Solvatochromic Probe Research Articles

Mapping membrane biophysical nano-environments

The mammalian plasma membrane is known to contain domains with varying lipid composition and biophysical properties. However, studying these membrane lipid domains presents challenges due to their predicted morphological similarity to the bulk membrane and their scale being below the classical resolution limit of optical microscopy. To address this, we combine the solvatochromic probe di-4-ANEPPDHQ, which reports on its biophysical environment through changes in its fluorescence emission, with spectrally resolved single-molecule localisation microscopy. The resulting data comprises nanometre-precision localisation coordinates and a generalised polarisation value related to the probe’s environment – a marked point pattern. We introduce quantification algorithms based on topological data analysis (PLASMA) to detect and map nano-domains in this marked data, demonstrating their effectiveness in both artificial membranes and live cells. By leveraging environmentally sensitive fluorophores, multi-modal single molecule localisation microscopy, and advanced analysis methods, we achieve nanometre scale mapping of membrane properties and assess changes in response to external perturbation with methyl-β-cyclodextrin. This integrated methodology represents an integrated toolset for investigating marked point pattern data at nanometre spatial scales.

Multi-focal lipid membrane characterization by combination of DAS-deconvolution and anisotropy: Novel insight of fluorescence analysis

Three analogue solvatochromic probes, Laurdan, Prodan, and Acdan, are extensively used in the study of biological sciences. Their locations in lipid membranes vary greatly in depth, and their fluorescence responds to their surrounding environment based on their corresponding locations in the membrane. Utilizing the fluorescence lifetimes (τ) and emission peaks (λ) acquired from time-resolved emission spectrum, one can effectively determine the local lipid environment using the analytical approach refer to as τ and λ plots. Herein, τ and λ plot was created using the aforementioned probes to expand the analytical field according to their location. Furthermore, the solvent modeling method in τ and λ plot was upgraded to artificially emulate the complex environment in lipid membranes by utilizing liquid paraffin and glycerol to assess the contribution of viscosity to each fluorescence distribution. According to the results from a series of solvent mixtures, the effect of solvent viscosity on lifetime values was confirmed in the short lifetime region (∼ 3 ns). However, it was impossible to emulate the long lifetime values (4 ns ∼) observed in lipid membranes containing 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in the range of viscosity applied in this study. From the insight of the limiting anisotropy (r∞), the τ and λ plot was divided into a solvent-like region with isotropic environment (r∞ < 0.15) and a highly ordered region enough to define as anisotropic environment (0.15 <r∞) at τ: 4 ns. Also, the membrane-specific distribution was illustrated as 4 ns <τ and λ < 460 nm from this work. Updated analytical model was created to visualize multiple fluorescence components of each probe in 6 types of lipid bilayers, confirming the different distributions between these probes. Our results well-illustrated the multiplicity of lipid environments modeled with solvent and ordered environments in each lipid bilayer systems.

Intramolecular Charge Transfer Inhibition Strategy toward a Desired Solvatochromic Fluorescent Platform: Visualization of Duple Organelles and Detection of Carbon Dioxide.

Solvatochromic fluorescent probes are crucial molecular tools to investigate and aggregate proteins' fold, visualize fine structures in biomembranes, and label different organelles in dual emission colors. However, solvatochromic fluorogens often displayed a weak emission at high polarity, hindering their bioimaging applications. To resolve this problem, herein, we propose an intramolecular charge transfer (ICT) inhibition strategy. The probe was designed with a single electronic donor and two acceptors in order to split and inhibit the ICT procedure. As a result, the probe displayed an intense emission at both low and high polarities and showed a large emission shift (84 nm) upon polarity change. Using the probe, we successfully imaged lipid droplets and the endoplasmic reticulum in different fluorescence colors. Moreover, the different degrees of lipid accumulation by oleic acid, stearic acid, and cholesterol (oleic acid > stearic acid > cholesterol) have been revealed. The lipid accumulation induced by the three lipids could be rapidly consumed under lipid-less conditions, and the lipids with stearic acid were the most difficult to be consumed. The biological results could facilitate the understanding and treatment of lipid accumulation and obesity. Furthermore, utilizing the polarity increase of diethylamine after the reaction with CO2, the ratiometric detection of CO2 has been achieved for the first time with the probe.

Integrating computational and experimental approaches in discovery and validation of MmpL3 pore domain inhibitors for specific labelling of Mycobacterium tuberculosis

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), continues to pose a significant global health threat. Identifying new druggable targets is crucial for the advancement of drug development. Equally critical is the development of precise methods for monitoring Mtb to effectively combat this disease. Addressing these needs, our study pinpointed the pore domain (PD) of MtbMmpL3 as a new binding site for virtual screening, which led to the discovery of the small molecule ZY27. To confirm the binding site and action mode of ZY27, we employed cosolvent molecular dynamics (CMD), steered molecular dynamics (SMD), and long timescale molecular dynamics (MD) simulations of 5 μs. These in silico studies verified that ZY27 binds to the PD of MtbMmpL3. In antimicrobial activity tests, ZY27 exhibited potent anti-Mtb activity and high selectivity among mycobacterial species. Whole-genome sequencing of spontaneous ZY27-resistant Mtb variants, complemented by acid-fast staining experiments, confirmed that ZY27 specifically targets MtbMmpL3. Utilizing the ligand-protein binding data, we designed and synthesized two solvatochromic fluorescent probes, 27FP1 and 27FP2, based on ZY27. Further investigations through flow cytometry and confocal microscopy confirmed that these probes specifically label Mtb cells via the MtbMmpL3 binding mechanism.

Naphthalimide Based Fluorophore for the Detection of Hazardous Volatile Organic Compounds (VOCs).

The naphthalimide molecule (NAP) was successfully synthesized and characterized by spectroscopy techniques. The NAP probe was exposed to a solvatochromic and aggregation-induced emission (AIE) probe using UV-visible and PL spectroscopy. In this case, the increased polarity of the solvent shows that it is red-shifted. The probe emission, a sky blue to yellow-green color at hexane to DCM, exhibited an excellent quantum yield. Meanwhile, the high-polar solvents of DMF and DMSO had poor quantum yields. This probe NAP showed the aggregation-induced emission property dramatically enhanced the emissions (at 540nm) from fw = 80-90%. NAP was conducted with two polar solvent vapors in hexane and chloroform to investigate VOCs in a further solid-state study. A real-life test paper kit NAP probe was prepared and investigated VOCs detection against hexane and chloroform. When exposed to hexane vapors, the NAP probe test kit showed sky blue emission under UV light, which returned to greenish emission upon exposure to chloroform. Therefore, the results show that this NAP probe can be used for gas leak detection applications.

Organelle-Targeted Laurdans Measure Heterogeneity in Subcellular Membranes and Their Responses to Saturated Lipid Stress.

Organelles feature characteristic lipid compositions that lead to differences in membrane properties. In cells, membrane ordering and fluidity are commonly measured using the solvatochromic dye Laurdan, whose fluorescence is sensitive to lipid packing. As a general lipophilic dye, Laurdan stains all hydrophobic environments in cells; therefore, it is challenging to characterize membrane properties in specific organelles or assess their responses to pharmacological treatments in intact cells. Here, we describe the synthesis and application of Laurdan-derived probes that read out the membrane packing of individual cellular organelles. The set of organelle-targeted Laurdans (OTL) localizes to the ER, mitochondria, lysosomes, and Golgi compartments with high specificity while retaining the spectral resolution needed to detect biological changes in membrane ordering. We show that ratiometric imaging with OTLs can resolve membrane heterogeneity within organelles as well as changes in lipid packing resulting from inhibition of trafficking or bioenergetic processes. We apply these probes to characterize organelle-specific responses to saturated lipid stress. While the ER and lysosomal membrane fluidity is sensitive to exogenous saturated fatty acids, that of mitochondrial membranes is protected. We then use differences in ER membrane fluidity to sort populations of cells based on their fatty acid diet, highlighting the ability of organelle-localized solvatochromic probes to distinguish between cells based on their metabolic state. These results expand the repertoire of targeted membrane probes and demonstrate their application in interrogating lipid dysregulation.

Photophysics in Biomembranes: Computational Insight into the Interaction between Lipid Bilayers and Chromophores.

ConspectusLight is ubiquitously available to probe the structure and dynamics of biomolecules and biological tissues. Generally, this cannot be done directly with visible light, because of the absence of absorption by those biomolecules. This problem can be overcome by incorporating organic molecules (chromophores) that show an optical response in the vicinity of those biomolecules. Since those optical properties are strongly dependent on the chromophore's environment, time-resolved spectroscopic studies can provide a wealth of information on biosystems at the molecular scale in a nondestructive way. In this work, we give an overview on the multiscale computational strategy developed by us in the last eight years and prove that theoretical studies and simulations are needed to explain, guide, and predict observations in fluorescence experiments. As we challenge the accepted views on existing probes, we discover unexplored abilities that can discriminate surrounding lipid bilayers and their temperature-dependent as well as solvent-dependent properties. We focus on three archetypal chromophores: diphenylhexatriene (DPH), Laurdan, and azobenzene. Our method shows that conformational changes should not be neglected for the prototype rod-shaped molecule DPH. They determine its position and orientation in a liquid-ordered (Lo) sphingomyelin/cholesterol (SM/Chol) bilayer and are responsible for a strong differentiation of its absorption spectra and fluorescence decay times in dioleoylphosphatidylcholine (DOPC) and dipalmitoylphosphatidylcholine (DPPC) membranes, which are at room temperature in liquid-disordered (Ld) and solid-gel (So) phases, respectively. Thanks to its pronounced first excited state dipole moment, Laurdan has long been known as a solvatochromic probe. Since this molecule has however two conformers, we prove that they exhibit different properties in different lipid membrane phases. We see that the two conformers are only blocked in one phase but not in another. Supported by fluorescence anisotropy decay simulations, Laurdan can therefore be regarded as a molecular rotor. Finally, the conformational versatility of azobenzene in saturated Ld lipid bilayers is simulated, along with its photoisomerization pathways. By means of nonadiabatic QM/MM surface hopping analyses (QM/MM-SH), a dual mechanism is found with a torsional mechanism and a slow conversion for trans-to-cis. For cis-to-trans, simulations show a much higher quantum yield and a so-called "pedal-like" mechanism. The differences are related to the different potential energy surfaces as well as the interactions with the surrounding alkyl chains. When tails of increased length are attached to this probe, cis is pushed toward the polar surface, while trans is pulled toward the center of the membrane.

Hydration- and Temperature-Dependent Fluorescence Spectra of Laurdan Conformers in a DPPC Membrane.

The widely used Laurdan probe has two conformers, resulting in different optical properties when embedded in a lipid bilayer membrane, as demonstrated by our previous simulations. Up to now, the two conformers' optical responses have, however, not been investigated when the temperature and the phase of the membrane change. Since Laurdan is known to be both a molecular rotor and a solvatochromic probe, it is subject to a profound interaction with both neighboring lipids and water molecules. In the current study, molecular dynamics simulations and hybrid Quantum Mechanics/Molecular Mechanics calculations are performed for a DPPC membrane at eight temperatures between 270K and 320K, while the position, orientation, fluorescence lifetime and fluorescence anisotropy of the embedded probes are monitored. The importance of both conformers is proven through a stringent comparison with experiments, which corroborates the theoretical findings. It is seen that for Conf-I, the excited state lifetime is longer than the relaxation of the environment, while for Conf-II, the surroundings are not yet adapted when the probe returns to the ground state. Throughout the temperature range, the lifetime and anisotropy decay curves can be used to identify the different membrane phases. The current work might, therefore, be of importance for biomedical studies on diseases, which are associated with cell membrane transformations.

60 Years of Betaine 30─From Solvatochromic Discovery to Future Frontiers.

Betaine-30 (B30) was reported by Karl Dimroth and Christian Reichardt et al. in 1963 as a solvatochromic probe that can be easily synthesized, shows good solubility, and remains stable in various organic solvents and solutions. Its strongly negatively solvatochromic behavior arises from differential solvation between its electronic ground and excited states, making it a valuable tool for assessing solvent polarity using the ET(30) polarity scale, also devised by Dimroth and Reichardt. In addition, advancements in femtosecond laser spectroscopy in the 1990s greatly improved the understanding of B30's relaxation dynamics following photoexcitation. In solvents capable of hydrogen bonding, such as alcohols, intermolecular hydrogen-bond rearrangement contributes to the multiple relaxation components observed. Since the 1990s, the applications of B30 have expanded beyond simple organic solvents to include complex solvent mixtures, such as electrolyte solutions for battery technologies and eutectic solvent mixtures. Given the growing importance of these complex solvent mixtures, B30 is becoming an increasingly valuable tool for studying previously unexplored solvation properties.

Deeper Insights into Mixed Crowding through Enzyme Activity, Dynamics, and Crowder Diffusion.

Given the fact that the cellular interior is crowded by many different kinds of macromolecules, it is important that in vitro studies be carried out in the presence of mixed crowder systems. In this regard, we have used binary crowders formed by the combination of some of the commonly used crowding agents, namely, Ficoll 70, Dextran 70, Dextran 40, and PEG 8000 (PEG 8), to study how these affect enzyme activity, dynamics, and crowder diffusion. The enzyme chosen is AK3L1, an isoform of adenylate kinase. To investigate its dynamics, we have carried out three single point mutations (A74C, A132C, and A209C) with the cysteine residues being labeled with a coumarin-based solvatochromic probe [CPM: (7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin)]. Both enzyme activity and dynamics decreased in the binary mixtures as compared with the sum of the individual crowders, suggesting a reduction in excluded volume (in the mixture). To gain deeper insights into the binary mixtures, fluorescence correlation spectroscopy studies were carried out using fluorescein isothiocyanate-labeled Dextran 70 and tetramethylrhodamine-labeled AK3L1 as the diffusion probes. Diffusion in binary mixtures was observed to be much more constrained (relative to the sum of the individual crowders) for the labeled enzyme as compared to the labeled crowder showing different environments being faced by the two species. This was further confirmed during imaging of the phase-separated droplets formed in the binary mixtures having PEG as one of the crowding agents. The interior of these droplets was found to be rich in crowders and densely packed, as shown by confocal and digital holographic microscopy images, with the enzymes predominantly residing outside these droplets, that is, in the relatively less crowded regions. Taken together, our data provide important insights into various aspects of the simplest form of mixed crowding, that is, composed of just two components, and also hint at the enhanced complexity that the cellular interior presents toward having a detailed and comprehensive understanding of the same.

Umbrella‐Shaped Amphiphiles: Internal Alkylation of an Aromatic Micelle and Its Impact on Cavity Features

AbstractTo develop new hybrid micelles with alkyl/polyaromatic core–shell structures, we synthesized umbrella‐shaped amphiphiles bearing a bent anthracene dimer with a linear alkyl chain (i.e., octyl and hexadecyl groups). The amphiphiles quantitatively assemble into spherical micelles (~2–3 nm in core diameter), possessing an alkylated cavity surrounded by a polyaromatic framework, in water. The alkylation significantly enhances the stability of the micellar structures against dilution (up to 9 μM) and heat (up to &gt;120 °C). The highly condensed hexadecyl core of the hybrid micelle, as indicated by solvatochromic guest probes, displays increased uptake ability toward large alkylated metallodyes. Interestingly, efficient uptake of aromatic macrocycles (i.e., [n]cycloparaphenylenes) by the present micelle provides pseudorotaxane‐shaped host–guest composites with high emissivity (ΦF=up to 35 %). Internal multi‐alkylation of an aromatic micelle can thus successfully enhance its assembly stability/guest uptake functions.

Umbrella-Shaped Amphiphiles: Internal Alkylation of an Aromatic Micelle and Its Impact on Cavity Features.

To develop new hybrid micelles with alkyl/polyaromatic core-shell structures, we synthesized umbrella-shaped amphiphiles bearing a bent anthracene dimer with a linear alkyl chain (i.e., octyl and hexadecyl groups). The amphiphiles quantitatively assemble into spherical micelles (~2-3 nm in core diameter), possessing an alkylated cavity surrounded by a polyaromatic framework, in water. The alkylation significantly enhances the stability of the micellar structures against dilution (up to 9 μM) and heat (up to >120 °C). The highly condensed hexadecyl core of the hybrid micelle, as indicated by solvatochromic guest probes, displays increased uptake ability toward large alkylated metallodyes. Interestingly, efficient uptake of aromatic macrocycles (i.e., [n]cycloparaphenylenes) by the present micelle provides pseudorotaxane-shaped host-guest composites with high emissivity (ΦF=up to 35 %). Internal multi-alkylation of an aromatic micelle can thus successfully enhance its assembly stability/guest uptake functions.

A low-molecular-weight, long-wavelength emissive Coumarin: Green synthesis, spectral properties, fluorescence enhancement and solvatochromism application

In this study, a novel low-molecular-weight coumarin fluorescent dye, 3,4-dicyano-7-diethylaminocoumarin, was designed. This dye features a donor-π-bridge-acceptor (D-π-A) conjugated structure, which confers superior fluorescence properties.The dye was synthesized using a simple, efficient, and environmentally friendly two-step process, facilitating the direct conversion of the key intermediate into the final product via simultaneous addition and elimination reactions. A comprehensive structural characterization of the dye was undertaken, and its UV absorption and fluorescence emission properties were extensively analyzed using spectroscopic techniques, with structural explanations of the fluorescence activities provided by the Density Functional Theory (DFT) method.Experimental results revealed that this coumarin dye emits strong fluorescence, characterized by a long wavelength, a large Stokes shift, and high fluorescence intensity. It exhibits excellent fluorescent performance in various solvents. Despite experiencing fluorescence quenching in its solid state, the formation of an inclusion complex with β-cyclodextrin restores strong red fluorescence, presenting new possibilities for biofluorescence imaging. Notably, this coumarin dye also demonstrates a significant solvatochromic effect. This effect enables rapid visual discrimination among multiple sets of structurally and chemically similar organic solvents. Consequently, the dye holds promise as a versatile solvatochromic fluorescent probe for a broad spectrum of applications.

Synthesis of inverse push-pull coumarin dyes and their application as solvatochromic probes and labelling agents for bacterial cell membranes

Coumarin derivatives have been widely employed across a great number of applications, such as laser dyes, bioimaging or chemossensors, owing to their strong fluorescence and high-quantum yields. These properties are highly tunable through structural modification, namely by the inclusion of electron-withdrawing groups (EWG) and electron-donating groups (EDG) at positions 3 and 7 of the coumarin scaffold, respectively. Comparatively, there aren't many reports of EDGs being employed at position 3, prompting us to investigate this new class of potential fluorophores.We undertook the synthesis of three novel ethynyl-linked aminophenyl-coumarin dyes, employing a key Sonogashira-coupling reaction to introduce the electron-rich tertiary aniline moiety. Two different amine donors were used, diphenylamino (coumarins A and B) and dimethylamino (coumarin C), as well as two alkoxy substituents, dibenzyloxy (A and C) and methylenedioxy (B), in the phenyl ring of the coumarin. All dyes show strong emission and significant positive solvatochromism. Additionally, they were employed as staining agents for the cellular membrane of S. pneumoniae, a Gram-positive bacterial pathogen. None of the three derivatives presented cytotoxic effects. Only A and B successfully labelled the bacterial membrane. In the case of B a very similar staining pattern and performance to that of standard dye Nile Red was observed, while A seemed to possess a more selective distribution.

Fluorescent Solvatochromic Probes for Long-Term Imaging of Lipid Order in Living Cells.

High-resolution spatio-temporal monitoring of the cell membrane lipid order provides visual insights into the complex and sophisticated systems that control cellular physiological functions. Solvatochromic fluorescent probes are highly promising noninvasive visualization tools for identifying the ordering of the microenvironment of plasma membrane microdomains. However, conventional probes, although capable of structural analysis, lack the necessary long-term photostability required for live imaging at the cellular level. Here, an ultra-high-light-resistant solvatochromic fluorescence probe, 2-N,N-diethylamino-7-(4-methoxycarbonylphenyl)-9,9-dimethylfluorene (FπCM) is reported, which enables live lipid order imaging of cell division. This probe and its derivatives exhibit sufficient fluorescence wavelengths, brightness, polarity responsiveness, low phototoxicity, and remarkable photostability under physiological conditions compared to conventional solvatochromic probes. Therefore, these probes have the potential to overcome the limitations of fluorescence microscopy, particularly those associated with photobleaching. FπCM probes can serve as valuable tools for elucidating mechanisms of cellular processes at the bio-membrane level.

Solvatochromic Buffering Fluorescent Probe Resolves the Lipid Transport and Morphological Changes during Lipid Droplet Fusion by Super-Resolution Imaging.

The varied functions of lipid droplets, which encompass the regulation of lipid and energy homeostasis, as well as their association with the occurrence of various metabolic diseases, are intricately linked to their dynamic properties. Super-resolution imaging techniques have emerged to decipher physiological processes and molecular mechanisms on the nanoscale. However, achieving long-term dynamic super-resolution imaging faces challenges due to the need for fluorescent probes with high photostability. This paper introduces LD-CF, a "buffering probe" for imaging lipid droplet dynamics using structured illumination microscopy (SIM). The polarity-sensitive LD-CF eliminates background fluorescence with a "cyan filter" strategy, enabling wash-free imaging of lipid droplets. In the fluorescent "off" state outside droplets, the probes act as a "buffering pool", replacing photobleached probes inside droplets and enabling photostable long-term SIM imaging. With this probe, three modes of lipid droplet fusion were observed, including the discovery of fusion from large to small lipid droplets. Fluorescence intensity tracking also revealed the direction of lipid transport during the lipid droplet fusion.

Polarity of Organic Solvent/Water Mixtures Measured with Reichardt’s B30 and Related Solvatochromic Probes—A Critical Review

The UV/Vis absorption energies (νmax) of different solvatochromic probes measured in co-solvent/water mixtures are re-analyzed as a function of the average molar concentration (Nav) of the solvent composition compared to the use of the mole fraction. The empirical ET(30) parameter of Reichardt’s dye B30 is the focus of the analysis. The Marcus classification of aqueous solvent mixtures is a useful guide for co-solvent selection. Methanol, ethanol, 1,2-ethanediol, 2-propanol, 2-methyl-2-propanol, 2-butoxyethanol, formamide, N-methylformamide (NMF), N,N-dimethylformamide (DMF), N-formylmorpholine (NFM), 1,4-dioxane and DMSO were considered as co-solvents. The ET(30) values of the binary solvent mixtures are discussed in relation to the physical properties of the co-solvent/water mixtures in terms of quantitative composition, refractive index, thermodynamics of the mixture and the non-uniformity of the mixture. Significant linear dependencies of ET(30) as a function of Nav can be demonstrated for formamide/water, 1,2-ethanediol/water, NMF/water and DMSO/water mixtures over the entire compositional range. These mixtures belong to the group of solvents that do not enhance the water structure according to the Marcus classification. The influence of the solvent microstructure on the non-linearity ET(30) as a function of Nav is particularly clear for alcohol/water mixtures with an enhanced water structure.

Use of DFT Calculations as a Tool for Designing New Solvatochromic Probes for Biological Applications

The intramolecular charge transfer behavior of push–pull dyes is the origin of their sensitivity to environment. Such compounds are of interest as probes for bioimaging and as biosensors to monitor cellular dynamics and molecular interactions. Those that are solvatochromic are of particular interest in studies of lipid dynamics and heterogeneity. The development of new solvatochromic probes has been driven largely by the need to tune desirable properties such as solubility, emission wavelength, or the targeting of a particular cellular structure. DFT calculations are often used to characterize these dyes. However, if a correlation between computed (dipole moment) and experimentally measured solvatochromic behavior can be established, they can also be used as a design tool that is accessible to students. Here, we examine this correlation and include case studies of the effects of probe modifications and conformation on dipole moments within families of solvatochromic probes. Indeed, the ground state dipole moment, an easily computed parameter, is correlated with experimental solvatochromic behavior and can be used in the design of new environment-sensitive probes before committing resources to synthesis.

Solvatochromism in Solvent Mixtures: A Practical Solution for a Complex Problem

Many reactions are carried out in solvent mixtures, mainly because of practical reasons. For example, E2 eliminations are favored over SN2 substitutions in aqueous organic solvents because the bases are desolvated. This example raises the question: how do we chose binary solvents to favor reaction outcomes? This important question is deceptively simple because it requires that we understand the details of all interactions within the system. Solvatochromism (solvent-dependent color change of a substance) has contributed a great deal to answer this difficult question, because it gives information on the interactions between solvents, solute-solvent, and presumably transition state-solvent. This wealth of information is achieved by simple spectroscopic measurements of selected (solvatochromic) substances, or probes. An important outcome of solvatochromism is that the probe solvation layer composition is almost always different from that of bulk mixed solvent. In principle, this difference can be exploited to “tune” the composition of solvent mixture to favor the reaction outcome. This minireview addresses the use of solvatochromic probes to quantify solute-solvent interactions, leading to a better understanding of the complex effects of solvent mixtures on chemical phenomena. Because of their extensive use in chemistry, we focus on binary mixtures containing protic-, and protic-dipolar aprotic solvents.

Tetrazine-derived chromones as conditionally activated solvatochromic fluorescent probes for dual imaging of lipid droplets and mitochondria

Tetrazine-derived chromones were developed as conditionally activated solvatochromic fluorescent probes and applied for LD and mitochondrial imaging.