Fluorescent Agents Research Articles

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

A new bimodal approach for sentinel lymph node imaging in prostate cancer using a magnetic and fluorescent hybrid tracer

PurposeTo obtain initial data on sentinel lymph node (SLN) visualisation by pre-operative magnetic resonance imaging (MRI) and intra-operative bimodal SLN identification using a new magnetic fluorescent hybrid tracer in prostate cancer (PCa) patients.MethodsTen patients at > 5% risk for lymph node (LN) invasion were included. The day before surgery, a magnetic fluorescent hybrid tracer consisting of superparamagnetic iron oxide nanoparticles (SPION) and indocyanine green was transrectally injected into the prostate. Five hours after injection, transversal pelvic MRI scans were recorded and T2*-weighed images were screened for pelvic LNs with SPION uptake. Intra-operatively, magnetically active and/or fluorescent SLNs were detected by a handheld magnetometer and near-infrared fluorescence imaging (FI). Extended pelvic lymph node dissection (PLND) and radical prostatectomy completed the surgery. All resected specimens were checked ex situ for magnetic activity and fluorescence and were histopathologically examined.ResultsPre-operative MRI identified 145 pelvic LNs with SPION uptake. In total, 75 (median 6, range 3‒13) magnetically active SLNs were resected, including 14 SLNs not seen on MRI. FI identified 89 fluorescent LNs (median 8.5, range 4‒13) of which 15 LNs were not magnetically active. Concordance of the different techniques was 70% for pre-operative MRI vs. magnetometer-guided PLND and 88% for magnetic vs. fluorescent SLN detection.ConclusionThese are the first promising results of bimodal, magnetic fluorescent SLN detection in PCa patients. Our magnetic fluorescent hybrid approach provides the surgeon a pre-operative lymphatic roadmap by using MRI and intra-operative visual guidance through the application of a fluorescent lymphatic agent. The diagnostic accuracy of our new hybrid approach has to be evaluated in further studies.Trial registrationDRKS00032808. Registered 04 October 2023, retrospectively registered.

Enhanced tumor targeting with near-infrared light-activated indocyanine green encapsulated in covalent organic framework for combined photodynamic therapy (PDT) and photothermal therapy (PTT)

Covalent organic framework (COF), a new emerging class of crystalline porous polymeric networks with reversible covalent bonds are a better smart material in Nano medicine in this regard. 1,3,5-Triformylbenzene (TFB) and 2,5-Bis(methylsulfonyl)-1,4-diaminobenzene (BMSDAB) are used to develop a uniform shaped covalent organic framework at room temperature (RT-USCOF). In order to achieve the PTT and PDT effect in RT-USCOF, near infrared photosensitizer, photothermal and fluorescence agent, indocyanine green (ICG) was encapsulated. Moreover, enhanced loading content of ICG in I RT-USCOF to produce II RT-USCOF followed by stabilize with hyaluronic acid designed in order to reduce drug leakage and increase carrier stability. The II RT-USCOF-HA exhibits an increased encapsulation efficiency of 57.75% and a drug loading capacity of 53.59%. The developed system shows an 80% ICG release upon exposure to NIR 808-laser in acidic pH relative to the one without light irradiation proved the effective structural changes under light cause drug release. The newly developed II RT-USCOF-HA shows excellent biocompatibility against NIH 3T3 cell lines and CT2A cell lines under dark. Whereas, upon irradiation with NIR 808 nm laser, the system kills more than 70% of the cells and cellular uptake studies also proved the same. Finally, flow cytometry analysis confirmed, the II-RT-USCOF-HA treated with CT2A cell lines shows an early apoptosis cell population of 1.95%, a late apoptosis rate of 1.24% and a necrotic cell death of 4.77%, which are significantly greater than the free ICG treated groups. The singlet oxygen production followed by thermal ablation of tumor cells by the developed system further demonstrates its efficacy as a PDT/PTT probe.

De Novo Design of Near-Infrared Fluorescent Agents Activated by Peroxynitrite and Glutathione-Responsive Imaging for Diabetic Liver Disease.

Diabetes and its complications, such as diabetes liver disease, is a major problem puzzling people's health. The detection of redox states in its pathological process can effectively help us gain a deeper understanding of the disease. The pair of oxidation-reduction substances peroxynitrite (ONOO- ) and glutathione (GSH) is considered to be closely related to their occurrence and development. Thus, direct visualization of ONOO- and GSH in diabetes liver disease is critical to evaluate the disease at the molecular level. Herein, two activatable agents NTCF-ONOO- and NTCF-GSH are prepared for selectively detecting ONOO- and GSH through protection and deprotection strategies based on hydroxyl and amino groups of near-infrared fluorophore. Fluorescence imaging of exogenous and endogenous ONOO- and GSH changes in living cells and in vivo is observed. The ONOO- and GSH level in the diabetes liver disease cellular model are visualized and the possible redox imbalance mechanism related to the oxidized (NAD+ ) and reduced (NADH) nicotinamide adenine dinucleotides is explored in this process. Moreover, these probes can sensitively recognize ONOO- and GSH in the process of oxidative stress resulting from streptozotocin and streptozotocin/acetaminophen-induced complex diabetic liver disease in vivo. In addition, they can be applied for monitoring the clinical serum sample related with diabetic patients.

Flushing effect of micro-bolus pulse injection and its potential impact on peripheral veins under hemostasis.

To explore the feasibility of using micro-bolus pulse injection method to reduce the dilution effect of pipeline on high concentration injection, and to understand low liquid volume bolus injection based on low injection speed. Using a programmable pulse injection pump, a 25-cm - long pipeline containing water-soluble fluorescent agent was flushed using different volumes of bolus, and the time spent for the complete disappearance of the fluorescent agent was recorded to evaluate the flushing efficiency. The finite element simulation of 2-phase flow was carried out using computational fluid dynamics (CFD) technology, and the difference of shear rate and pressure distribution in the vein of pulse injection and direct injection of bolus under hemostasis was compared and simulated. Micro-bolus pulse flushing has advantages in completing perfusion imaging applications, such as small volume imaging agent injection. Compared with non-pulse injection, the effective flushing volume decreases by 49.7%, the average injection speed decreases by 56%, and the maintenance time of high shear rate is shorter when using micro-bolus pulse injection. The impact of micro-bolus pulse injection on the vein can achieve the same or even lower negative effects as other injection methods after increasing the hemostatic distance to 100 mm. In the case of bolus injection requiring high concentration and small volume, such as for radiopharmaceutical dynamic imaging, the application of micro-bolus pulse injection is an effective way to overcome the dilution phenomenon of the imaging agent in the pipeline. During hemostasis, the micro-bolus pulse injection needs to control a longer hemostasis distance to reduce the potential impact on peripheral veins.

Design of a serum albumin sensitive probe for cell imaging and drug delivery by modifying a fluorescent agent

Protein-ligand interaction serves as a critical foundation for intramolecular charge transfer (ICT) probes to bind and recognize proteins. Many fluorescent agents are reported to have favorable protein binding abilities but lack protein sensitivity. Herein, a new serum albumin sensitive ICT probe (named MBD) was synthesized by modifying an existing fluorescent agent MPM. MBD not only inherited the favorable human serum albumin (HSA) binding ability of MPM but also newly obtained favorable serum albumin responsiveness. These properties were maintained after MBD formed a complex with water-soluble pillar[5]arene (WP5). Typical commercial drugs, fluorescent amino acids, fatty acids, and metal ions had negligible effects on the HSA responsiveness of the complex. The WP5-MBD complex possessed strong anti-interference ability in binding and recognizing HSA. Moreover, the WP5-MBD complex self-assembled into supramolecular fluorescent nanoparticles in solution. Such nanoparticles possessed good cell imaging and traceable drug delivery properties, which hold great potential for cancer treatment and diagnosis.

Fluorescent Conversion Agent Embedded in Zinc Oxide as an Electron-Transporting Layer for High-Performance Non-Fullerene Organic Solar Cells with Improved Photostability.

Zinc oxide (ZnO) is widely used as an electron transporting layer (ETL) for organic solar cells (OSCs). Here, a low-cost commercial water/alcohol-soluble fluorescent conversion agent, sodium 2,2'-([1,1'-biphenyl]-4,4'-diyldivinylene)-bis(benzenesulfonate) (CBS), is incorporated into ZnO to develop a novel organic-inorganic hybrid ETL for high-performance OSCs. The photoinduced charge transfer from CBS to ZnO significantly improves the charge transport properties of ZnO, resulting in faster electron extraction and reduced charge recombination in OSC devices with ZnO:CBS ETLs. ZnO:CBS-based devices exhibit higher power conversion efficiencies (PCEs) than their pure ZnO-based counterparts, especially in devices with a thicker ETL, which is more suitable for roll-to-roll and large-area module processing. Furthermore, the strong ultraviolet-light absorption capability of CBS inhibits the photodegradation of the active layer, improving the photostability of ZnO:CBS based OSC devices. Therefore, this work provides a simple and effective strategy for realizing high-performance OSCs with high PCE and good photostability, which can further facilitate the commercialization of OSCs.

Near‐Infrared Emission‐Cyanine 5.5 and Doxorubicin Loaded Magnetite Nanoparticles for Dual In Vivo Magnetic Resonance and Biodistribution Imaging

AbstractThe in vivo imaging of the drug distribution during cancer treatment plays an important role in helping physicians and pharmacists to monitor the progress of treatment and improve the effectiveness of therapy. Our goal in this work was to prepare a multifunctional nano drug delivery system that not only co‐loaded iron oxide nanoparticles (Fe3O4 NPs) and Doxorubicin (Dox) but also attached the near‐infrared fluorescent agent Cyanine 5.5 based on poly(lactide)‐tocopheryl polyethylene glycol 1000 succinate (PLA‐TPGS) copolymer. Cyanine 5.5 was attached to the surface Fe3O4 NPs via the NH−CO chemical bond, followed by co‐loading of Dox onto the PLA‐TPGS copolymer by the emulsion solvent evaporation method. The structural, morphological, dimensional, stability, and optical‐magnetic properties of the as‐synthesized nanosystem were fully characterized. From the drug‐released study, the prolonged release of Dox accelerated with increasing acidic conditions (at pH 5.0 and 6.5), while a slow release was observed at pH 7.4. Through the MTT assay, the nanosystem had cellular toxicity against Hep−G2 and HeLa cell lines with low IC50 values of 0.42 and 0.78 μg.mL−1 in terms of Dox concentration, respectively. At a magnetic field of 7 T, this nanosystem also exhibited a high r2/r1 ratio of 164.5 and a strong contrast magnetic resonance (MR) image in the liver. In vivo biodistribution studies revealed that the highest fluorescence intensity achived in the liver after 6 h, and the nanosystem was eliminated from the body after at least 24 h. Collectively, these results demonstrated that the PLA‐TPGS‐Fe3O4‐Cyanine 5.5‐Dox nanosystem is a good candidate for simultaneously increasing contrast for MR and near‐infrared fluorescence imaging.

Multifunctional nucleoside-AIEgens bearing quaternary ammonium cationic for reversible response, bioimaging, and antibacterial

A multifunctional nucleoside-based AIEgens sensor (TPEPy-dU) was constructed for visual screening of Hg2+, determine to the reversible response of Fe3+ and biothiols, and applied for cell imaging, and drug-free bacterial killing. The TPEPy-dU displayed 10-folds fluorescence enhancement at 540 nm of emission in response to trace Hg2+ ions with 10 nM of LOD, which can be immediately quenched by adding Fe3+ or GSH/Cys-containing sulfhydryl groups. Moreover, their bacterial staining efficiency closely correlates with their antibacterial efficacy as they demonstrated comparatively higher antibacterial activity against Gram-positive bacteria than Gram-negative bacteria. The drug-free antibacterial results involved the stating prominent surface damages at the sites of interactions between bacterial cells and TPEPy-dU that were further verified by CLSM and SEM images. It can be applied as a potential fluorescent agent to explore the related antibacterial mechanisms for treating and monitoring bacterial infections in vivo due to their nontoxic nature. Compared with conventional sensors and antibacterial therapies, these findings elevated the synthetic strategies of fluorescent probes and represented an advanced antibacterial agent wearing quaternary ammonium cationic with low resistance in clinical diagnosis.

An ultrasensitive flexible strain sensor based on CNC/CNTs/MXene/TPU fibrous mat for human motion, sound and visually personalized rehabilitation training monitoring

Personalized rehabilitation training provides maximum help to stroke patients to alleviate the after-effects and restore the body to normal function. However, available monitoring devices have the disadvantages of being large, requiring professional guidance, and lacking intuitive signal display capabilities. Herein, a bio-inspired wearable high-performance strain sensor with a simple structure can simultaneous electrical signals and optical visualization in response to external stimuli. The sensor comprises a conductive layer with significant electromechanical behaviors of cellulose nanocrystals (CNC)/carbon nanotubes (CNTs)/MXene nanohybrid network, and a stretchable elastomer layer consisting of thermoplastic polyurethane and fluorescent agent. Benefiting from the designed microcracks and fluorescent material, the strain sensor exhibits ultra-high sensitivity (476800), ultra-low detection limit (0.005%), low response time (60 ms), wide working range (0–100%), and enables strain visualization for applications in visually rehabilitation training monitoring. Based on these sensing characteristics, the sensor shows great advantages in human motion and sound monitoring, with the integration of digital signals and visual images makes it show great potential in visually personalized rehabilitation training monitoring.

Next-generation agents for fluorescence-guided glioblastoma surgery.

Glioblastoma is a fast-growing and aggressive form of brain cancer. Even with maximal treatment, patients show a low median survival and are often subjected to a high recurrence incidence. The currently available treatments require multimodal management, including maximal safe surgical resection, followed by radiation and chemotherapy. Because of the infiltrative glioblastoma nature, intraoperative differentiation of cancer tissue from normal brain parenchyma is very challenging, and this accounts for the low rate of complete tumor resection. For these reasons, clinicians have increasingly used various intraoperative adjuncts to improve surgical results, such as fluorescent agents. However, most of the existing fluorophores show several limitations such as poor selectivity, photostability, photosensitization and high costs. This could limit their application to successfully improve glioblastoma resection. In the present perspective, we highlight the possibility to develop next-generation fluorescent tools able to more selectively label cancer cells during surgical resection.

Fluorescent whiteness measurement of textiles by multispectral imaging system

AbstractAccurate whiteness measurement is critical across various industries including textiles, paper, and detergents. With the advent of fluorescent whitening agents (FWAs), the assessment of whiteness has evolved from simply measuring reflectance to determining the spectral radiance factor (SRF) of materials. While multispectral imaging (MSI) can effectively measure object reflectance and colour, its accuracy is compromised by FWAs that significantly augment spectral reflectance. Thus, reliable whiteness measurement requires the determination of both reflectance and SRF. This article presents the design of a specialised light source equipped with an ultraviolet (UV) filter to provide adjustable illumination within an integrating sphere system. By tuning the position of the UV filter, multispectral images are captured of a fabric under varying UV exposures. These images are subsequently processed and combined to reconstruct full‐range visible spectrum information. This approach is shown to achieve high accuracy and spatial uniformity in quantifying the whiteness of textiles using MSI.

A commentary on 'Approved and investigational fluorescent optical imaging agents for disease detection in surgery'.

A commentary on 'Approved and investigational fluorescent optical imaging agents for disease detection in surgery'.

Fast ICCD-based temperature modulated fluorescence tomography.

Fluorescence tomography (FT) has become a powerful preclinical imaging modality with a great potential for several clinical applications. Although it has superior sensitivity and utilizes low-cost instrumentation, the highly scattering nature of bio-tissue makes FT in thick samples challenging, resulting in poor resolution and low quantitative accuracy. To overcome the limitations of FT, we previously introduced a novel method, termed temperature modulated fluorescence tomography (TMFT), which is based on two key elements: (1)temperature-sensitive fluorescent agents (ThermoDots) and (2)high-intensity focused ultrasound (HIFU). The fluorescence emission of ThermoDots increases up to hundredfold with only several degree temperature elevation. The exceptional and reversible response of these ThermoDots enables their modulation, which effectively allows their localization using the HIFU. Their localization is then used as functional a priori during the FT image reconstruction process to resolve their distribution with higher spatial resolution. The last version of the TMFT system was based on a cooled CCD camera utilizing a step-and-shoot mode, which necessitated long total imaging time only for a small selected region of interest (ROI). In this paper, we present the latest version of our TMFT technology, which uses a much faster continuous HIFU scanning mode based on an intensified CCD (ICCD) camera. This new, to the best of our knowledge, version can capture the whole field-of-view (FOV) of 50×30m m 2 at once and reduces the total imaging time down to 30min, while preserving the same high resolution (∼1.3m m) and superior quantitative accuracy (<7% error) as the previous versions. Therefore, this new method is an important step toward utilization of TMFT for preclinical imaging.

Optical Breast Imaging: A Review of Physical Principles, Technologies, and Clinical Applications.

Optical imaging involves the propagation of light through tissue. Current optical breast imaging technologies, including diffuse optical spectroscopy, diffuse optical tomography, and photoacoustic imaging, capitalize on the selective absorption of light in the near-infrared spectrum by deoxygenated and oxygenated hemoglobin. They provide information on the morphological and functional characteristics of different tissues based on their varied interactions with light, including physiologic information on lesion vascular content and anatomic information on tissue vascularity. Fluorescent contrast agents, such as indocyanine green, are used to visualize specific tissues, molecules, or proteins depending on how and where the agent accumulates. In this review, we describe the physical principles, spectrum of technologies, and clinical applications of the most common optical systems currently being used or developed for breast imaging. Most notably, US co-registered photoacoustic imaging and US-guided diffuse optical tomography have demonstrated efficacy in differentiating benign from malignant breast masses, thereby improving the specificity of diagnostic imaging. Diffuse optical tomography and diffuse optical spectroscopy have shown promise in assessing treatment response to preoperative systemic therapy, and photoacoustic imaging and diffuse optical tomography may help predict tumor phenotype. Lastly, fluorescent imaging using indocyanine green dye performs comparably to radioisotope mapping of sentinel lymph nodes and appears to improve the outcomes of autologous tissue flap breast reconstruction.

Preparation of polyacrylonitrile-based solution blow spinning fluorescent nanofibers from perylene-doped silica nanoparticles

ABSTRACT Fluorescent agents have been used to improve the anticounterfeiting of marketable products. It has been vital to enhance the engineering processing of fluorescent agents and materials matrices to prepare a mechanically reliable nanocomposite material with anticounterfeiting properties. Herein, new fluorescent and mechanically reliable inorganic-organic hybrid nanofibrous were developed by solution blowing spinning of polyacrylonitrile (PAN) encapsulating perylene-doped silica nanoparticles (PSiN). Two different types of fluorescent nanofibers were prepared using PSiN as an active doping agent and polyacrylonitrile as a hosting material. Fluorescent nanofibers were provided by solution blowing spinning technology utilizing perylene pigments, including perylene potassium (PMIK) and mono-imide sodium (PMINa) salts, and perylene diimide (PDI). In order to create a transparent nanofibrous film, fluorescent perylene-doped silica nanoparticles must be efficiently dispersed in a polyacrylonitrile solution without aggregation. The prepared fluorescent silica nanoparticles (SiN) displayed diameters ranging between 45 and 65 nm. The provided solution blow spun fluorescent PAN nanofibers integrated with PMINa, PMIK or PDI, showed smooth surface with diameters ranging between 350 and 550 nm. The prepared fluorescent PSiN and PSiN@PAN nanofibers were examined by several techniques, including transmission electron microscopy (TEM), scanning electron microscopy (SEM), and infrared spectroscopy (FT-IR). The solution blow spun nanofibers demonstrated an absorbance band at 458 nm and a fluorescence peak at 647 nm. The prepared nanofibers displayed a reversible emission upon repeated illumination with UV light. The emission properties of the prepared PSiN@PAN nanofibers were studied. The cytotoxic properties of the nanofibrous films were also explored. The current strategy introduces fluorescent nanostructured fibers that pave the way for the preparation of optical textiles such as optical devices, anticounterfeiting applications, and flexible displays.

Imaging and monitoring of granzyme B in the immune response.

Significant progress has been made in tumor immunotherapy that uses the human immune response to kill and remove tumor cells. However, overreactive immune response could lead to various autoimmune diseases and acute rejection. Accurate and specific monitoring of immune responses in these processes could help select appropriate therapies and regimens for the patient and could reduce the risk of side effects. Granzyme B (GzmB) is an ideal biomarker for immune response, and its peptide substrate could be coupled with fluorescent dyes or contrast agents for the synthesis of imaging probes activated by GzmB. These small molecules and nanoprobes based on PET, bioluminescence imaging, or fluorescence imaging have proved to be highly GzmB specific and accuracy. This review summarizes the design of different GzmB-responsive imaging probes and their applications in monitoring of tumor immunotherapy and overreactive immune response. This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging.

Safety assessment of fluorescently labeled anti-EGFR Nanobodies in healthy dogs.

Introduction: Surgical resection is one of the main treatment options for several types of cancer, the desired outcome being complete removal of the primary tumor and its local metastases. Any malignant tissue that remains after surgery may lead to relapsing disease, negatively impacting the patient's quality of life and overall survival. Fluorescence imaging in surgical oncology aims to facilitate full resection of solid tumors through the visualization of malignant tissue during surgery, following the administration of a fluorescent contrast agent. An important class of targeting molecules are Nanobodies® (Nbs), small antigen-binding fragments derived from camelid heavy chain only antibodies. When coupled with a fluorophore, Nbs can bind to a specific receptor and demarcate tumor margins through a fluorescence camera, improving the accuracy of surgical intervention. A widely investigated target for fluorescence-guided surgery is the epidermal growth factor receptor (EGFR), which is overexpressed in several types of tumors. Promising results with the fluorescently labeled anti-EGFR Nb 7D12-s775z in murine models motivated a project employing the compound in a pioneering study in dogs with spontaneous cancer. Methods: To determine the safety profile of the study drug, three healthy purpose-bred dogs received an intravenous injection of the tracer at 5.83, 11.66, and 19.47 mg/m2, separated by a 14-day wash-out period. Physical examination and fluorescence imaging were performed at established time points, and the animals were closely monitored between doses. Blood and urine values were analyzed pre- and 24 h post administration. Results: No adverse effects were observed, and blood and urine values stayed within the reference range. Images of the oral mucosa, acquired with a fluorescence imaging device (Fluobeam®), suggest rapid clearance, which was in accordance with previous in vivo studies. Discussion: These are the first results to indicate that 7D12-s775z is well tolerated in dogs and paves the way to conduct clinical trials in canine patients with EGFR-overexpressing spontaneous tumors.

Shades of phototoxicity in fluorescent imaging agents (that are not supposed to be phototoxic).

This article is a highlight of the paper by Huang et al. in this issue of Photochemistry and Photobiology. It describes shades of phototoxicity in fluorescent imaging agents that are not intended to be phototoxic. Phototoxicity was assessed using a modified neutral red uptake (NRU) in vitro assay with mean photo-effects (MPE) for the fluorescent agents IRdye800, indocyanine green (ICG), proflavine, and methylene blue (MB), with comparisons to known phototoxic agents benzoporphyrin derivative (BPD) and rose bengal (RB). The experimental conditions were aimed to mimic clinical settings, using not only visible light, but also near-infrared light for insight to photosafety and deep tissue damage. Molecular mechanisms underlying the phototoxicities were not sought, but IRdye800 and ICG were mainly deemed to be safe, whereas proflavine and MB would require precautions since phototoxicity can overshadow their utility as fluorescent imaging agents.