Mitochondria-Directing Fluorogenic Probe: An Efficient Amyloid Marker for Imaging Lipid Metabolite-Induced Protein Aggregation in Live Cells and Caenorhabditis elegans.

The design and development of optical probes for sensing neurotoxic amyloid fibrils are active and important areas of research and are undergoing continuous advancements. In this paper, we have synthesized a red emissive styryl chromone-based fluorophore (SC1) for fluorescence-based detection of amyloid fibrils. SC1 records exceptional modulation in its photophysical properties in the presence of amyloid fibrils, which has been attributed to the extreme sensitivity of its photophysical properties toward the immediate microenvironment of the probe in the fibrillar matrix. SC1 also shows very high selectivity toward the amyloid-aggregated form of the protein as compared to its native form. The probe is also able to monitor the kinetic progression of the fibrillation process, with comparable efficiency as that of the most popular amyloid probe, Thioflavin-T. Moreover, the performance of SC1 is least sensitive to the ionic strength of the medium, which is an advantage over Thioflavin-T. In addition, the molecular level interaction forces between the probe and the fibrillar matrix have been interrogated by molecular docking calculations which suggest the binding of the probe to the exterior channel of the fibrils. The probe has also been demonstrated to sense protein aggregates from the Aß-40 protein, which is known to be responsible for Alzheimer's disease. Moreover, SC1 exhibited excellent biocompatibility and exclusive accumulation at mitochondria which allowed us to successfully demonstrate the applicability of this probe to detect mitochondrial-aggregated protein induced by an oxidative stress indicator molecule 4-hydroxy-2-nonenal (4-HNE) in A549 cell lines as well as in a simple animal model like Caenorhabditis elegans. Overall, the styryl chromone-based probe presents a potentially exciting alternative for the sensing of neurotoxic protein aggregation species both in vitro as well as in vivo.

Perylene-Based Smart Fluoroprobe with Dual Function: Ratiometric Response toward Hazardous Organic Peroxides and Pure White Light Generation via Self-Assembly.

In this report, we have designed and synthesized a perylene-based smart fluoroprobe (PBE) in which the perylene core has been functionalized with the boronate group at the peri-position. PBE shows a very fast and ratiometric response toward harmful organic peroxides (OPs) generated in old ethereal solvents via auto-oxidation. The response toward OPs takes place with a visible color change from green to yellow, which could be easily observed with the naked eye. The reaction between PBE and OPs involves the cleavage of the boronate group and its consequent conversion into the -OH group. The response of PBE toward OPs was monitored using UV-vis absorption, fluorescence emission, IR spectroscopy, and mass spectrometry. Additionally, we have also explored the self-assembly of PBE in an organic-aqueous solvent mixture, which shows pure white light emission (WLE) with the CIE coordinates (0.33, 0.33) in a 50% dimethyl sulfoxide-water mixture. This work clearly reveals that PBE fluoroprobe can be employed for sensitive detection of hazardous OPs present in old ethereal solvents. Moreover, the ability of PBE to generate the perfect pure WLE makes it a potential candidate for application in organic light-emitting devices.

Trivalent metal ion sensor enabled bioimaging and quantification of vaccine-deposited Al3+ in lysosomes.

Extracellular metallic debris is deposited into the well-known 'recycle bins' of the cells named lysosomes. The accumulation of unwanted metal ions can cause dysfunction of hydrolyzing enzymes and membrane rupturing. Thus, herein, we synthesized rhodamine-acetophenone/benzaldehyde derivatives for the detection of trivalent metal ions in aqueous media. In solution, the synthesized probes exhibited a 'turn-on' colorimetric and fluorometric response upon complexation with trivalent metal ions (M3+). Mechanistically, M3+ chelation enables the appearance of a new emission band at approximately 550 nm, which verifies the disruption of the closed ring and the restoration of conjugation on the xanthene core in rhodamine 6G derivatives. Exclusive localization of the biocompatible probes at the lysosomal compartment favored the quantification of deposited Al3+. Moreover, the novelty of the work lies in the detection of Al3+ deposited in the lysosome that originated from hepatitis B vaccines, which shows their efficiency for near future in vivo applications.

A perylenemonoimide-based fluorescent probe: ultrasensitive and selective tracing of endogenous peroxynitrite in living cells.

Peroxynitrite (ONOO-), a highly reactive species, plays a key role in various physiological and pathological processes. Herein, a red-emitting fluorescent reporter perylenemonoimide-boronate ester (PMI-BE) was synthesized and utilized for ultrasensitive detection of ONOO-. The unique feature of PMI-BE is its nanomolar sensitivity with high selectivity towards ONOO-. Moreover, PMI-BE also detects endogenously generated ONOO- in live cells.

Intracellular Physical Properties with Small Organic Fluorescent Probes: Recent Advances and Future Perspectives.

The intracellular physical parameters i. e., polarity, viscosity, fluidity, tension, potential, and temperature of a live cell are the hallmark of cellular health and have garnered immense interest over the past decade. In this context, small molecule organic fluorophores exhibit prominent useful properties including easy functionalizability, environmental sensitivity, biocompatibility, and fast yet efficient cellular uptakability which has made them a popular tool to understand intra-cellular micro-environmental properties. Throughout this discussion, we have outlined the basic design strategies of small molecules for specific organelle targeting and quantification of physical properties. The values of these parameters are indicative of cellular homeostasis and subtle alteration may be considered as the onset of disease. We believe this comprehensive review will facilitate the development of potential future probes for superior insight into the physical parameters that are yet to be quantified.

Biocompatible Fluorescent Probe for Selective Detection of Amyloid Fibrils.

The misfolding and aggregation of proteins leading to amyloid formation has been linked to numerous diseases, necessitating the development of tools to monitor the fibrillation process. Here, we report an intramolecular charge transfer (ICT) dye, DMNDC, as an alternative to thioflavin-T (ThT), most commonly used for monitoring amyloid fibrils. Using insulin as a model protein, we show that DMNDC efficiently reports on the kinetics of fibril formation. An approximately 70 nm hypsochromic shift along with a large enhancement in emission intensity was observed upon binding of DMNDC to protein fibrils. The aggregation kinetics of insulin were not significantly affected in the presence of DMNDC, suggesting that DMNDC does not inhibit insulin aggregation. Additionally, the efficient cellular internalization and low toxicity of DMNDC make it highly suited for sensing and imaging of amyloid fibrils in the complex biological milieu.

Rapid on-site and naked-eye detection of common nitro pesticides with ionic liquids.

Rapid 'in-field' detection of environmentally hazardous organophosphorus and nitro-containing pesticides is highly essential due to the lethal effects caused by the inhibition of the activity of acetylcholinesterase (AChE). In our present study, we demonstrate a novel sensing approach for the simultaneous analysis of five widely used pesticides (methyl parathion, pendimethalin, dicloran, trifluralin, and PCNB) based on the Meisenheimer complex formation between polynitro aromatic compounds (pesticides) and a nucleophile. This colorimetric determination of pesticides involves the use of an ionic liquid, tetrabutylammonium hydroxide (TBAOH), as the nucleophile, which is titrated against different concentrations of pesticides. The addition of TBAOH to the solutions of pesticides results in the formation of intensely colored complexes, which are visualized using UV-vis and NMR spectroscopies allowing the identification of new bands and peaks corresponding to the formation of Meisenheimer complexes. The limit of detection (LOD) for targeted pesticides was found to be in the range of 0.67-10 µM. Furthermore, the practical application of this method is demonstrated by developing different paper-based sensors. Therefore, the strategy proposed here not only serves as a valuable tool that allows unskilled people to detect hazardous pesticides in agricultural products 'on-site' but also offers a fast and convenient protocol for the identification of dangerous nitro-containing polyaromatic groups like nitro explosives.

Visible light-controlled NO generation for photoreceptor-mediated plant root growth regulation.

Nitric oxide (NO) is an essential redox-signaling molecule free radical, contributes a significant role in a diverse range of physiological processes. Photo-triggered NO donors have significant potential compared to other NO donors because it releases NO in the presence of light. Hence, an efficient visible light-triggered NO donor is designed and synthesized by coupling 2,6-dimethyl nitrobenzene moiety at the peri-position of 1, 8-naphthalimide. The NO-releasing ability is validated using various spectroscopic techniques, the photoproduct is characterized, and finally, the NO generation quantum yield is also determined. Furthermore, the photo-generated NO has been employed to Arabidopsis thaliana as a model plant to examine the effect of photoreceptor-mediated NO uptake on plant root growth regulation molecule.

Visible-Light-Triggered Generation of Ultrastable Radical Anion from Nitro-substituted Perylenediimides.

An efficient method for visible-light-triggered generation of radicals from mono- and dinitro-substituted perylenediimide derivatives is developed. UV-vis-NIR and electron paramagnetic resonance measurements were carried out to confirm the formation of radicals. Most importantly, these radical anions were remarkably stable for several months. Subsequently, the reversible nature of anions was validated by both chemical and spectroelectrochemical methods for applications in electrochromic materials.

Rationally Designed Solvatochromic Fluorescent Indoline Derivatives for Probing Mitochondrial Environment.

A new class of solvatochromic, robust, and multifunctional fluorescent probes derived from indoline is presented. Specificity of mitochondria targeting was achieved and utilized for probing polarity under normal and apoptotic conditions. A large Stokes shift, high quantum yield, thermal, photochemical, and pH stability, tolerance to buffer compositions, and a bioconjugation tool-kit make it a promising candidate for live-cell fluorescence imaging.

Helicity-Dependent Regiodifferentiation in the Excited-State Quenching and Chiroptical Properties of Inward/Outward Helical Coumarins.

Influence of helicity on the excited-state as well as chiroptical properties of two sets of regiohelical coumarins that are differentiated by "inward" and "outward" disposition of the pyran-2-one ring has been investigated. A subtle difference in the helicities manifests in divergent excited-state properties and significant differences in the dipole moments. The latter permit heretofore unprecedented regiodifferentiation in the O-H⋅⋅⋅O hydrogen-bond assisted electron-transfer quenching by phenols. Furthermore, the enantiopure hexahelical coumarins exhibit strong Cotton effects and lend themselves to a very high differentiation in the specific rotations and anisotropic dissymmetry factors. The specific rotation observed for 6-in turns out being the highest of the values reported for all hexahelicenes reported so far.

Excited-state properties of fluorenones: influence of substituents, solvent and macrocyclic encapsulation.

A comprehensive investigation of the photophysics of a broad set of fluorenones substituted with methoxy groups at different positions brings out the importance of the location of substituents on the fluorenone core in modulating fluorescence and radiationless deactivation by way of modification of the singlet-excited state energy and its character. While the substituents at para positions are found to affect neither the fluorescence quantum yield nor the lifetime, those at meta positions are found to significantly modify the latter. A cumulative effect is observed for the substituents in that the nonradiative decay (knr) becomes progressively dominant with an increasing number of meta-methoxy substituents. For example, the trimethoxy substitution in 2,4,7-trimethoxyfluorenone (8) is found to increase knr by ca. 30 fold relative to that of the parent fluorenone (1) in a polar aprotic solvent such as acetonitrile. The predominance of nonradiative decay (knr) is rationalized from stabilization of the singlet-excited state via partial charge transfer from meta-methoxy substituents to the carbonyl group. Accordingly, a nice correlation is observed for the nonradiative (knr) rate constants versus singlet-excitation energies derived from fluorescence emission maxima of all fluorenones in acetonitrile. The macrocyclic host cucurbit[7]uril (CB7) is found to not only enhance the fluorescence of the parent fluorenone (1) significantly, but also increase the singlet lifetime considerably. Based on the changes observed in the absorption spectra and the lifetimes determined, a 1:1 host­guest complex has been proposed with CB7. The fluorescence lifetime observed in the presence of CB7 suggests that the hydrophobic fluorenone (1) can be employed as a probe to report on a polar microenvironment shielded from hydrogen bonding interactions in a polar protic solvent.

Dual role far red fluorescent molecular rotor for decoding the plasma membrane and mitochondrial viscosity.

The dysfunctions in the mitochondria are associated with various pathological conditions like neurodegeneration, metabolic disorder, and cancer, leading to dysregulated cell death. Here, we have designed and synthesized a julolidine-based molecular rotor (JMT) to target mitochondria with far-red emission accounting for mitochondrial dysfunction. JMT showed viscosity sensitivity with 160-fold enhancement in fluorescence intensity. The origin of the dark state in a lower viscous environment was investigated through density functional calculations. We have employed JMT to monitor mitochondrial dysfunction induced by nystatin using confocal and fluorescence lifetime imaging microscopy. Further, we investigated mitochondrial abnormalities under inflammatory conditions triggered by lipopolysaccharide in live HeLa cells. The cellular uptake mechanisms of JMT were studied using various endocytosis inhibitors. Moreover, we reported tracking small fluorescent molecule switching from mitochondria to the plasma membrane upon introducing mitochondrial depolarizer in cells. On treating the mitochondria potential uncoupler, JMT relocates to the cell membrane and can be utilized for understanding the interplay between mitochondria and cell membranes. Moreover, JMT was applied to stain the RBC plasma membrane isolated from human blood.

Review of NIR-responsive ''Smart'' carriers for photothermal chemotherapy.

This review focuses on the versatile applications of near-infrared (NIR)-responsive smart carriers in biomedical applications, particularly drug delivery and photothermal chemotherapy. These carriers demonstrate multi-responsive theranostics capabilities, including pH-dependent drug release, targeted delivery of chemotherapeutics, heat-mediated drug release, and photothermal tumor damage. Biological samples are transparent to NIR light with a suitable wavelength, and therefore, NIR light is advantageous for deep-tissue penetration. It also generates sufficient heat in tissue samples, which is beneficial for on-demand NIR-responsive drug delivery in vivo systems. The development of biocompatible materials with sufficient NIR light absorption properties and drug-carrying functionality has shown tremendous growth in the last five years. Thus, this review offers insights into the current research development of NIR-responsive materials with therapeutic potential and prospects aimed at overcoming challenges to improve the therapeutic efficacy and safety in the dynamic field of NIR-responsive drug delivery.

Review on Lysosomal Metal Ion Detection Using Fluorescent Probes.

Metal ions are indispensable and play an important role in living systems. Metal ions coordinated to metalloenzymes pocket activate the bound substrate and labile metal ions maintaining the ionic balance. The amount of metal ions present in various subcellular compartments of the cells is highly regulated for maintaining cellular homeostasis. An imbalance in the metal ion concentration is related to several diseases and results in serious pathological conditions. Mostly the internalized metal ions are processed in the lysosomal compartment of the cell. A delicate regulation of metal ions in the lysosomal compartment can modulate the lysosomal pH and inhibit hydrolytic enzymes, which ultimately causes lysosomal storage disorders. In the past decade, the understanding and regulation of lysosomal metal ions based on fluorometric methods have gained significant attention. In this review, we have comprehensively summarized the development of various fluorescent reporters over the past five years for a selective and sensitive estimation of lysosomal metal ion concentration. We believe this consolidated and timely review will help researchers working in the areas associated with lysosomal metal ions.

A highly efficient, selective, reversible and ultra-sensitive fluorescence "Turn-ON" chemosensor for aluminium ions by a novel Schiff base.

The synthesis of a Schiff base-based chemosensor, denoted as H6L, was accomplished through the condensation reaction of Isophthalohydrazide and 2,6-dihydroxybenzaldehyde in an ethanol solvent. The resulting compound was further characterized using 1H and 13C nuclear magnetic resonance (NMR) spectroscopy, as well as high-resolution mass spectrometry (HRMS). Extensive research has been conducted on several facets of metal sensing phenomena, revealing that the Schiff base H6L demonstrates discerning and expeditious fluorescence sensing characteristics specifically towards Al (III) in acetonitrile. The purported method detects Al (III) can be ascribed to the suppression of photo-induced electron transfer (PET) and the enhanced chelation-induced fluorescence (CHEF). The stoichiometry of metal-ligand complexes (2:1) was determined using Job's plots titrations, HRMS and subsequently confirmed using NMR titration studies. The H6L sensors demonstrated remarkable fluorescence sensing capabilities in acetonitrile, with a low detection limit (LOD) of 0.44 µM. This LOD is suitably low for the detection of Al3+, which is commonly found in many environmental and biological systems. Fluorescence lifetime measurement provides additional evidence of complexation of H6L with Al (III). The reversibility of the sensor was demonstrated through the introduction of pyrophosphate (PPi), which forms a complex with aluminium ions, thereby releasing the chemo sensor for subsequent utilization. The findings suggest that H6L has the potential to serve as a viable probe for the detection and identification of Al3+ ions.

A small-molecule probe to decipher stress-induced ER microenvironments and ER-Golgi communication.

Cellular stress is a crucial factor in regulating and maintaining both organismal and microenvironmental homeostasis. It induces a response that also affects the micropolarity of specific cellular compartments, which is essential for early disease diagnosis. In this contribution, we present a quantitative study of micropolarity changes inside the endoplasmic reticulum (ER) during the G1/S and G2/M phases, using a biocompatible small-molecule fluorophore called ER-Oct. This probe is selectively driven to the ER by its hydrophobicity, and it has the fastest diffusion properties among a series of analogous probes. We found that induced ER stress caused cell cycle arrests leading to an increase in ER micropolarity which is well supported by lambda scanning experiments and fluorescence lifetime imaging microscopy (FLIM) as well. ER-Oct is a versatile staining agent that could effectively stain the ER in various living/fixed mammalian cells, isolated ER, Caenorhabditis elegans, and mice tissues. Furthermore, we used this probe to visualize a well-known biological event, ER to Golgi transport, by live-cell fluorescence microscopy. Our exhaustive investigation of micropolarity using ER-staining dye provides a new way to study ER stress, which could provide a deeper understanding of proteostasis in model systems and even in fixed patient samples.

Ultrafast symmetry-breaking charge separation in Perylenemonoimide-embedded multichromophores: impact of regioisomerism.

Symmetry-breaking charge separation (SB-CS) has recently evolved as an emerging concept offering its potential to the latest generation of organic photovoltaics. However there are several concerns that need to be addressed to reach the state-of-the-art in SB-CS chemistry, for instance, the desirable molecular geometry, interchromophoric distance and extent of electronic coupling. To shed light on those features, it is reported herein, that ortho-functionalized perylene monoimide (PMI) constituted regioisomeric dimer and trimer derivatives with varied molecular twisting and electronic conjugation have been synthesized. In steady-state photophysical studies, all the dimers and trimer derivatives exhibit a larger bathochromic shift in the emission spectra and a significant reduction of fluorescence quantum yield in polar DMF. Among the series of multichromophores, ortho- and self-coupled dimers display the strikingly different optical feature of SB-CS with a very fast charge separation rate (τCS = 80.2 ps) upon photoexcitation in DMF, which is unveiled by femtosecond transient absorption (fs-TA) studies. The SB-CS for two dimers is well-supported by the formation of PMI˙+ and PMI˙- bands in the fs-TA spectra. Further analysis of fs-TA data revealed that, among the other multichromophores the trimer also exhibits a clear charge separation, whereas SB-CS signatures are less prominent, but can not be completely disregarded, for the meta- and para-dimers. Additionally, the charge separation dynamics of those above-mentioned PMI derivatives are devoid of a kinetically favorable excimer or triplet formation. The evidence of a profound charge transfer phenomenon in the ortho-dimer is characterized by density functional theory (DFT) calculations on excited state electronic structures. The excitonic communications in the excited state electronic arrangements unravel the key role of dihedral twisting in SB-CS. The thermodynamic feasibility of CS (ΔGCS) and activation barrier (ΔG≠) of the derivatives in DMF are established from the Rehm-Weller equation and Marcus's theory, respectively. This work is an in-depth study of the effect of mutual orientation of PMIs and regioisomerism in determining sustainable guidelines for using SB-CS.

Tracking endoplasmic reticulum viscosity during ferroptosis and autophagy using a molecular rotor probe.

Ferroptosis is a unique non-apoptotic cell death process associated with endoplasmic reticulum (ER) stress-related diseases. We have designed and synthesized a far-red emitting and ER targetable viscosity-sensitive fluorophore to track ER-phagy. Furthermore, the ER viscosity alteration during the ferroptosis process was investigated via intensity and lifetime-based spectroscopy and microscopy.

Bioimaging of labile lysosomal iron through naphthalimide-based fluorescent probe.

Lysosomal labile iron detection is immensely important as it is related to various diseases like Alzheimer's disease, Huntington's disease, Parkinson's disease, and cell apoptosis like ferroptosis. The fluorescent-based detection methods are preferred due to their sensitive, non-invasive, and spatial-temporal detection in biological samples. However, this remains a great challenge due to the lysosomal compartment being acidic alters the photophysical properties of the probe. Herein, we have rationally designed and synthesized multi-component naphthalimide-based fluorescent marker with preferred optical properties and bio-compatibility for selective detection of labile iron present in the lysosomal compartment. The synthesized probe was characterized structurally and optically by NMR, mass spectrometry, UV-visible, and fluorescence spectroscopy. The developed probe with an appropriate linking strategy turns out to be tolerant to fluorescence alternation in lysosomal pH. The probe exhibits great selectivity and high sensitivity for Fe(III) with limit of detection of 0.44 µM and is also able to detect Fenton-type reactions. Further, the probe has been successfully applied for lysosomal imaging and detecting labile Fe(III) present in the lysosomal lumen of the live cells.