecules under solar irradiation. The tunability is achieved by selecting the substitution pattern of the hybrid. Key factors are the linking heteroatom (nitrogen vs. oxygen), the number of coumarin units joined to the BODIPY framework and the involved linking positions.

Functionalization of the meso‐Phenyl Ring of Rhodamine Dyes Through SNAr with Sulfur Nucleophiles: Synthesis, Biophysical Characterizations, and Comprehensive NMR Analysis

AbstractAromatic nucleophilic substitution (SNAr) of fluorine in 9‐(3′‐carboxy‐4′,5′,6′,7′‐tetrafluorophenyl) groups of xanthene dyes constitutes a powerful tool in dye design. Thiols and amines regioselectively replace F‐6′. This approach enables additional hydrophilic residues or functional groups required for bioconjugation to be introduced. By using this methodology, a “bright” and photostable dye for two‐color superresolution microscopy was synthesized (with absorption and emission maxima at 604 and 627 nm, respectively). In the case of red‐emitting rhodamine dyes with 3′‐carboxy‐4′,5′,7′‐trifluorophenyl residues, two‐dimensional NMR techniques and a chemical transformation were used to prove the precise position of the additional substituent – a carboxylic acid group linked through the S‐atom at C‐6′. Furthermore, simple 1H NMR spectra reliably permit the position of the additional carboxy substituent in the 3′‐carboxyphenyl ring (at C‐5′ or C‐6′) to be established. Information on the exact position of this substituent is significant for the design of molecular probes and for the prediction of the properties of their bioconjugates.

Multifunctional liquid crystal nanoparticles for intracellular fluorescent imaging and drug delivery.

A continuing goal of nanoparticle (NP)-mediated drug delivery (NMDD) is the simultaneous improvement of drug efficacy coupled with tracking of the intracellular fate of the nanoparticle delivery vehicle and its drug cargo. Here, we present a robust multifunctional liquid crystal NP (LCNP)-based delivery system that affords facile intracellular fate tracking coupled with the efficient delivery and modulation of the anticancer therapeutic doxorubicin (Dox), employed here as a model drug cargo. The LCNPs consist of (1) a liquid crystal cross-linking agent, (2) a homologue of the organic chromophore perylene, and (3) a polymerizable surfactant containing a carboxylate headgroup. The NP core provides an environment to both incorporate fluorescent dye for spectrally tuned particle tracking and encapsulation of amphiphilic and/or hydrophobic agents for intracellular delivery. The carboxylate head groups enable conjugation to biologicals to facilitate the cellular uptake of the particles. Upon functionalization of the NPs with transferrin, we show the ability to differentially label the recycling endocytic pathway in HEK 293T/17 cells in a time-resolved manner with minimal cytotoxicity and with superior dye photostability compared to traditional organic fluorophores. Further, when passively loaded with Dox, the NPs mediate the rapid uptake and subsequent sustained release of Dox from within endocytic vesicles. We demonstrate the ability of the LCNPs to simultaneously serve as both an efficient delivery vehicle for Dox as well as a modulator of the drug's cytotoxicity. Specifically, the delivery of Dox as a LCNP conjugate results in a ∼40-fold improvement in its IC50 compared to free Dox in solution. Cumulatively, our results demonstrate the utility of the LCNPs as an effective nanomaterial for simultaneous cellular imaging, tracking, and delivery of drug cargos.

Multicolor fluorescence nanoscopy by photobleaching: concept, verification, and its application to resolve selective storage of proteins in platelets.

Fluorescence nanoscopy provides means to discern the finer details of protein localization and interaction in cells by offering an order of magnitude higher resolution than conventional optical imaging techniques. However, these super resolution techniques put higher demands on the optical system and the fluorescent probes, making multicolor fluorescence nanoscopy a challenging task. Here we present a new and simple procedure, which exploits the photostability and excitation spectra of dyes to increase the number of simultaneous recordable targets in STED nanoscopy. We use this procedure to demonstrate four-color STED imaging of platelets with ≤40 nm resolution and low crosstalk. Platelets can selectively store, sequester, and release a multitude of different proteins, in a manner specific for different physiological and disease states. By applying multicolor nanoscopy to study platelets, we can achieve spatial mapping of the protein organization with a high resolution for multiple proteins at the same time and in the same cell. This provides a means to identify specific platelet activation states for diagnostic purposes and to understand the underlying protein storage and release mechanisms. We studied the organization of the pro- and antiangiogenic proteins VEGF and PF-4, together with fibrinogen and filamentous actin, and found distinct features in their respective protein localization. Further, colocalization analysis revealed only minor overlap between the proteins VEGF and PF-4 indicating that they have separate storage and release mechanisms, corresponding well with their opposite roles as pro- and antiangiogenic proteins, respectively.

Scaffolding along nucleic acid duplexes using 2'-amino-locked nucleic acids.

CONSPECTUS: Incorporation of chemically modified nucleotide scaffolds into nucleic acids to form assemblies rich in function is an innovative area with great promise for nanotechnology and biomedical and material science applications. The intrinsic biorecognition potential of nucleic acids combined with advanced properties of the locked nucleic acids (LNAs) provide opportunities to develop new nanomaterials and devices like sensors, aptamers, and machines. In this Account, we describe recent research on preparation and investigation of the properties of LNA/DNA hybrids containing functionalized 2'-amino-LNA nucleotides. By application of different chemical reactions, modification of 2'-amino-LNA scaffolds can be efficiently performed in high yields and with various tags, postsynthetically or during the automated oligonucleotide synthesis. The choice of a synthetic method for scaffolding along 2'-amino-LNA mainly depends on the chemical nature of the modification, its price, its availability, and applications of the product. One of the most useful applications of the product LNA/DNA scaffolds containing 2'-amino-LNA is to detect complementary DNA and RNA targets. Examples of these applications include sensing of clinically important single-nucleotide polymorphisms (SNPs) and imaging of nucleic acids in vitro, in cell culture, and in vivo. According to our studies, 2'-amino-LNA scaffolds are efficient within diagnostic probes for DNA and RNA targets and as therapeutics, whereas both 2'-amino- and isomeric 2'-α-l-amino-LNA scaffolds have promising properties for stabilization and detection of DNA nanostructures. Attachment of fluorescent groups to the 2'-amino group results in very high fluorescent quantum yields of the duplexes and remarkable sensitivity of the fluorescence signal to target binding. Notably, fluorescent LNA/DNA probes bind nucleic acid targets with advantages of high affinity and specificity. Thus, molecular motion of nanodevices and programmable self-assembly of chemically modified LNA/DNA nanomaterials can be followed by bright fluorescence signaling from the functionalized LNA units. Another appealing aspect of the amino-LNA scaffolds is specific targeting of nucleic acids and proteins for therapeutic applications. 2'-Amino-LNA/DNA conjugates containing peptide and polyaromatic hydrocarbon (PAH) groups are promising in this context as well as for advanced imaging and diagnostic purposes in vivo. For imaging applications, photostability of fluorescence dyes is of crucial importance. Chemically stable and photostable fluorescent PAH molecules attached to 2'-amino functionality of the 2'-amino-LNA are potent for in vitro and in vivo imaging of DNA and RNA targets. We believe that rational evolution of the biopolymers of Nature may solve the major challenges of the future material science and biomedicine. However, this requires strong scientific progress and efficient interdisciplinary research. Examples of this Account demonstrate that among other synthetic biopolymers, synthetic nucleic acids containing functionalized 2'-amino-LNA scaffolds offer great opportunities for material science, diagnostics, and medicine of the future.

Transmission booster from SiO2 incorporated TiO2 crystallites: Enhanced conversion efficiency in dye-sensitized solar cells

Dye-sensitized solar cell (DSSC) is a promising solution to energy depletion, environmental pollution, and ecological destruction. However, the further elevation of power conversion efficiency is limited by low electron density on conduction band (CB) of TiO2 crystallite, resulting from the low dye-excitation efficiency, recombination of electrons with ions in liquid electrolyte as well as photodegradation of dye molecules. With an aim of enhancing the power conversion efficiency, here we designed SiO2 incorporated TiO2 crystallite photoanode to elevate dye excitation, to suppress the recombination of electron-electrolyte, and to enhance the dye photostability. We found that the highest conversion occurred at 1 wt% SiO2 incorporated TiO2 crystallite-based DSSC, giving an impressive power conversion efficiency of 9.98% (measured under standard AM1.5 test conditions) in comparison with 6.13% from pure TiO2 photoanode-based DSSC. This strategy provides new opportunities for the fabrication of highly efficient DSSCs.

Transmission enhanced photoanodes for efficient dye-sensitized solar cells

The power conversion efficiency elevation of dye-sensitized solar cell (DSSC) is limited by the incomplete dye excitation, recombination of photogenerated electrons with ions in liquid electrolyte, low electron density in conduction band (CB) of TiO2 nanoscrystalline as well as photodegradation of dye molecules. With an aim of enhancing the power conversion efficiency, here we describe an employment of SiO2 incorporated TiO2 to enhance transmission, to suppress the recombination of electron-electrolyte, to elevate the dye loading and to increase the dye photostability. We find that the highest conversion occurs at 7 wt% SiO2 in SiO2 incorporated TiO2 nanocrystallines is 9.13% (measured under standard AM1.5 test conditions) in comparison with 6.26% from pure TiO2 photoanode, giving an efficiency enhancement by 45.8%. This strategy provides new opportunities for the fabrication of highly efficient DSSCs, and the efficiency record is expected to be surpassed by scalable technique and components.

Role of Dead Box Helicases in HIV-1 Rev Function: a Single-Molecule Approach

The HIV-1 Rev (Regulator of Expression of Virion) protein activates nuclear export of unspliced and partially spliced viral mRNAs, which encode the viral genome and the genes encoding viral structural proteins. Rev interacts with a highly conserved region, the Rev Response Element (RRE), located within the viral mRNA. Initially, a single Rev monomer binds to stem loop IIB of the RRE, whereupon additional Rev monomers are recruited to the RRE through a combination of RNA-protein and protein-protein interactions, resulting in the formation of a functional nuclear export complex. In addition, several cellular host proteins, such as the DEAD box helicases DDX1 and DDX21 are known to be required for efficient Rev function in vivo, although their precise role is unknown. In this study, a variety of single-molecule fluorescence spectroscopic methods were used to dissect the role of DDX1 and DDX21 during assembly of Rev-RRE complexes. To facilitate these studies, the large DDX1 and DDX21 proteins were enzymatically labeled with bright and photostable dyes, while Rev was labeled at a single cysteine by maleimide chemistry. Single-color TIRF measurements with labeled Rev were used to monitor individual Rev monomer binding steps during oligomeric Rev-RRE assembly. Moreover, single-color experiments with labeled DDX1 or DDX21 reveal that each helicase is capable of binding directly to the RRE. Two-color colocalization and FRET measurements were used to monitor the simultaneous binding of both Rev and DDX1 to the RRE, revealing the temporal correlation between individual protein binding steps. Together, these studies are revealing how DEAD box helicases are able to promote the oligomeric assembly of Rev on the RRE, thereby acting as cellular cofactors of HIV-1. Supported by NIH P50 grant GM082545.

Improved Super-Resolution Imaging in Heavy Water

Advanced fluorescence microscopy techniques including single-molecule and super-resolution imaging require bright and photostable dyes that can be selectively targeted to biomolecules. There is therefore an ongoing interest in the development of improved chromophores for biology, especially ones that absorb and emit in the near-infrared. Single fluorophore brightness is a key parameter in localization-based super-resolution imaging techniques such as (F)PALM, (d)STORM, or GSDIM: the brighter the emitter, the more precisely it can be localized and the better the resolution is in the final image. Nonetheless, with few exceptions, most commercially available fluorophores excitable beyond 630 nm have poor fluorescence quantum yields with values around 0.3 or lower limiting their brightness.We introduce a simple, cost-effective, and biocompatible method to enhance the fluorescence quantum yield of several commercially available oxazine (ATTO655, ATTO680, ATTO700) and cyanine (Cy5, Alexa Fluor 647, Cy5.5, Alexa Fluor 700, Cy7) fluorophores [1, 2]. We demonstrate that for all oxazines and one cyanine, the fluorescence quantum yield more than doubles in heavy water (D2O) compared to water (H2O). This increase leads to the detection of twice as many photons per molecule and to a corresponding improvement in the mean localization precision, which enables higher resolution in biological subdiffraction imaging. We demonstrate that super-resolution imaging in D2O can be used to resolve diffraction-limited subcellular structures such as filopodia or microtubules and by performing quantitative morphological analysis of ligand-induced clustering of arrestin proteins upon G protein-coupled receptor stimulation [3].[1] S. F. Lee, Q. Verolet, A. Furstenberg, Angew. Chem. Int. Ed. 2013, 52, 8948.[2] K. Klehs, C. Spahn, U. Endesfelder, S. F. Lee, A. Furstenberg, M. Heilemann, submitted.[3] Z. Truan, L. Tarancon Diez, C. Bonsch, S. Malkusch, U. Endesfelder, M. Munteanu, O. Hartley, M. Heilemann, A. Furstenberg, J. Struct. Biol., in press.

Super Resolution Mapping of Adhesion Molecules in Confined Cellular Environments using Monomeric Streptavidin Ligands

High resolution tracking of individual cell surface proteins is limited by several factors, including probe size that can impair protein trafficking and restrict their access to crowded cellular environments, ligand multivalency that induces protein cross-linking, and potential loss of function of recombinant proteins fused to large tags. To overcome these issues, we developed a labeling method that uses monomeric streptavidin (mStrav) conjugated to highly photo-robust organic Atto dyes. mStrav is a 3-nm-molecule, stable and easy to produce, that binds biotinylated proteins with high affinity (Lim et al., Biochemistry 2011). Atto-conjugated mStrav is then used as a probe to label cells co-transfected with a membrane molecule of interest carrying a 15 amino acid extra-cellular acceptor peptide (AP), and the biotin ligase BirA that covalently adds biotin to the tag during protein maturation. We used this technique to study membrane diffusion and nanoscale organization of the synaptic adhesion molecules neurexin-1β and neuroligin-1 in hippocampal neurons and β3-integrins in fibroblasts. We show that the monovalency of mStrav prevents protein cross-linking, allowing more accurate diffusion measurements. The small sizes of the AP tag (<1nm) and mStrav (∼3nm) do not perturb protein function and allow access to single molecule dynamics and high resolution mapping in confined regions such as synapses or cell-matrix contacts. We also used mStrav along with nanobodies (∼3nm) for dual-color single molecule tracking. Moreover, the binding of mStrav can be reversed with excess biotin, a property that can be used to study protein endocytosis and recycling. Finally, the photostability of Atto dyes conjugated to mStrav allows for long trajectory recordings and live STED imaging. Taken together, monomeric streptavidin is a simple and versatile tool that can be used to track an arbitrary membrane protein in living cells.

Diffusion Mapping in Living Cells using Camera-Based Correlation Spectroscopy and Phasor Analysis

Diffusion Mapping in Living Cells using Camera-Based Correlation Spectroscopy and Phasor Analysis

Photostability of 4,4′‐Dihydroxythioindigo, a Mimetic of Indigo

The photochemical properties of indigo, a widely used industrial dye, has attracted both experimentalists and theoreticians from the beginning. Especially the high photostability of indigo has been the subject of intensive research. Recently, it was proposed that after photoexcitation an intramolecular proton transfer followed by a nonradiative relaxation to the ground state promote photostability. In indigo the hydrogen bond and the proton transfer occur between the opposing hemiindigo parts. Here, we provide experimental and theoretical evidence that a hydrogen transfer within one hemiindigo or hemithioindigo part is sufficient to attain photostability. This concept can serve as an interesting strategy towards new photostable dyes for the visible part of the spectrum.

Synthesis and performance of coumarin containing polymeric fluorescent dyes

Purpose – The purpose of this paper is to modify the photostability of organic fluorescent dyes and to develop and evaluate a route of their derived polymeric fluorescent dyes. Design/methodology/approach – A series of novel polymeric fluorescent dyes were prepared using 3-(2-benzimidazolyl)-7-(diethylamino)-coumarin (disperse yellow 8GFF, abbreviated as 8GFF) and polyethylene glycol (PEG) as raw materials. First, an intermediate of carboxyl functioned 8GFF was prepared by condensation reaction of 8GFF and pyromellitic dianhydride; and then the polymeric fluorescent dyes were synthesized by reacting PEG with the condensate intermediate. Findings – FTIR, UV-vis, and PL showed that the polymeric fluorescent dyes had been successfully synthesised, and the photostability of organic fluorescent dyes was improved significantly. Research limitations/implications – In the present work, the polymeric fluorescent dyes had been successfully synthesized, and the photostability of organic fluorescent dyes was improved significantly. This methodology can be employed to prepare novel polymeric fluorescent materials and to improve the photostability of organic dyes for various applications. Practical implications – Polycondensable dyes offer a new area of material science which can be widely used in coloured polymer materials production. Originality/value – The method developed in the study reported in this paper provided a new strategy to develop new types of polycondensable dyes materials with fluorescent property.

Relating Electron Donor and Carboxylic Acid Anchoring Substitution Effects in Azo Dyes to Dye-Sensitized Solar Cell Performance

The relationship between the molecular structures of a series of azo dyes and their operational performance when applied to dye-sensitized solar cells (DSSCs) is probed via experimental and computational analysis. Seven azo dyes, with three different donating groups (dimethylamino, diethylamino, and dipropylamino) and carboxylic acid anchoring positions (ortho-, meta-, and para-substituted phenyl rings) are studied. Single-crystal X-ray diffraction is employed in order to analyze the effects of conformation and quantify the contribution of quinoidal resonance forms to the intramolecular charge transfer (ICT), which controls their intrinsic photovoltaic potential from an electronic standpoint. Harmonic oscillator stabilization energy (HOSE) calculations indicate that the para- and ortho-azo dyes exhibit potential for DSSC application. However, from a geometrical standpoint, the crystal structure data, proton nuclear magnetic resonance spectroscopy (1H NMR), and density functional theory (DFT) all indicate that intramolecular hydrogen bonds form in ortho-dyes within both solid and solution states, impeding their intrinsic ICT-based photovoltaic potential, and offering insights into the photostability of azo dyes and the dye···TiO2 anchoring mechanism in DSSCs. Donor effects are manifested in the packing mode and molecular planarity revealed by X-ray crystallography and in the UV/vis absorption spectra. DFT and time-dependent density functional theory (TDDFT) were performed to understand the electronic and optical properties of these azo dyes; these calculations compare well with experimental findings. Operational tests of DSSCs, functionalized by these azo dyes, show that the carboxylic acid anchoring position plays a crucial role in DSSC performance, while donating groups offer a much less obvious effect on the overall DSSC device efficiency.

Micelles assembled with carbocyanine dyes for theranostic near-infrared fluorescent cancer imaging and photothermal therapy

It is an emerging focus to explore a theranostic nanocarrier for simultaneous cancer imaging and therapy. Herein, we demonstrate a theranostic micelle system for cancer near infrared fluorescent (NIRF) imaging with enhanced signal to noise ratio and superior photothermal therapy. The copolymers consisting of monomethoxy poly(ethylene glycol) and alkylamine-grafted poly(l-aspartic acid) are assembled with carbocyanine dyes into theranostic micelles, which exhibit small size, high loading capacity, good stability, sustained release behavior, and enhanced cellular uptake. The micelles achieve the preferable biodistribution and long-term retention of carbocyanine dyes at tumor, which result in enhanced NIRF imaging by generating stable retention of NIRF signals at both hypervascular and hypovascular tumors during a long-term imaging period of up to 8 day, accompanying with negligible noise at normal tissues. The photostability of carbocyanine dye (Cypate) plays an important role for long-term cancer imaging with enhanced SNR. Moreover, the micelles exhibit severe photothermal damage on cancer cells via the destabilization of subcellular organelles upon photoirradiation, causing superior photothermal tumor regress. The micelles act as a powerful theranostic nanocarrier for simultaneous cancer imaging with high contrast and superior photothermal therapy.

The photoinduced transformation of fluorescent DNA base analogue tC triggers DNA melting

While fluorescent analogues of the canonical nucleobases have proven to be highly valuable in a large number of applications, up until today, fluorescent DNA base analogues remain virtually inapplicable for single-molecule fluorescence experiments which require extremely bright and photostable dyes. Insight into the photodegradation processes of these fluorophores is thus a key step in the continuous development towards dyes with improved performances. Here, we show that the commercially available fluorescent nucleobase analogue tC under intense long-term illumination and in the presence of O2 is degraded to form a single photoreaction product which we suggest to be the sulfoxide form of tC. The photoproduct is characterized by a blue-shifted absorption and a less intense fluorescence compared to that of tC. Interestingly, when tC is positioned inside double-stranded DNA this photodriven conversion of tC to its photoproduct greatly reduces the duplex stability of the overall double helix in which the probe is positioned. Since tC can be excited selectively at 400 nm, well outside the absorption band of the natural DNA bases, this observation points towards the application of tC as a general light-triggered switch of DNA duplex stability.

Plasmonic-based instrument response function for time-resolved fluorescence: toward proper lifetime analysis

In this report, we investigated the so-called plasmonic platforms prepared to target ultra-short fluorescence and accurate instrumental response function in a time-domain spectroscopy and microscopy. The interaction of metallic nanoparticles with nearby fluorophores results in the increase of the dye fluorescence quantum yield, photostability and decrease of the lifetime parameter. The mentioned properties of platforms were applied to achieve a picosecond fluorescence lifetime (21 ps) of erythrosin B, used later as a better choice for deconvolution of fluorescence decays measured with “color” sensitive photo-detectors. The ultra-short fluorescence standard based on combination of thin layers of silver film, silver colloidal nanoparticles (about 60 nm in diameter), and top layer of erythrosin B embedded in 0.2 % poly(vinyl) alcohol. The response functions were monitored on two photo-detectors; microchannel plate photomultiplier and single photon avalanche photodiode as a Rayleigh scattering and ultra-short fluorescence. We demonstrated that use of the plasmonic base fluorescence standard as an instrumental response function results in the absence of systematic error in lifetime measurements and analysis.

Fluorinated Boron‐Dipyrromethene (BODIPY) Dyes: Bright and Versatile Probes for Surface Analysis

A family of bright boron-dipyrromethene-type fluorophores with a high number of fluorine atoms (F-BODIPYs) has been developed and characterized by X-ray crystallography and optical spectroscopy. The introduction of 3,5-bis(trifluoromethyl)phenyl and pentafluorophenyl moieties significantly enhances the photostability of such dyes, yielding for instance photostable near-infrared (NIR) fluorophores that show emission maxima>750 nm, when the BODIPY’s π system is extended with two (dimethylamino)styryl and (dimethylamino)naphthastyryl moieties, or green-emitting BODIPYs with fluorescence quantum yields of unity. When equipped with a suitable group that selectively reacts for instance with amines, F-BODIPYs can be used as potent dual labels for the quantification of primary amino groups on surfaces by X-ray photoelectron spectroscopy (XPS) and fluorescence, two powerful yet complementary tools for the analysis of organic surface functional groups. The advantage of reactive F-BODIPYs is that they allow a fast and non-destructive mapping of the labelled supports with conventional fluorescence scanners and a subsequent quantification of selected areas of the same sample by the potentially traceable XPS technique. The performance is exemplarily shown here for the assessment of the amino group density on SiO2 supports, one of the most common reactive silica supports, in particular, for standard microarray applications.

Synthesis and evaluation of cyanine–styryl dyes with enhanced photostability for fluorescent DNA staining

The photostability of cyanine-styryl dyes of the indole-quinolinium type can be significantly improved by structural variations while the excellent optical properties including the bright fluorescence in the presence of DNA can be maintained or even improved, too.

Full-color tunable organic nanoparticles with FRET-assisted enhanced two-photon excited fluorescence for bio-imaging.

Two-photon excited fluorescence (TPEF) bio-probes are important for tissue imaging. Here, we demonstrated a strategy for highly emissive, photostable, low cytotoxicity and full-color tunable TPEF organic nanoparticles (ONPs) comprising of commercially available dyes doped in a TPEF host matrix. Because of efficient Förster resonance energy transfer (FRET), near-IR laser pumping light can be strongly absorbed by host matrix molecules via a two-photon absorption (TPA) process and transferred to dopant dyes, which then emit visible photons in different colors from blue, green, and yellow to red with high quantum yields. Our strategy significantly enhanced the photo-stability and TPEF of dopant dyes in ONPs, avoiding the need to resort to specially engineered expensive dyes.