Fluorescent Agents Research Articles

Dual/Multi-Modal Image-Guided Diagnosis and Therapy Based on Luminogens with Aggregation-Induced Emission.

The combination of multiple imaging methods has made an indelible contribution to the diagnosis, surgical navigation, treatment, and prognostic evaluation of various diseases. Due to the unique advantages of luminogens with aggregation-induced emission (AIE), their progress has been significant in the field of organic fluorescent contrast agents. Herein, this manuscript summarizes the recent advancements in AIE molecules as contrast agents for optical image-based dual/multi-modal imaging. We particularly focus on the exceptional properties of each material and the corresponding application in the diagnosis and treatment of diseases.

Experimental In-Situ Observatory on Brownian Motion Behavior of 105 nm Sized Silica Particles During Chemical Mechanical Polishing of 4H-SiC by an Evanescent Field

The experimentally observing optical systems for on-machine measurement have been developed to study on nano-polishing phenomena during the chemical mechanical polishing process, which is a wet process in semiconductor manufacturing. The developed optical system employs an evanescent field to selectively enhance exclusively the observatory of phenomena occurring on the surface being polished, offering a lateral resolving power of approximately 400 nm, in the slurry concentration of up to 5 wt% based on the numerical aperture of the objective lens. In addition, there is also the observability of 105 nm and down to 55 nm-sized silica particles without requiring additive fluorescence agents in or around the nano-particles, even when these particles are moving on surfaces such as silica glass or hard materials (silicon carbide: 4H-SiC). Consequently, the motion behavior of nano-particles disjoining with polishing pad asperity was explored and discussed, in this paper. Experimental results revealed that the polishing pad spatially constrains the movement of particles between the pad and the substrate surface, guiding them toward the surface being polished. During pad sliding, fluidically dragged nano-particles exhibit slower movement than the polishing pad sliding speed while retaining the Brownian motion. Furthermore, 105 nm-sized silica particles did not continuously approach to attach onto the SiC surface; the nano-particles approached in steps with reduced Brownian motion in all directions before attaching. This behavior can be attributed to the effects of van der Waals attraction and electrostatic repulsion forces between the particle and the substrate surfaces.

Whole-body fluorescence cryotomography identifies a fast-acting, high-contrast, durable contrast agent for fluorescence-guided surgery.

Imaging of tumor-specific fluorescent contrast agents to guide tumor removal has been shown to improve outcomes and is now standard practice for some neurosurgical procedures. However, many agents require administration hours before surgery, a practical challenge, and may exhibit inconsistent concordance with contrast-enhanced MRI (CE-MRI), the current standard for diagnosing and guiding glioma removal. A fluorescent agent that accurately marks tumor shortly after administration and is otherwise similar to CE-MRI would help overcome these shortcomings. Methods: We used whole-body 3-D fluorescence cryo-imaging and co-registered CE-MRI volumes to evaluate several fluorescent contrast agent candidates for diagnostic performance and concordance with CE-MRI. Mice with brain tumors were administered a cocktail of fluorescent agent candidates and a MRI contrast agent, and then imaged with MRI and fluorescence cryo-imaging at several timepoints after administration. The high-resolution 3-D cryo-imaging volumes of the fluorescent agents were used to determine diagnostic performance metrics and correlation with CE-MRI. Results: While all agents showed positive metrics, one agent, tetramethylrhodamine conjugated to a small polyethylene glycol chain (TMR-PEG1k), outperformed the others, exhibiting minimal normal brain signal, high tumor-to-background-ratio, diagnostic accuracy, and cross-correlation to CE-MRI at all post-administration timepoints (10-90 min) and tumor lines examined. Conclusion: These favorable properties establish TMR-PEG1k as a promising candidate for surgical guidance.

Near-IR nanolignin sensitizers based on pyrene conjugated chlorin and bacteriochlorin for ROS generation, DNA Intercalation and bioimaging

Near-infrared (NIR) fluorescent agents are extensively used for biomedical imaging due to their ability towards deep tissue penetration. Tetrapyrrole-based photosensitizers are promising candidates in this regard. Further, extended conjugation of...

Efficient one-step amide formation using amino porphyrins.

Amide bonds are one of the most prevalent phenomena in nature and are utilized frequently in drug and material design. However, forming amide bonds is not always efficient or high yielding, particularly when the amine used to conjugate to a carboxylic acid is a weak nucleophile. This limitation precludes many useful amino compounds from participating in conjugation reactions to form amides. A particularly valuable amino compound, which is also a very weak nucleophile, is the amino porphyrin, valued for its role as a photosensitizer, fluorescent agent, catalyst, or, upon metalation, even a very efficient contrast agent for magnetic resonance imaging (MRI). In this work, we propose fast and high-yield coupling of an unreactive amine - the amino porphyrin - to carboxylic acid via isothiocyanate conjugation. Reactions can be achieved in one step at room temperature in one hour, achieving quantitative conversion and near perfect selectivity. Both metalated and unmetalated porphyrin, as well as fluorescein isothiocyanate (FITC), demonstrated efficient conjugation. To illustrate the value of the proposed method, we created a new blood-pool MRI contrast agent that reversibly binds to serum albumin. This new blood-pool agent, known as MITC-Deox (MRI isothiocyanate that links with deoxycholic acid), substantially reduced T1 relaxation times in blood vessels in mice, remained stable for 1 hour, cleared from blood by 24 hours, and was eliminated from the body after 4 days. The proposed method for efficient amide formation is a superior alternative to existing coupling methods, opening a door to novel synthesis of MRI contrast agents and beyond.

Progress of fluorescence imaging in lymph node dissection surgery for prostate and bladder cancer.

Fluorescence imaging is a relatively new imaging method used to visualize different tissue structures to help guide intraoperative operations, which has potential advantages with high sensitivity and contrast compared to conventional imaging. In this work, we review fluorescent contrast agents and devices used for lymphatic system imaging. Indocyanine green is the most widely utilized due to its high sensitivity, specificity, low background fluorescence, and safety profile. In prostate and bladder cancer lymph node dissection, the complex lymphatic drainage can result in missed metastatic nodes and extensive dissection increases the risk of complications like lymphocele, presenting a significant challenge for urologists. Fluorescence-guided sentinel lymph node dissection facilitates precise tumor staging. The combination of fluorescence and radiographic imaging improves the accuracy of lymph node staging. Multimodal imaging presents new potential for precisely identifying metastatic pelvic lymph nodes.

Biocompatible and low-cost iodine-doped carbon dots as a bifunctional fluorescent and radiocontrast agent for X-ray CT imaging

Biocompatible iodine-doped carbon dots exhibiting high X-ray attenuation efficiency and fluorescent properties have been produced using low-cost reagents.

A radioactive and fluorescent dual modality cysteine cathepsin-B activity-based probe for the detection and treatment evaluation in rheumatoid arthritis.

Activated macrophages are key effector cells and specific markers in patients with rheumatoid arthritis (RA). Cysteine cathepsin B (CTS-B) is highly expressed in macrophages and positively associated with RA activity and severity. This study aims to evaluate an activity-based multi-modality diagnostic agent, 68Ga-BMX2, which targets CTS-B to visualize the arthritis activity and evaluate the treatment efficacy. A CTS-B activity-based probe, BMX2, was labeled efficiently with 68Ga to produce 68Ga-BMX2 for fluorescent and positron emission tomography (PET) multi-modality imaging. The affinity and specificity of BMX2 binding with the CTS-B enzyme in macrophages were determined by radioactive experiment using RAW 264.7 cell lines, with CA074 and BMX5 as the inhibitors to test the specificity of the binding. Then, PET and fluorescence imaging were acquired on collagen-induced arthritis (CIA) mice. Additionally, the treatment monitoring capability of 68Ga-BMX2 PET/CT imaging was tested with methotrexate (MTX). RAW 264.7 macrophage cells showed significant uptake of 68Ga-BMX2. The binding of BMX2 with CTS-B in RAW 264.7 macrophage cells is time-dependent and could be blocked by CA074 and BMX5. In vivo optical and PET imaging showed high signals in the right hind arthritis in CIA mice from 68Ga-BMX2 and BMX2 accumulated for at least 120 h. Additionally, 68Ga-BMX2 signals were significantly reduced in the MTX-treated CIA mice compared to the control group. The 68Ga-BMX2, a radioactive and fluorescent dual-modality diagnostic agent targeting CTS-B, demonstrated a practical approach for CIA PET and fluorescence imaging. The 68Ga-BMX2 multimodality imaging could significantly monitor the treatment response in the CIA mice.

A highly fluorescent and readily accessible all-organic photosensitizer model for advancing image-guided cancer PDT.

The potential of using image-guided photodynamic therapy (ig-PDT) for cancer, especially with highly biocompatible fluorescent agents free of heavy atoms, is well recognized. This is due to key advantages related to minimizing adverse side effects associated with standard cancer chemotherapy. However, this theragnostic approach is strongly limited by the lack of synthetically-accessible and easily-modulable chemical scaffolds, enabling the rapid design and construction of advanced agents for clinical ig-PDT. In fact, there are still very few ig-PDT agents clinically approved. Herein we report a readily accessible, easy-tunable and highly fluorescent all-organic small photosensitizer, as a model design for accelerating the development and translation of advanced ig-PDT agents for cancer. This scaffold is based on BODIPY, which assures high fluorescence, accessibility, and ease of performance adaptation by workable chemistry. The optimal PDT performance of this BODIPY dye, tested in highly resistant pancreatic cancer cells, despite its high fluorescent behavior, maintained even after fixation and cancer cell death, is based on its selective accumulation in mitochondria. This induces apoptosis upon illumination, as evidenced by proteomic studies and flow cytometry. All these characteristics make the reported BODIPY-based fluorescent photosensitizer a valuable model for the rapid development of ig-PDT agents for clinical use.

A novel BODIPY-based theranostic agent for in vivo fluorescence imaging of cerebral Aβ and ameliorating Aβ-associated disorders in Alzheimer's disease transgenic mice.

β-Amyloid (Aβ) aggregation is increasingly recognized as both a biomarker and an inducer of the progression of Alzheimer's disease (AD). Here, we describe a novel fluorescent probe P14, developed based on the BODIPY structure, capable of simultaneous visualization and inhibition of Aβ aggregation in vivo. P14 shows high binding affinity to Aβ aggregates and selectively labels Aβ plaques in the brain slices of APP/PS1 mice. Moreover, P14 is able to visualize overloaded Aβ in both APP/PS1 and 5 × FAD transgenic mice in vivo. From the aspect of potential therapeutic effects, P14 administration inhibits Aβ aggregation and alleviates Aβ-induced neuronal damage in vitro, as well as reduces central Aβ deposition and ameliorates cognitive impairment in APP/PS1 transgenic mice in vivo. Finally, P14 is applied to monitor the progression of Aβ aggregation in the brain of 5 × FAD transgenic mice and the intervention effect itself by fluorescence imaging. In summary, the discovery of this fluorescent agent might provide important clues for the future development of theranostic drug candidates targeting Aβ aggregation in AD.

Single-molecule detection of a terrylenediimide-based near-infrared emitter.

Environment-sensitive fluorescent agents with near-infrared (NIR) emission are in great demand owing to their applications in biomedical and quantum technologies. We report a novel NIR absorbing (λ Abs max = 734 nm) and emitting (λ Fl max = 814 nm) terrylenediimide (TDI) based donor-acceptor chromophore (TDI-TPA4), exhibiting polarity-sensitive single-photon emission. By virtue of the charge transfer (CT) character, ensemble level measurements revealed solvatochromism and NIR emission (ϕ Fl = 26.2%), overcoming the energy gap law. The CT nature of the excited states is further validated by state-of-the-art fragment-based excited state theoretical analysis. To mimic the polarity conditions at the single-molecule level, TDI-TPA4 was immobilized in polystyrene (PS; low polar) and poly(vinyl alcohol) (PVA; high polar) matrices, which enables tuning of the energy levels of the locally excited state and charge-separated (CS) state. Minimal blinking and prolonged survival time of the TDI-TPA4 molecule in the PS matrix, in contrast to the PVA matrix, possibly confirms the implication of the energy gap law and polarity sensitivity of TDI-TPA4. The existence of the CT state in nonpolar and CS state in polar solvents was confirmed by transient absorption measurements in the femtosecond regime. The current work sheds light on the design principle for NIR single-photon emitting organic chromophores for deep tissue imaging and probing the nanoscale heterogeneity.

Self-healing coating with the ability to mark damaged locations, based on electrospun photothermal fibers and AIE agents

The anti-corrosion coatings based on core-sheath fibers prepared by electrospinning methods are developed for the indication and repair of large-scale damage to coatings. The cores are the composite fluorescent agents composed of AIE-gens and CHEF-gens, which exhibit strong fluorescence characteristics sensitive to corrosion due to the CHEF and FRET effects. The composite fluorescent agents provided information about the location of damages when the coatings, with the fibers as the core materials, are damaged. The sheaths are photothermal materials, composed of photothermal responsive acid doped polyaniline and shape memory polyurethane. The photothermal properties of acid doped polyaniline, under the irradiation of an infrared laser, contributes to heating in the damaged areas. The softened and activated polyurethane simultaneously lead to interpenetration and entanglement of molecular chains at the closed damage sites, resulting in complete repair of the damaged coatings.

In vivo visualization of tumor-associated macrophages re-education by photoacoustic/fluorescence dual-modal imaging with a metal-organic frames-based caspase-1 nanoreporter

Tumor-associated macrophages (TAMs) are vital in the tumor microenvironment, contributing to immunosuppression and therapy tolerance. Despite their importance, the precise re-education of TAMs in vivo continues to present a formidable challenge. Moreover, the lack of real-time and efficient methods to comprehend the spatiotemporal kinetics of TAMs repolarization remains a significant hurdle, severely hampering the accurate assessment of treatment efficacy and prognosis. Herein, we designed a metal–organic frameworks (MOFs) based Caspase-1 nanoreporter (MCNR) that can deliver a TLR7/8 agonist to the TAMs and track time-sensitive Caspase-1 activity as a direct method to monitor the initiation of immune reprogramming. This nanosystem exhibits excellent TAMs targeting ability, enhanced tumor accumulation, and stimuli-responsive behavior. By inducing the reprogramming of TAMs, they were able to enhance T-cell infiltration in tumor tissue, resulting in inhibited tumor growth and improved survival in mice model. Moreover, MCNR also serves as an activatable photoacoustic and fluorescent dual-mode imaging agent through Caspase-1-mediated specific enzyme digestion. This feature enables non-invasive and real-time antitumor immune activation monitoring. Overall, our findings indicate that MCNR has the potential to be a valuable tool for tumor immune microenvironment remodeling and noninvasive quantitative detection and real-time monitoring of TAMs repolarization to immunotherapy in the early stage.

Efficient adsorption of fluorescent whitening agent 351 from high salinity environmental water by polymer supported hydroxyl functional ionic liquid

Fluorescent whitening agent 351 (FWA351) is an emerging highly persistent pollutant with potentially harmful effects on human health and aquatic ecosystem. However, developing highly effective adsorbents for the pre-treatment of environmental water samples containing FWA351 remains a problem. In this study, a novel polymer supported hydroxyl-functionalized ionic liquid (PS-[HMMim][Cl]) was prepared via a simple one-step method and used for the removal of FWA351 from water. It was found that the PS-[HMMim][Cl] exhibited an ultrahigh adsorption capacity of 1203.6 mg/g for FWA351 and maintained high removal efficiency in a wide pH range (2.0–9.0). Notably, even the concentration of FWA351 was as low as 0.005 mg/L (5 ppb), FWA351 could be effectively removed by the adsorbent, achieving an enrichment factor as high as 447.8. Various inorganic salts (such as KCl, NaCl and MgCl2) at the concentration up to 1.0 mol/L had no obvious effect on the removal efficiency. The PS-[HMMim][Cl] also displayed outstanding adsorption performance towards fluorescent whitening agents (FWAs) with similar structure to FWA351 and could be regenerated and reused for at least 8 cycles. Finally, the adsorbent was applied for the pre-treatment and then detection of FWA351 in six kinds of environmental water samples. Mechanism studies suggest that the synergistic driving forces of electrostatic attraction, hydrogen bonding, π-π interaction and ion exchange are responsible for the efficient adsorption of FWA351. This work provides a new strategy for efficient adsorption and analysis of FWA351.

The Interplay of Protein Hydrolysis and Ammonia in the Stability of Hevea Rubber Latex during Storage.

Natural rubber (NR) latex derived from Hevea brasiliensis is a complex colloid comprising mainly rubber hydrocarbons (latex particles) and a multitude of minor non-rubber constituents such as non-rubber particles, proteins, lipids, carbohydrates, and soluble organic and inorganic substances. NR latex is susceptible to enzymatic attack after it leaves the trees. It is usually preserved with ammonia and, to a lesser extent, with other preservatives to enhance its colloidal stability during storage. Despite numerous studies in the literature on the influence of rubber proteins on NR latex stability, issues regarding the effect of protein hydrolysis in the presence of ammonia on latex stability during storage are still far from resolved. The present work aims to elucidate the interplay between protein hydrolysis and ammoniation in NR latex stability. Both high- and low-ammonia (with a secondary preservative) NR latexes were used to monitor the changes in their protein compositions during storage. High-ammonia (FNR-A) latex preserved with 0.6% (v/v) ammonia, a low 0.1% ammonia/TMTD/ZnO (FNR-TZ) latex, and a deproteinized NR (PDNR) latex were labeled with fluorescence agents and observed using confocal laser scanning microscopy to determine their protein composition. Protein hydrolysis was confirmed via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results revealed that protein hydrolysis increased with the storage duration. The change in protein composition accompanying hydrolysis also allows the spatial distribution of allergenic proteins to be estimated in the latex. Concurrently, the latex stability increased with the storage duration, as measured by the latex's mechanical stability time (MST) and the zeta potential of the latex particles. As monitored by AFM, the surface roughness of the NR latex film increased markedly during extended storage compared with that of the DPNR latex, which remained smooth. These results underscore the pivotal role of ammonia in bolstering NR latex stability brought on by protein hydrolysis, which greatly impacts latex film's formation behavior. NR latex stability underpins the quality of latex-dipped goods during manufacturing, particularly those for medical gloves.

Fluorescence-guided pediatric surgery: The past, present, and future

Fluorescence-guided surgery (FGS) is becoming a common practice as an intraoperative aid in both adult and pediatric populations. It makes use of fluorescent contrast agents to identify targeted structures intraoperatively for precise resection with the help of a near-infrared (NIR) light camera. A wide spectrum of contrast agents and imaging platforms has been developed throughout the years to enhance their use for intraoperative tissue differentiation. FGS is applied in pediatric surgery for various indications, including thoracic, gastrointestinal, hepatobiliary, urogenital, oncology, and vascular surgery. Despite the popularity of fluorescence-guided pediatric surgery, the current literature is limited. The present review offers an up-to-date appraisal of the development and current status of FGS in the field of pediatric surgery.

An In Vivo Dual-Observation Method to Monitor Tumor Mass and Tumor-Surface Blood Vessels for Developing Anti-Angiogenesis Agents against Submillimeter Tumors.

Managing metastasis at the early stage and detecting and treating submillimeter tumors at early metastasis are crucial for improving cancer prognosis. Angiogenesis is a critical target for developing drugs to detect and inhibit submillimeter tumor growth; however, drug development remains challenging because there are no suitable models for observing the submillimeter tumor mass and the surrounding blood vessels in vivo. We have established a xenograft subcutaneous submillimeter tumor mouse model with HT-29-RFP by transplanting a single spheroid grown on radiation-crosslinked gelatin hydrogel microwells. Here, we developed an in vivo dual-observation method to observe the submillimeter tumor mass and tumor-surface blood vessels using this model. RFP was detected to observe the tumor mass, and a fluorescent angiography agent FITC-dextran was administered to observe blood vessels via stereoscopic fluorescence microscopy. The anti-angiogenesis agent regorafenib was used to confirm the usefulness of this method. This method effectively detected the submillimeter tumor mass and tumor-surface blood vessels in vivo. Regorafenib treatment revealed tumor growth inhibition and angiogenesis downregulation with reduced vascular extremities, segments, and meshes. Further, we confirmed that tumor-surface blood vessel areas monitored using in vivo dual-observation correlated with intratumoral blood vessel areas observed via fluorescence microscopy with frozen sections. In conclusion, this method would be useful in developing anti-angiogenesis agents against submillimeter tumors.

Robot-assisted navigation for percutaneous localization of peripheral pulmonary nodule: an in vivo swine study.

Robot-assisted surgery (RAS) systems have been developed but rarely applied to lung nodule localization. This study aimed to assess the feasibility and safety of using a robot-assisted navigation system in percutaneous lung nodule localization. A computed tomography (CT)-guided robot-assisted navigation system was used to localize the simulated peripheral nodule in the swine lung through fluorescent agent injection. After the localization, fluorescent thoracoscopic wedge resection was performed. The deviation between the target point and the needle tip was measured using a professional 3-dimensional (3D) distance measurement software. The primary outcome was the localization accuracy (deviation) of the localization. The secondary outcomes were the localization-related complication rate, the localization duration, and the success rate. A total of 4 pigs were enrolled, and 20 peripheral lung nodules were created and localized successfully. All nodules underwent subsequent wedge resection for verification. The mean deviation by measuring the 3D distance was 3.81 mm [standard deviation (SD): 1.29 mm, 95% confidence interval (CI): 2.936-4.536 mm]. The technical success rate for localization was 100%, and the mean localization time was 14.69 minutes (SD: 4.67 minutes). The complication rate was 5% (1/20), with 1 pneumothorax after localization, and no mortality occurred. This pilot animal study demonstrated the promising potential of the robot-assisted navigation technique in peripheral lung nodule localization, with high accuracy and feasibility. Further clinical trials are needed to validate its safety compared to traditional manual localization.

Design, synthesis, cell imaging, and bioactivity assessment of novel Rhodamine-Pyrimidine nido-carborane derivatives as fluorescent anticancer agents

Because of the advantages of fluorescent drugs consisting of fluorescent dyes and compounds with anticancer activity for therapeutic diagnostics and personalized medicine. A series of fluorescent antitumor drugs rhodamine pyrimidine derivatives (RP1, RP2, RP3 and RP4) were synthesized by linking rhodamine B and pyrimidine derivatives through ethylenediamine bridge and then introduced into nido-carborane to form fluorescent salt derivatives (RPB1, RPB2, RPB3, and RPB4). It improves the tumor targeting and imaging ability of pyrimidine derivatives. By CCK8, it was found that the inhibition rates of RP4 and RPB4 at 10 μg/mL on HeLa, PC-3 and L02 cells were 70.4 %, 68.7 %, 86.1 % and 72.5 %, 76.4 %, 81.0 %, respectively. The compound has strong inhibitory effect on cancer cells. The logP values of compounds RP4 and RPB4 were between 0 and 3, which were easily absorbed and transported through the cell membrane. Further cell imaging experiments showed that RP4 could enter tumor cells through cell membrane and had good biocompatibility. The experimental results show that compounds RP4 has good imaging capability and tumor targeting, and is a potential fluorescent anticancer agents.

Metal Dipicolylamines and their Biomedical Applications: A Mini Review

Research in synthetic inorganic chemistry demonstrates that metal complexes are widely utilized as therapeutic as well as diagnostic agents. Due to the coordinative saturation, substitutional inertness, and unique redox characteristics, metal polypyridyl complexes have ignited attention and have been exploited in a number of biological and biomedical fields. The polypyridyl ligand, dipicolylamine (DPA) is a symmetrical secondary amine with two pyridyl rings. Delocalization of excited electrons occurs throughout the ligand system due to its conjugated character and the metal-to-ligand charge transfer transitions, resulting in a strong fluorescence signal. The high lipophilic nature of this ligand has proven to improve metal-DPA complex absorption by cell membranes. The N–H amine group in DPA and its analogues have the additional function of allowing hydrogen bonds to form, and deprotonation of this amine has allowed the creation of mononuclear and polynuclear species, with deprotonation of the bridging amine nitrogen also playing a significant role in the coordination sequence. DPA-appended metal complexes have garnered attention in the quest for linkages between magnetic, spectroscopic, structural, and coordination geometries. A number of mononuclear metal-DPA complex crystal structures with four, five, or six coordinated metal centers have been synthesized and explored for their biological properties. This study reviews DPA-derivatized ligand-linked metal complexes as promising cancer, microbial, and fungal inflammatory therapeutics, as well as potential diagnostic agents for fluorescence imaging and radiopharmaceuticals.
 Keywords: Dipicolylamine, metal complexes, coordination geometries, fluorescent agents