Dual Colors Research Articles

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.

Exquisite visualization of mitophagy and monitoring the increase of lysosomal micro-viscosity in mitophagy with an unusual pH-independent lysosomal rotor

BackgroundMitophagy plays indispensable roles in maintaining intracellular homeostasis in most eukaryotic cells by selectively eliminating superfluous components or damaged organelles. Thus, the co-operation of mitochondrial probes and lysosomal probes was presented to directly monitor mitophagy in dual colors. Nowadays, most of the lysosomal probes are composed of groups sensitive to pH, such as morpholine, amine and other weak bases. However, the pH in lysosomes would fluctuate in the process of mitophagy, leading to the optical interference. Thus, it is crucial to develop a pH-insensitive probe to overcome this tough problem to achieve exquisite visualization of mitophagy. ResultsIn this study, we rationally prepared a pH-independent lysosome probe to reduce the optical interference in mitophagy, and thus the process of mitophagy could be directly monitored in dual color through cooperation between IVDI and MTR, depending on Förster resonance energy transfer mechanism. IVDI shows remarkable fluorescence enhancement toward the increase of viscosity, and the fluorescence barely changes when pH varies. Due to the sensitivity to viscosity, the probe can visualize micro-viscosity alterations in lysosomes without washing procedures, and it showed better imaging properties than LTR. Thanks to the inertia of IVDI to pH, IVDI can exquisitely monitor mitophagy with MTR by FRET mechanism despite the changes of lysosomal pH in mitophagy, and the reduced fluorescence intensity ratio of green and red channels can indicate the occurrence of mitophagy. Based on the properties mentioned above, the real-time increase of micro-viscosity in lysosomes during mitophagy was exquisitely monitored through employing IVDI. Significance and noveltyCompared with the lysosomal fluorescent probes sensitive to pH, the pH-inert probe could reduce the influence of pH variation during mitophagy to achieve exquisite visualization of mitophagy in real-time. Besides, the probe could monitor the increase of lysosomal micro-viscosity in mitophagy. So, the probe possesses tremendous potential in the visualization of dynamic changes related to lysosomes in various physiological processes.

Salvia guidongensis sp. nov.: unraveling a critical evolutionary link in East Asian Salvia from Central China integrating morphology, phylogeny, and plastid genomics.

Salvia L., representing the largest genus within the mint family, is noted for its global distribution of approximately 1000 species, with East Asia, and particularly China, recognized as a critical center of diversity for the genus. Our research was conducted through extensive fieldwork in Guidong County, Hunan Province, China, where we identified a previously undescribed species of Salvia. The identification process involved detailed morphological observations, phylogenetic analyses, and plastid genomics. The newly discovered species, Salvia guidongensis, exhibits unique characteristics not commonly observed in the East Asian lineage of Salvia, including dual floral colors within natural populations-either pale purple or pale yellow. Morphologically, while it shares similarities with members of sect. Glutinaria, S. guidongensis is distinct in its floral morphology, stature, and specific foliar traits. Phylogenetic analysis places S. guidongensis in a unique clade within the East Asian lineage of Salvia, suggesting it may serve as an important evolutionary link. Additionally, we explored the plastome features of S. guidongensis, comparing them with those of closely related species. The discovery of S. guidongensis not only entriches the taxonomic tapestry of Salvia but also provides critical insights into the biogeography and evolutionary pathways of the genus in East Asia. By integrating morphological and molecular data, we validate the novel status of S. guidongensis and highlight its significance in bridging taxonomic and evolutionary gaps within Sect. Glutinaria of Salvia.

Single Fluorescent Probe for Multiple Tasks: Illuminating Lipid Droplets and Lysosomes in Dual Channels and Distinguishing Autophagy and Apoptosis.

Lipid droplets (LDs) and lysosomes play key roles in autophagy and cell apoptosis, and the discriminative visualization of the two organelles and simultaneously of autophagy and apoptosis is very helpful to understand their internal relationships. However, fluorescent probes that can concurrently achieve these tasks are not available currently. Herein, we delicately fabricate a robust probe CAQ2 for multiple tasks: illumination of LDs and lysosomes in dual emission colors as well as discriminative visualization of cell apoptosis and autophagy. The probe exhibited both lipophilic and basic properties and displayed different emission colors in neutral and protonated forms; thus, LDs and lysosomes emitted blue and red fluorescence colors, respectively. Because of the lysosomal acidification during autophagy, CAQ2 detected autophagy with evidently enhanced red emission. Because of the lysosomal alkalization during apoptosis, CAQ2 imaged apoptosis with a drastically decreased red fluorescence intensity. With the robust probe, the autophagy under starvation and lipidless conditions was visualized, and the apoptosis induced by H2O2, ultraviolet (UV) irradiation, and rotenone treatment was successfully observed. The efficient detoxification of Na2S against rotenone treatment was successfully revealed.

Construction of a Dual-Emissive Probe for Discriminative Visualization of Lysosomal and Mitochondrial Dysfunction.

Discriminatively visualizing mitochondrial and lysosomal dysfunction is crucial for an in-depth understanding of cell apoptosis regulation and relative biology. However, fluorescent probes for the separate visualization of lysosomal and mitochondria damages have not been reported yet. Herein, we have constructed a fluorescent probe [2-(4-hydroxystyryl)-1,3,3-trimethyl-3H-indol-1-ium iodide (HBSI)] for labeling mitochondria and lysosomes in dual emission colors and discriminatively imaging mitochondrial and lysosomal damage in two different sets of fluorescent signals. In living cells, HBSI targeted both lysosomes and mitochondria to give green and red emission, respectively. During mitochondrial damages, HBSI immigrated into lysosomes, and the red emission decreased. During lysosomal damage, HBSI immigrated into mitochondria, and the green emission decreased. With the robust probe, the different damaging sequences of mitochondria and lysosomes under different amounts of H2O2 and chloral hydrate have been revealed. The sequential damage of lysosomes and mitochondria during cell apoptosis induced by rotenone, paclitaxel, and colchicine has been discovered. Furthermore, the regulation of mitochondria, lysosome, and their interplay during autophagy was also observed with the probe.

Development of a high polarity-sensitive fluorescent probe for visualizing the lipid droplets and endoplasmic reticulum with dual colors in living cells

Lipid droplets (LDs) are unique organelles that control the lipid metabolism in cells. It has been identified that the generations of LDs derive from endoplasmic reticulum (ER) and they have closely related with amount of cellular activities for maintaining homeostasis. To further explore the detail interactions between LDs and ER, we have developed a novel polarity-sensitive fluorescent probe LP with distinct D-π-A-π-D framework and applied it to imaging LDs and ER with dual colors at the same time. Probe LP showed well red-shifted emissions with the increase fraction of water in the 1,4- dioxane due to ICT process. In biological imaging, probe LP could visualize LDs and ER with green and red fluorescence separately. Besides, the dynamic behaviors of LDs and ER were achieved using LP during the oleic acids and starvation stimulations. Therefore, probe LP is a valuable molecular tool for investigating the relationships of LDs and ER in various cellular activities.

Polarization-Sensitive Structural Colors Based on Anisotropic Silicon Metasurfaces

Structural colors based on all-dielectric metasurfaces hold great promise for a wide range of applications, including high-density optical storage, ultra-high-resolution 3D displays, imaging security certification, and so on. However, achieving dynamic tunable structural color with a compact and simple Si platform remains a great challenge. Here, we propose a dynamic tunable structural coloration with polarization-sensitive metasurfaces consisting of arrays of Si elliptical nanopillars, enabling full-colored images to be displayed and switched through the control of the polarization of incident light. A distinct feature of our design is that the color phase is independent of the viewing angle, which is fundamental for real applications. Moreover, we demonstrated that dual and multiple colors can be obtained by varying the angle of either the polarizer or the analyzer. Our scheme provides a simple yet general approach for potential applications in the fields of virtual reality, ultra-high-resolution 3D displays, and high-density information storage.

Microfluidics-Assisted Fabrication of Dual Stopband Photonic Microcapsules and Their Applications for Anticounterfeiting.

The assembly of two different kinds of colloidal particle-based photonic structures into an individual micro-object can achieve multifunctionality. In this study, core–shell photonic microcapsules with dual structural colors and photonic stop bands were prepared through a standard microfluidic technique. Photocurable resin suspension of silica nanoparticles and an aqueous suspension of nanogels were used as shell and core parts of microcapsules, respectively. The structural colors of shells and cores can be tuned by adjusting the concentrations of silica nanoparticles and soft nanogels in their corresponding suspensions. The individual microcapsules possess two distinct stop bands when the two suspensions are combined appropriately. Remarkably, the color information of the core part cannot be directly viewed at a macroscopic level (such as visual inspection) but can be detected at a microscopic scale (such as optical microscopy observation). The color information hidden enables the capability for information encryption and has potentially critical applications in anti-counterfeiting, display, and other fields.

One-Pot Self-Assembly of Dual-Color Domes Using Mono-Sized Silica Nanoparticles.

Spots with dual structural colors on the skin of some organisms in nature are of tremendous interest due to the unique function of their dye-free colors. However, imitation of them requires complicated manufacturing processes, expensive equipment, and multiple predesigned building blocks. In this work, a one-pot strategy based on the phase-separation-assisted nonuniform self-assembly of monosized silica nanoparticles is developed to construct domes with dual structural colors. In drying poly(ethylene glycol)-dextran-based (PEG-DEX) droplets, monosized nanoparticles distribute nonuniformly in two compartments due to the droplet inner flow and different nanoparticle compatibility with the two phases. The dome colors are derived from the self-assembled nanoparticles and are programmable by regulating the assembly conditions. The one-pot strategy enables the preparation of multicolor using only one type of building block. With the dual-color domes, encrypted patterns with a high volume of contents are designed, showing promising applications in information delivery.

Revealing the Phase Separation in ER Membranes of Living Cells and Tissues by In Situ NIR Ratiometric Imaging

Biomembranes in the endoplasmic reticulum (ER) play indispensable roles in various bioactivities, and therefore, visualizing the phase separation in ER membranes is crucial for the studies on the fundamental biology of the ER. However, near-infrared (NIR) ratiometric imaging of the phase behaviors of the ER in living cells with different statuses and in diverse tissues has not been investigated. Herein, we developed a polarity-responsive NIR fluorescent probe (DCA) for the visualization of the phase behavior in ER membranes. The probe displayed a large Stokes shift and was highly sensitive to polarity. By direct and native fluorescence imaging at room temperature, the ERo and ERd biomembranes in the ER could be clearly distinguished by dual NIR emission colors. Oxidative damage by H2O2 and homocystein (Hcy)-induced ER stress can efficiently induce the formation of large-scale ERo domains in ER membranes. Moreover, we have also revealed that different tissues exhibited diverse phase behaviors in the ER membranes. The ER membranes in cardiac and skeletal muscle tissues showed no evident phase separation, while large-scale ERo domains existed in the ER of liver tissues and formed at the ER membranes adjacent to lipid droplets (LDs) in white adipose tissues. We expect that the probe could serve as a powerful molecular tool to promote fundamental research studies on ER membranes and relative biomedical areas.

Tuning the “critical polarity” of TICT dyes: Construction of polarity-sensitive platform to distinguish duple organelles

Polarity-sensitive dyes have demonstrated potential applications in bioimaging experiments. However, the twisted intramolecular charge transfer (TICT) mechanism to explain the polarity-responsive behavior still requires in-depth exploration. Herein, we proposed a new concept, “critical polarity”, for the in-depth explanation of TICT mechanism. The polarity below “critical polarity” was defined as “low polarity”, in which the fluorophore would present in local excited (LE) states. The polarity above “critical polarity” was defined as “high polarity”, in which the fluorophore would present in TICT states. Both proper “critical polarity” and large emission difference between LE and TICT states are required to construct desirable dual-emissive fluorophores highly sensitive to polarity. Three fluorescent dyes have been fabricated by modifying different electron donors on naphthalene imide, and S-NAT with modest electron donor demonstrated the highest sensitivity to polarity. Theoretical analysis together with the experimental results proved the validation of “critical polarity” mechanism. Finally, two fluorescent probes have been constructed based on the polarity-sensitive fluorophore. Amongst, the S-NAT could distinguish endoplasmic reticulum (ER) and lipid droplets (LDs) in dual emission colors, while S-NAT-M enabled discrimination of lysosomes and LDs in different emission colors. The two fluorescent probes with high quantum yield and low cytotoxicity could be employed to study the interaction between duple organelles, and the “critical polarity” mechanism can serve as a validate strategy to direct the construction of ratiometric dyes for polarity.

Bonding Strength Analysis of Multi-material and Multi-color Specimens Printed with Multi-extrusion Printer

This paper highlights the findings of multi-material application in 3D printed specimens. The work presents experimental results of multi-material, and multi-color-based 3D printed specimens by a multi-extrusion printer developed at the laboratory. It aimed to determine the bonding properties between the layers of various materials indicated with different colors. The samples were printed across multiple infill models and tested in the tensile and compression machine. The specimens were created with 10%, 25%, and 100% infill having single and dual colors material. Polylactic Acid (PLA), Acrylonitrile Butadiene Styrene (ABS), and Flex materials were used for printing various specimens with various colors. The layers were printed synchronously. The filaments were changed according to the predetermined algorithm. The experimental results showed that the mechanical properties of single, dual, and triple mate-rial specimens differed according to the reinforcement strategy. In addition, the mechanical property of the same material with different colors was identical.

Construction of single fluorescent probes for separately visualizing duple organelles in different emission colors

Fluorescent probes separately labelling duple organelles in different colors are powerful molecular tools to investigate the interplay between different organelles. In this work, we have presented a fluorescent dye with high sensitivity to environmental polarity, and prepared two fluorescent probes to visualize duple organelles in dual emission colors based on the platform. Probe HFM simultaneously targeted lipid droplets (LDs) and endoplasmic reticulum (ER) in living cells, and imaged the two organelles in blue and green emission, respectively. Meanwhile, HFM-M enabled visualizing LDs and lysosomes in blue and green fluorescence, respectively. Using the two newly designed fluorescent probe, the changes in LDs/ER or LDs/lysosomes during oleic acid and starvation stimuli have been visualized. We have successfully revealed that excessive alcohol can lead to the depletion of LDs and ER in living cells. We believe that the unique fluorescent dye highly sensitive to environmental polarity can serve as desirable platform for probes design. The probes HFM and HFM-M are valuable molecular tools to investigate the interplay between LDs/ER or LDs/lysosome, and facilitate the researches on relative biological fields.

Mass Spectra in $$\boldsymbol{\mathcal{N}=1SQCD}$$ with Additional Colorless Fields and Problems with the Seiberg Duality

Considered is the direct ${\cal N}=1$ SQCD-like $\Phi$-theory with $SU(N_c)$ colors and $3N_c/2< N_F<2N_c$ flavors of light quarks ${\overline Q},\,Q$. Besides, it includes $N^2_F$ additional colorless but flavored fields $\Phi_{i}^{j}$ with the large mass parameter $\mu_{\Phi}\gg\Lambda_Q$, interacting with quarks through the Yukawa coupling. In parallel, is considered its Seiberg's dual variant, i.e. the $d\Phi$-theory with $(N_F-N_c)$ dual colors, $N_F$ flavors of dual quarks ${\overline q},\,{q}$ and $N_F^2$ elementary mion fields $M^i_j\rightarrow ({\overline Q}_j Q^i)$. In considered here vacua, the quarks of both theories are in the conformal regimes at scales $\mu<\Lambda_Q$. The mass spectra are calculated in sections 4 and 5. It is shown that they are different in the direct and dual theories, in disagreement with the Seiberg hypothesis about equivalence of two such theories. Besides it is shown in the direct theory that a qualitatively new phenomenon takes place: the seemingly heavy fields $\Phi$ `return back' and there appear two additional generations of light $\Phi$-particles with small masses $\mu^{\rm pole}_{2,3}(\Phi)\ll\Lambda_Q$. In Conclusions also presented comparison of mass spectra of these two theories for such values of parameters when the direct theory is in the very strong coupling regime, while the dual one is in the weak coupling IR-free logarithmic regime. It is shown that mass spectra of these two theories are parametrically different in this case.

Gabor Filter and Moment Invariant via LDA Classifier for Skin Cancer Detection

Skin cancer may be a serious tumor. This can be clearly seen through the mature, uncommon appearance of fur pathology, which has abnormal properties in complex situations, wrinkled or uncertain perimeters, and dual colors. A small number of tulle melanomas of uncertain diameter can imitate benign moles and cannot be perceived by optical inspection. The only assumption for analyzing them is through dermoscopy as an option. Original identification and medical surgery can alternative for the patients. Within this research a detection method through image processing with various feature extraction such as Gabor filter and Hu Moment were employed and substantially improves the diagnosis performance with 97% via LDA Classifier.

SFTA and GLCM via LDA Classifier for Skin Cancer Detection

Skin cancer may be a serious tumor. This can be clearly seen through the mature, uncommon appearance of fur pathology, which has abnormal properties in complex situations, wrinkled or uncertain perimeters, and dual colors. A small number of tulle melanomas of uncertain diameter can imitate benign moles and cannot be perceived by optical inspection. The only assumption for analyzing them is through dermoscopy as an option. Original identification and medical surgery can alternative for the patients. Within this research a detection method through image processing with various feature extraction such as Gabor filter and Hu Moment were employed and substantially improves the diagnosis performance with 97% via LDA Classifier.

Design of small monomeric and highly bright near-infrared fluorescent proteins

Due to the low absorbance in the far-red (FR) and near-infrared (NIR) “optical window”, NIR fluorescent proteins (FPs) are powerful tools for deep imaging. Here, we report three new, highly bright NIR FPs termed BDFP1.8, BDFP1.8:1.8 (tandem BDFP1.8) and BDFP1.9, which evolved from a previously reported FR FP, BDFP1.6: a derivative of ApcF2 from Chroococcidiopsis thermalis sp. PCC7203. ApcF2 binds phycocyanobilin (PCB) non-covalently, while BDFPs, the derivatives of ApcF2, can bind biliverdin (BV) covalently. We identified that dimeric BDFP1.8 and monomeric BDFP1.8:1.8 have a 2.4-and 4.4-fold higher effective brightness, respectively, than iRFP720, which has the highest effective brightness among the reported NIR FPs. Monomeric DBFP1.9 (17 kDa) has one of the smallest masses among highly bright FPs in the FR and NIR regions. Enhancing the affinity between the apo-proteins and the BV chromophore is an effective method to improve the effective brightness of biliprotein FPs. Moreover, BDFP1.8 and 1.9 exhibit higher stability to temperature, pH and light than iRFP720. Finally, the highly bright NIR BDFP1.8 together with FR BDFP1.6 could effectively biolabel cells in dual colors.

Construction of the FRET Pairs for the Visualization of Mitochondria Membrane Potential in Dual Emission Colors.

Mitochondria membrane potential (MMP) play significant roles during metabolism, signaling, and other important bioevents. Visualization of MMP levels is essential for many biological researches. However, fluorescent probes for monitoring MMP levels in dual emission colors are still deficient, which greatly limited the development of relative research areas. In this work, a pair of fluorescent probes have been designed and synthesized to monitor the MMP levels in dual emission colors based on Forster resonance energy transfer (FRET) mechanism. The FRET donor (FixD) is constructed by linking a benzyl chloride group to a fluorophore with bright-green emission. The FixD could target mitochondria and be immobilized in mitochondria by linking to the thiol group of mitochondrial proteins. The FRET acceptor (LA) is designed with green absorption and deep-red emission. In live cells with high MMP levels, FixD and LA both target mitochondria, and deep-red (DR) emission could be detected with the excitation of 405 nm. Particularly, the spectral shift of fluorescence upon the decrease of MMP is up to 110 nm, which is greatly favorable for the clear observation of MMP levels. With the decrease of MMP, LA would be released from mitochondria while FixD would still be immobilized in mitochondria, and decreased DR emission and increased green fluorescence could be detected due to the absence of FRET. In this manner, the MMP levels could be monitored in dual emission colors.

Dynamically Monitoring Cell Viability in a Dual-Color Mode: Construction of an Aggregation/Monomer-Based Probe Capable of Reversible Mitochondria-Nucleus Migration.

Mitochondria and nucleus play crucial roles during cell apoptosis process. In this work, a unique fluorescent probe capable of reversible migration between mitochondria and nucleus, as well as detection of cell viability in a dual-color mode is presented. The dual-color probe targets mitochondria in healthy cells, to form aggregates with deep-red emission. It migrates into nucleus and binds to DNA to form monomers with green fluorescence during apoptosis. Interestingly, the migration is reversible dependent on cell viability, which enables the dynamic visualization of apoptosis process. With the probe, mitochondria and nucleus can be visualized in dual colors during apoptosis, and the cell viability could be monitored by the emission color and localization of the probe.

Discriminating Live and Dead Cells in Dual-Color Mode with a Two-Photon Fluorescent Probe Based on ESIPT Mechanism.

Discrimination of live and dead cells is an important task in biological, pathological, medical, and pharmaceutical studies. In this work, we have developed a novel fluorescent probe DACA that can discriminate live and dead cells in a dual-color mode for the first time. DACA can stain dead cells with blue fluorescence peaked at 440 nm, while it can also label live cells with orange emission peaked at 570 nm. Compared with one-color fluorescent probes, such a dual-color probe can efficiently avoid false positive results from cellular autofluorescence and misleading signals brought by inhomogeneous staining, and thus can supply more accurate information in biological applications. By means of DACA, the health status of tumor cells pretreated by H2O2 and ultraviolet radiation has been successfully detected and imaged. Moreover, DACA and the hydrolyzed product exhibit excellent two-photon properties. Live and dead cells, as well as the zebrafishes, have been discriminated with dual emission colors under one- and two-photon microscope. These results demonstrate that DACA is a powerful tool for dual-color distinguishing live and dead cells in vitro and in vivo.