Bioimaging Probes Research Articles

Cancer Cell Targeting Via Selective Transferrin Receptor Labeling Using Protein-Derived Carbon Dots.

Carbon dot (CD) nanoparticles offer tremendous advantages as fluorescent probes in bioimaging and biosensing; however, they lack specific affinity for biomolecules, limiting their practical applications in selective targeting. Nanoparticles with intrinsic affinity for a target have applications in imaging, cytometry, therapeutics, etc. Toward that end, we report the transferrin receptor (CD71) targeting CDs, synthesized for the first time. The formation of these particles is truly groundbreaking, as direct tuning of nanoparticle affinity was achieved by simple and careful precursor selection of a protein, which has the targeting characteristic of interest. We hypothesized that the retention of the original protein's peptides on the nanoparticle surface provides the CDs with some of the function of the precursor protein, enabling selective binding to the protein's receptor. This was confirmed with FTIR (Fourier transform infrared) data and subsequent affinity-based cell assays. These transferrin (Tf)-derived CDs have been shown to possess an affinity for CD71, a cancer biomarker that is ubiquitously expressed in nearly every cancer cell line due to its central role mediating the uptake of cellular iron. The CDs were tested using the human leukemia cell line HL60 and demonstrated the selective targeting of CD71 and specific triggering of transferrin-mediated endocytosis via clathrin-coated pits. The particle characterization results reflect a carbon-based nanoparticle with bright violet fluorescence and 7.9% quantum yield in aqueous solution. These unpresented CDs proved to retain the functional properties of the precursor protein. Indicating that this process can be repeated for other disease biomarkers for applications ranging from biosensing and diagnostic bioimaging to targeted therapeutics.

A near-infrared emitting “off-on” fluorescent probe for bioimaging of Pd(Ⅱ) ions in living cells and mice

BackgroundThe surging consumption of palladium in modern industry has given rise to its accumulation in the ecosystem, posing conspicuous toxicity to aquatic organisms and human health. The investigation of palladium in biological systems is highly demanded for the in-depth understanding of its dynamics and behaviors. Fluorescence imaging serves as a powerful approach to assess palladium species in biological systems, and currently most of the sensing probes are applicable to living cells. Effective tracking of palladium species in living organisms is challenging, which requires sufficient hydrophilicity and imaging depth of the probes. ResultsBased on an intramolecular charge transfer (ICT) mechanism, a distyryl boron dipyrromethene (BODIPY) derivative (DISBDP-Pd) has been prepared for the near-infrared (NIR) fluorescence imaging of Pd2+ ions. Two additional methoxy triethylene glycol (TEG) chains could serve as flexible and hydrophilic moieties to enhance the aqueous solubility and cell permeability of the extended conjugate. Solution studies revealed that DISBDP-Pd exhibited a NIR fluorescence enhancement signal exclusively to Pd2+ ions (detection limit as low as 0.85 ppb) with negligible interference from Pd0 species and other closely related metal ions. Computational calculations have been performed to rationalize the binding mode and the mechanism of action. Fluorescence imaging assays have been conducted on A549 human non-small cell lung carcinoma cells and mouse models. Exhibiting negligible cytotoxicity, DISBDP-Pd demonstrated concentration-related fluorescence enhancement signals in response to Pd2+ ions in living cells and mice. SignificanceDISBDP-Pd exhibits advantages over many small molecule palladium probes in terms of satisfactory aqueous solubility, high sensitivity and selectivity, and biocompatible NIR emission property, which are particularly favorable for the sensing application in biological environments. The design strategy of this probe can potentially be adopted for the functionalization of other BODIPY probes implemented for NIR fluorescence bioimaging.

Protein corona exacerbated inflammatory response in macrophages elicited by CdTe quantum dots

Nano-bio interface is significant concern in nanomedicine. When nanoparticles (NPs) come into contact with cells, they form complexes with proteins known as protein corona (PC). Cadmium telluride quantum dots (CdTe QDs) have been applied as bioimaging probes and for macrophage theragnostic. However, the impact of protein corona on the behavior of CdTe QDs is not well understood. Macrophages play a crucial role in defending against NPs. In this study, RAW264.7 cells were used to investigated the inflammatory response in macrophages when exposed to CdTe QDs before and after PC formation in fetal bovine serum. The results indicated that protein corona polarized more macrophages towards M1 phenotype. Transcriptomics analysis revealed that PC-CdTe QDs altered a greater number of differentially expressed genes (DEGs) compared to CdTe QDs (177 and 398) at 1.0 μM in macrophages. The DEGs affected by PC-CdTe QDs contained several personalized inflammatory cytokines. The enriched pathways after PC formation included Cytokine-cytokine receptor interaction, NOD-like receptor signaling pathway, and TNF signaling pathway, etc. Furthermore, PC specifically exacerbated the overexpression of CCL2 and IL-1β proteins. Importantly, PC altered the mechanism of CdTe QD-induced pyroptosis, shifting it from activating NLRC4 to both NLRP1 and NLRP3 inflammasomes, and from cleaving GSDMD and GSDMB to GSDMB alone. Overall, protein corona exacerbated the inflammatory response induced by CdTe QDs in macrophages. This study provides valuable insight into the pro-inflammatory effect of protein corona on CdTe QDs, with implications for their use in bioimaging or macrophage theragnostic by either exploiting or eliminating this biological interface effect.

Photoswitchable coordination cages.

Stimuli-responsive behaviour is key to the design of smart materials, surfaces, nano-systems and effector molecules, allowing their application as switchable catalysts, molecular transporters, bioimaging probes or caged drugs. Supramolecular chemistry has embraced the widespread integration of photoswitches because of their precise spatiotemporal addressability and waste-free nature. In the vibrant area of discrete metal-mediated self-assembly, however, photoswitches are still rarely employed. Only recently has it been shown that embedding photoswitches into the organic backbones of coordination cages enables control of their host and material properties and thus unlocks the hitherto unexploited dynamic adaptivity of such systems. Here we discuss four cases where triggering ligand-integrated photoswitches leads to (1) control over disassembly/reassembly, (2) bi-stable switching between defined states, (3) interplay with thermal processes in metastable systems and (4) light-fuelled dissipative self-assembly. We highlight first clues concerning the relationship between fundamental photophysics and dynamic assembly equilibria and propose directions for future development.

Solid-state mechanochemistry for the rapid and efficient synthesis of tris-cyclometalated iridium(iii) complexes.

Tris-cyclometalated iridium(iii) complexes have received widespread attention as attractive prospective materials for e.g., organic light-emitting diodes (OLEDs), photoredox catalysts, and bioimaging probes. However, their preparation usually requires prolonged reaction times, significant amounts of high-boiling solvents, multistep synthesis, and inert-gas-line techniques. Unfortunately, these requirements represent major drawbacks from both a production-cost and an environmental perspective. Herein, we show that a two-step mechanochemical protocol using ball milling enables the rapid and efficient synthesis of various tris-cyclometalated iridium(iii) complexes from relatively cheap iridium(iii) chloride hydrate without the use of significant amounts of organic solvent in air. Notably, a direct one-pot procedure is also demonstrated. The present solid-state approach can be expected to inspire the development of cost-effective and timely production methods for these valuable iridium-based complexes, as well as the discovery of new phosphorescent materials, sensors, and catalysts.

Covalently hybridized carbon dots@mesoporous silica nanobeads as a robust and versatile phosphorescent probe for time-resolved biosensing and bioimaging.

Phosphorescence analyses have attracted broad attention due to their remarkable merits of the elimination of auto-fluorescence and scattering light. However, it remains a great challenge to develop novel materials with uniform size and morphology, stability, long lifetime, and aqueous-phase room temperature phosphorescence (RTP) characteristics. Herein, monodisperse and uniform RTP nanobeads were fabricated by an in situ covalent hybridization of carbon dots (CDs) and dendritic mesoporous silicon nanoparticles (DMSNs) via silane hydrolysis. The formation of Si-O-C and Si-C/N covalent bonds is beneficial for the fixation of vibrations and rotations of the luminescent centers. Specially, the nanopores of DMSNs provide a confined area that can isolate the triplet state of CDs from water and oxygen and thus ensure the occurrence of aqueous-phase RTP with an ultra-long lifetime of 1.195 s (seen by the naked eye up to 9 seconds). Through surface modifying folic acid (FA), CDs@DMSNs can serve as a probe to distinguish different cell lines that feature varying FA receptor expression levels. In addition, taking MCF-7 as the model, highly sensitive and quantitative detection (linear range: 103-106 cells per mL) has been achieved via an RTP probe. Furthermore, their potential applications in cellular and in vivo time-gated phosphorescence imaging have been proposed and demonstrated, respectively. This work would provide a new route to design CD-based RTP composites and promote their further applications in the medical and biological fields.

Rigidify styryl-pyridinium dyes to benzo[h]coumarin-based bright two-photon fluorescent probes for cellular bioimaging.

Fluorescence imaging of organelles at the cellular level is important for studying biological processes. The development of a highly emissive fluorescent probe that operates under a suitable excitation light source is a key step in high-quality fluorescence imaging. For long-term, high-fidelity fluorescence imaging of mitochondria-related cellular processes using two-photon microscopy and stimulated emission depletion microscopy, we developed a new benzocoumarin-based cationic fluorescent probe (BS-CN) that is far-red emitting, water-soluble, photostable, and very bright in cells. BS-CN showed a remarkably high quantum yield of 0.35 and a large two-photon excited fluorescence action cross-section of 76 GM, enabling the long-term tracking of mitochondria in live cells. In addition, BS-CN exhibited a certain affinity for RNA and stained nucleoli in fixed cells. A comparative assessment of the photophysical properties and bioimaging performance of benzo[h]coumarin-pyridinium and the structurally similar styryl-pyridinium (BS-MN) clearly indicated the importance of structural rigidity for fluorescence efficiency.

Luminescent metal nanoclusters and their application in bioimaging.

Owing to their unique optical properties and atomically precise structures, metal nanoclusters (MNCs) constitute a new generation of optical probe materials. This mini-review provides a brief overview of luminescence mechanisms and modulation methods of luminescent metal nanoclusters in recent years. Based on these photophysical phenomena, the applications of cluster-based optical probes in optical bioimaging and related sensing, disease diagnosis, and treatment are summarized. Some challenges are also listed at the end.

A novel dual near-infrared fluorescent probe for bioimaging and visualization of viscosity in acute alcoholic liver injury.

A dual NIR fluorescent probe Cy-ND is developed for viscosity sensing with λex/em = 766/806 nm, making it apt for biological analysis, whose response is validated through DFT and TDDFT computations. Cy-ND successfully detected viscosity changes amidst acute alcohol-induced liver injury and liver ischemia-reperfusion injury.

Multimillion Atom Simulations of Di-8-ANEPPS Chromophores Embedded in a Model Plasma Membrane: Toward the Investigation of Realistic Dyed Cell Membranes.

A multistep computational approach has been employed to study a multimillion all-atom dyed plasma membrane, with no less than 42 different lipid species spanning the major head groups and a variety of fatty acids, as well as cholesterol, with the objective of investigating its structure and dynamics, as well as its impact on the embedded di-8-ANEPPS dyes. The latter are commonly used as bioimaging probes and serve as local microscopes. So, they provide information on membrane morphology via their second harmonic nonlinear optical (NLO) responses, which have the advantage of being specific to interface regions and sensitive to the chromophore environment. In previous studies, this chromophore has only been studied in simpler membrane models, far from the complexity of real lipid bilayers, while, owing to the ever-increasing computational resources, multimillion lipid bilayers have been studied, giving access to the effects of its heterogeneity. First, using molecular dynamics (MD) simulations, it is found that the combination of lipids produces a more ordered and denser membrane compared to its homogeneous model counterparts, while the local environment of the embedded dyes becomes enriched in phosphatidylcholine. Subsequently, the second harmonic first hyperpolarizability of the probes was calculated at the TDDFT level on selected frames of MD, highlighting the influence of the lipid environment. Due to the complexity of the system, machine learning (ML) tools have been employed to establish relationships between the membrane structural parameters, the orientation of the probes, and their NLO responses. These ML approaches have revealed influential features, including the presence of diacylglycerol lipids close to the dye. On the whole, this work provides a first step toward understanding the cooperation, synergy, and interactions that occur in such complex guest-host environments, which have emerged as new targets for drug design and membrane lipid therapy.

Azide-modified corrole phosphorus complexes for endoplasmic reticulum-targeted fluorescence bioimaging and effective cancer photodynamic therapy

Study on corrole photosensitizers (PSs) for photodynamic therapy (PDT) has made remarkable progress. Targeted delivery of PSs is of great significance for enhancing therapeutic efficiency, decreasing the dosage, and reducing systemic toxicity during PDT. The development of PSs that can be specifically delivered to the subcellular organelle is still an attractive and challenging work. Herein, we synthesize a series of azide-modified corrole phosphorus and gallium complex PSs, in which phosphorus corrole 2-P could not only precisely target the endoplasmic reticulum (ER) with a Pearson correlation coefficient (PCC) up to 0.92 but also possesses the highest singlet oxygen quantum yields (ΦΔ = 0.75). This renders it remarkable PDT activity at a very low dosage (IC50 = 23 nM) towards HepG2 tumor cell line while ablating solid tumors in vivo with excellent biosecurity. Furthermore, 2-P exhibits intense red fluorescence (ΦF = 0.25), outstanding photostability, and a large Stokes shift (190 nm), making it a promising fluorescent probe for ER. This study provides a clinically potential photosensitizer for cancer photodynamic therapy and a promising ER fluorescent probe for bioimaging.

Synthesis of Highly Luminescent Silica-Coated Upconversion Nanoparticles from Lanthanide Oxides or Nitrates Using Co-Precipitation and Sol-Gel Methods.

Upconversion nanoparticles (UCNPs) are under consideration for their use as bioimaging probes with enhanced optical performance for real time follow-up under non-invasive conditions. Photostable and core-shell NaYF4:Yb3+, Er3+-SiO2 UCNPs obtained by a novel and simple co-precipitation method from lanthanide nitrates or oxides were herein synthesized for the first time. The sol-gel Stöber method followed by oven or supercritical gel drying was used to confer biocompatible surface properties to UCNPs by the formation of an ultrathin silica coating. Upconversion (UC) spectra were studied to evaluate the fluorescence of UCNPs upon red/near infrared (NIR) irradiation. ζ-potential measurements, TEM analyses, XRD patterns and long-term physicochemical stability were also assessed and confirmed that the UCNPs co-precipitation synthesis is a shape- and phase-controlling approach. The bio- and hemocompatibility of the UCNPs formulation with the highest fluorescence intensity was evaluated with murine fibroblasts and human blood, respectively, and provided excellent results that endorse the efficacy of the silica gel coating. The herein synthesized UCNPs can be regarded as efficient fluorescent probes for bioimaging purposes with the high luminescence, physicochemical stability and biocompatibility required for biomedical applications.

Synthesis, structure, photophysical property, stability of tetraphenylethylene-based boranil, and applications in cell imaging

A new family of tetraphenylethylene-based N,O-chelated boranil complexes (TPE-BAs) with aggregation-induced emission (AIE) characteristics were developed. X-ray crystallographic analysis indicated that the terminal substituents on the aniline moiety significantly affected the intermolecular stacking mode, thereby influencing the photophysical properties. The stabilities of these compounds are closely related to the substituents on the aniline moiety. Electron-donor-substituted TPE-BA-OMe exhibited the best stability, whereas the electron-acceptor-substituted compounds exhibited poor stability. Benefitting from its AIE properties and suitable lipophilicity, TPE-BA-OMe served as an excellent fluorescent probe for the specific bioimaging of lipid droplets in living cells.

Bacteria-Based Living Probes: Preparation and the Applications in Bioimaging and Diagnosis.

Bacteria can colonize a variety of in vivo biointerfaces, particularly the skin, nasal, and oral mucosa, the gastrointestinal tract, and the reproductive tract, but also target specific lesion sites, such as tumor and wound. By virtue of their prominent characteristics in motility, editability, and targeting ability, bacteria carrying imageable agents are widely developed as living probes for bioimaging and diagnosis of different diseases. This review first introduces the strategies used for preparing bacteria-based living probes, including biological engineering, chemical modification, intracellular loading, and optical manipulation. It then summarizes the recent progress of these living probes for fluorescence imaging, near-infrared imaging, ultrasonic imaging, photoacoustic imaging, magnetic resonance imaging, and positron emission tomography imaging. The biomedical applications of bacteria-based living probes are also reviewed particularly in the bioimaging and diagnosis of bacterial infections, cancers, and intestine-associated diseases. In addition, the advantages and challenges of bacteria-based living probes are discussed and future perspectives are also proposed. This review provides an updated overview of bacteria-based living probes, highlighting their great potential as a unique yet versatile platform for developing next-generation imageable agents for intelligent bioimaging, diagnosis, and even therapy.

A Neutral Flavin-Triphenylamine Probe for Mitochondrial Bioimaging under Different Microenvironments.

A bioinspired design built around a neutral flavin-triphenylamine core has been investigated for selective mitochondrial bioimaging capabilities in different microenvironments. Significant advantages with respect to long-term tracking, faster internalization, penetrability within the spheroid structures, and strong emission signal under induced hypoxia conditions have been observed, which could offer an alternative to the existing mitotrackers for hypoxia-related biological events.

Sensing Mechanism and Excited-State Dynamics of a Widely Used Intracellular Fluorescent pH Probe: pHrodo.

The pHrodo with an "off-on" response to the changes of pH has been widely used as a fluorescent pH probe for bioimaging. The fluorescence off-on mechanism is fundamentally important for its application and further development. Herein, the sensing mechanism, especially the relevant excited-state dynamics, of pHrodo is investigated by steady-state and time-resolved spectroscopy as well as quantum chemical calculations, showing that pHrodo is best understood using the bichromophore model. Its first excited state (S1) is a charge transfer state between two chromophores. From S1, pHrodo relaxes to its ground state (S0) via an ultrafast nonradiative process (∼0.5 ps), which causes its fluorescence to be "off". After protonation, S1 becomes a localized excited state, which accounts for the fluorescence being turned "on". Our work provides photophysical insight into the sensing mechanism of pHrodo and indicates the bichromophore model might be relevant to a wide range of fluorescent probes.

Advancements in bioimaging probes and nanoscale biosensors for cancer detection

Various bioimaging and biosensing tools can accurately diagnose early-stage cancer by recognizing stage-specific changes in cancer cell proliferation.

Probes for noninvasive biological visualization and biosensing of cancer cells

The early detection of tumors and precancerous conditions is vital for cancer diagnosis. Advances in fluorescence microscopic techniques and materials synthesis processes have revolutionized biomarker detection and image-guided cancer surveillance. In particular, novel materials-based diagnostic tools and innovative therapies have facilitated a precise understanding of biological processes at the molecular level. This critical review presents an overview of bioimaging probes, including functionalized chromophoric systems, non-functionalized chromophoric systems, and nanoscale biosensors. Technical challenges and future directions related to these approaches are considered.

Alendronate-derived carbon dots for bioimaging and effective osteoblastic differentiation induction and promotion

Carbon dots (CDs) belong to zero-dimensional carbon-based nanomaterials, which are promising in various biomedical applications including imaging, biosensing and drug delivery. In this work, we have synthesized novel fluorescent CDs (Alen-CDs) derived from alendronate, an anti-osteoporosis drug, through a one-step hydrothermal method. The ability of Alen-CDs for fluorescent imaging was evaluated via in vitro cellular bioimaging. In addition, the capability of Alen-CDs to induce in vitro osteoblastic differentiation was investigated by alkaline phosphatase staining, activity test and alizarin red staining. As demonstrated, Alen-CDs was effective for in vitro fluorescent imaging without obvious toxicity. Also, Alen-CDs promoted the survival of rat bone marrow mesenchymal stem cells and stimulated osteoblastic differentiation. In conclusion, Alen-CDs are a new type of nanomaterial with good biocompatibility and fluorescent properties. Alen-CDs could be applied as a potential probe for bioimaging and a promising nanodrug for bone regeneration as well as the combination to be used as a theranostic agent.

Rational Design of Tunable Near-Infrared Oxazine Probe with Large Stokes Shift for Leucine Aminopeptidase Detection and Imaging.

Near-Infrared (NIR) fluorescence imaging with the advantages of deep tissue penetration and minimum background, has been widely employed and developed in the study of biological applications. However, small Stokes shifts, difficulty in optical tuning, and pH sensitivity are still the major limitations faced by current NIR dyes. To solve these problems, we rationally designed a pH insensitive amino-tunable NIR oxazine fluorophore DQF-NH2 , which exhibited large Stokes shift (125 nm) accompanied with NIR excitation/emission due to the introduction an asymmetrical alternating vibronic feature. By benefiting from the excellent photophysical properties of DQF-NH2 , we have successfully constructed the probe DQF-NH2 -LAP with the ability to detect endogenous LAP. Bioimaging assays demonstrated that DQF-NH2 -LAP can not only effectively detect LAP in living cells, but also was successfully applied to image tumor tissue in vivo. We anticipate that the functionalizable dye DQF-NH2 may be a potential new NIR dye platform with an optically tunable group for the development of future desirable probes for bioimaging.