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Optical Imaging of Triple-Negative Breast Cancer Cells in Xenograft Athymic Mice Using an ICAM-1-Targeting Small-Molecule Probe
摘要： Purpose: The development of early, accurate diagnostic strategies for triple-negative breast cancer (TNBC) remains a significant challenge. Intercellular adhesion molecule-1 (ICAM-1) overexpressed in human TNBC cells is a potential molecular target and biomarker for diagnosis. In this study, small-molecule probe (denoted as γ3-Cy5.5) constructed with a short 17-mer linear peptide (γ3) and near-infrared fluorescence (NIRF) dye cyanine 5.5 (Cy5.5) was used to detect the expression of ICAM-1 in vitro and in vivo, and to diagnose TNBC via NIRF imaging. Procedures: Western blotting and flow cytometric analysis were used for the detection of ICAM-1 expression in MDA-MB-231 and MCF-7 cells. The cytotoxicity of the small-molecule probe γ3-Cy5.5 was detected using the CCK8 assay. The in vitro targeting of the small-molecule probe γ3-Cy5.5 was verified via flow cytometry and a laser scanning confocal microscope. Finally, the targeting of small-molecule probe in vivo and ex vivo was observed by NIRF imaging. Results: Western blotting and flow cytometry demonstrate that ICAM-1 was highly expressed in the MDA-MB-231 TNBC cell line. Laser confocal microscopy and flow cytometry results show that TNBC cells have an increased cellular uptake of γ3-Cy5.5 compared to the control probe γ3S-Cy5.5. With in vivo NIRF, a significantly higher Cy5.5 signal appeared in the tumors of mice administered γ3-Cy5.5 than those treated with γ3S-Cy5.5. The target-to-background ratio observed on the NIRF images was significantly higher in the γ3-Cy5.5 group (10.2, 13.6) compared with the γ3S-Cy5.5 group (4.4, 4.0) at 1 and 2 h, respectively. Conclusions: This is the first report of the use of ICAM-1-specific small-molecule probe for in vivo NIRF optical imaging of TNBC. This method provides a noninvasive and specific strategy for the early diagnosis of TNBC.
关键词： Triple-negative breast cancer,Near-infrared fluorescence imaging,Small-molecule probe,ICAM-1
更新于2025-09-23 15:23:52
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Sensitive detection and imaging of endogenous peroxynitrite using a benzo[d]thiazole derived cyanine probe
摘要： Peroxynitrite is a short-lived endogenous reactive species and plays important roles in many physiological and pathological processes. In this work, we synthesized a near-infrared probe based on the structure of benzothiazole derived cyanine for determination of peroxynitrite (ONOO-). The designed probe specifically reacted with ONOO- through oxidative cleavage of conjugated C?C double bonds and generating the non-fluorescent product. Meanwhile, the characteristic absorption of the probe at 630 nm greatly decreased after reaction with ONOO-, accompanied by drastic color change from bright blue to green yellow, which exhibited a distinct visual feature. It was demonstrated that the probe could be used to measure ONOO- in a dose-response manner and had a detection limit lower as 26 nM. Furthermore, the probe Cy-SN was applied for the imaging of endogenous ONOO- in living cells by confocal microscopy, which showed good cell permeability and low cytotoxicity. Successful application of probe for exogenous colorimetric detection and endogenous fluorescence imaging of ONOO- is suggesting its great potential applications in biological analysis.
关键词： Colorimetric,Peroxynitrite,Benzothiazole derived cyanine,Fluorescent probe,Endogenous fluorescence imaging
更新于2025-09-23 15:23:52
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Research on indocyanine green angiography for predicting postoperative hypoparathyroidism
摘要： Background: It may be critical to locate the parathyroid for surgeons during thyroidectomy or parathyroidectomy due to the significant function of the parathyroid in calcium balance. According to recent reports, the intrinsic fluorescence of the parathyroid has been found. There is some evidence supporting that new equipment can detect fluorescence via imaging technology. In this study, a newly-invented intraoperative fluorescence imaging system and indocyanine green dye were applied to detect the parathyroid glands and evaluate the vascularization of the parathyroid. The report is as follows. Methods: From May 1st to August 8st, 2018, 26 patients underwent total thyroidectomy in Zhuhai People’s Hospital and were recruited into our research. All identified parathyroid glands were scored visually from grade 0 to grade 2 according to the vascularity of the parathyroid before ICG angiography was performed. After ICG angiography, parathyroid glands were scored from score 0 to score 2 according to the FI. Results: Visual scores were significantly higher than ICG angiography scores. In the 22 patients with at least one parathyroid gland with an ICG score of 2, postoperative PTH levels were in the normal range. In the 4 patients with no parathyroid gland with an ICG score of 2, 2 of them developed transient hypoparathyroidism, with recovery on POD 7 for the first patient and after 3 months for the second one. Conclusion: This study has identified that the fluorescence imaging system applied with indocyanine green is a safe, easy and effective method to protect the parathyroid and predict postoperative hypoparathyroidism. Registration number: ChiCTR1800016864.
关键词： Fluorescence imaging system,Indocyanine green,Postoperative hypoparathyroidism.,Parathyroid
更新于2025-09-23 15:23:52
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A Fluorescent Cocktail Strategy for Differentiating Tumor, Inflammation, and Normal Cells by Detecting mRNA and H <sub/>2</sub> O <sub/>2</sub>
摘要： Accurately distinguishing tumors from noncancerous inflammation and normal tissues is hugely significant for tumor diagnosis and therapy. However, tumor and inflammatory tissues have similar pathologic characteristics in their microenvironment, making differentiation very difficult. Here, a fluorescent cocktail nanoparticle capable of simultaneously detecting intracellular mRNA and H2O2 was designed to differentiate tumors from nontumor cells. To detect targeted mRNA in living cells, a DNA probe was generated using the fluorescence resonance energy transfer (FRET) principle. A pH-responsive amphiphilic polymer was synthesized to realize the transportation of the DNA probe. In addition, the polymer was conjugated with a coumarin-boronic acid ester (Cou-BE) H2O2 probe. According to the change in the fluorescence of Cou-BE, tumor and inflammatory cells could be distinguished from normal cells owing to their high concentration of H2O2. Because of the different concentrations of tumor-related mRNA in tumor and nontumor cells, the fluorescence intensity of the DNA probe-loaded nanoparticles inside tumor cells was different from that inside inflammatory cells. Therefore, our fluorescent cocktail strategy could discriminate simultaneously tumor, inflammation, and normal cells through the cooperative detection of intracellular mRNA and H2O2, which demonstrated potential application value in biomedical research and clinical diagnosis.
关键词： H2O2,tumor,tumor-related mRNA,fluorescence imaging,inflammation
更新于2025-09-23 15:23:52
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Observation of Acetylcholinesterase in Stress-induced Depression Phenotypes by Two-photon Fluorescence Imaging in the Mouse Brain
摘要： Oxidative stress in depression is a prime cause of neurotransmitter metabolism dysfunction in the brain. Acetylcholinesterase (AChE), a key hydrolase in the cholinergic system, directly determines the degradation of neurotransmitters. However, due to the complexity of the brain and lack of appropriate in situ imaging tools, the mechanism underlying the changes in AChE activity in depression remains unclear. Hence, we generated a two-photon fluorescence probe (MCYN) for real-time visualization of AChE with excellent sensitivity and selectivity. AChE can specifically recognize and cleave the carbamic acid ester bond in MCYN, and MCYN emits bright fluorescence at 560 nm by two-photon excitation at 800 nm. By utilizing MCYN to monitor AChE, we discovered a significant increase in AChE activity in the brains of mice with depression phenotypes. Notably, with the assistance of a two-photon fluorescence imaging probe of the superoxide anion radical (O2??), in vivo visualization for the first time revealed the positive correlation between AChE and O2?? levels associated with depressive behaviors. This finding suggests that oxidative stress may induce AChE overactivation, leading to depression-related behaviors. This work provides a new and rewarding perspective to elucidate the role of oxidative stress regulating AChE in the pathology of depression.
关键词： acetylcholinesterase,depression,Two-photon fluorescence imaging,brain,oxidative stress
更新于2025-09-23 15:23:52
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Yellow-emissive carbon dots with a large Stokes shift are viable fluorescent probes for detection and cellular imaging of?silver ions and glutathione
摘要： Yellow-emissive carbon dots (Y-CDs) were prepared by a solvothermal method using anhydrous citric acid and 2,3-phenazinediamine as the starting materials. The Y-CDs display a 24% fluorescence quantum yield, a 188-nm Stokes’ shift and excellent stability. They are shown here to be excellent fluorescent probes for the determination of Ag(I) ion and glutathione (GSH). If exposed to Ag(I) ions, they are bound by the carboxy groups of the Y-CDs, and this causes quenching of fluorescence (with excitation/emission maxima at 380/568 nm) via a static quenching mechanism. This effect was used to design a fluorometric assay for Ag(I). The quenched fluorescence of the Y-CDs can be restored by adding GSH due to the high affinity of GSH for Ag(I). The calibration plot for Ag(I) is linear in the 1–4 μM Ag(I) concentration range, and the limit of detection is 31 nM. The respective values for GSH are 5–32 μM, and 76 nM, respectively. The method was applied to the detection of Ag(I) in spiked environmental water samples and gave recoveries ranging from 93 to 107%. It was also applied to the determination of GSH in tomatoes and purple grapes and gave satisfactory recoveries. The Y-CDs display low cytotoxicity and were successfully used to image Ag(I) and GSH in H1299 cells.
关键词： Fluorescence microscopy,Applications,Stern-Volmer plot,Fluorescence imaging,Fluorescence detection
更新于2025-09-23 15:22:29
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Doxorubicin and indocyanine green loaded superparamagnetic iron oxide nanoparticles with PEGylated phospholipid coating for magnetic resonance with fluorescence imaging and chemotherapy of glioma
摘要： Background: Glioma represents the most common malignant brain tumor. Outcomes of surgical resection are often unsatisfactory due to low sensitivity or resolution of imaging methods. Moreover, the use of traditional chemotherapeutics, such as doxorubicin (DOX), is limited due to their low blood–brain barrier (BBB) permeability. Recently, the development of nanotechnology could overcome these obstacles. Materials and methods: Hydrophobic superparamagnetic iron oxide nanoparticles (SPIO NPs) were prepared with the use of thermal decomposition method. They were coated with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG 2000) and DOX using a thin-film hydration method followed by loading of indocyanine green (ICG) into the phospholipid layers. Details regarding the characteristics of NPs were determined. The in vitro biocompatibility and antitumor efficacy were established with the use of MTT assay. In vivo fluorescence and magnetic resonance (MR) imaging were used to evaluate BBB penetration and accumulation of NPs at the tumor site. Antitumor efficacy was evaluated using measures of tumor size, median survival times, body weights, and H&E staining. Results: The multifunctional NPs generated had an average diameter of 22.9 nm, a zeta potential of -38.19 mV, and were capable of providing a sustained release of DOX. In vitro experiments demonstrated that the SPIO@DSPE-PEG/DOX/ICG NPs effectively enhanced cellular uptake of DOX as compared with that of free DOX. In vivo fluorescence and MR imaging revealed that the NPs not only effectively crossed the BBB but selectively accumulated at the tumor site. Meanwhile, among all groups studied, C6 glioma-bearing rats treated with the NPs exhibited the maximal degree of therapeutic efficacy, including smallest tumor volume, lowest body weight loss, and longest survival times, with no obvious side effects. Conclusion: These results suggest that the SPIO@DSPE-PEG/DOX/ICG NPs can not only function as a nanoprobe for MR and fluorescence bimodal imaging, but also as a vehicle to deliver chemotherapeutic drugs to the tumor site, to achieve the theranostic treatment of glioma.
关键词： SPIO NPs,fluorescence imaging,chemotherapy,BBB,MR imaging
更新于2025-09-23 15:22:29
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[IEEE 2018 26th European Signal Processing Conference (EUSIPCO) - Rome (2018.9.3-2018.9.7)] 2018 26th European Signal Processing Conference (EUSIPCO) - DeepMQ: A Deep Learning Approach Based Myelin Quantification in Microscopic Fluorescence Images
摘要： Oligodendrocytes wrap around the axons and form the myelin. Myelin facilitates rapid neural signal transmission. Any damage to myelin disrupts neuronal communication leading to neurological diseases such as multiple sclerosis (MS). There is no cure for MS. This is, in part, due to lack of an efficient method for myelin quantification during drug screening. In this study, an image analysis based myelin sheath detection method, DeepMQ, is developed. The method consists of a feature extraction step followed by a deep learning based binary classification module. The images, which were acquired on a confocal microscope contain three channels and multiple z-sections. Each channel represents either oligodendroyctes, neurons, or nuclei. During feature extraction, 26-neighbours of each voxel is mapped onto a 2D feature image. This image is, then, fed to the deep learning classifier, in order to detect myelin. Results indicate that 93.38% accuracy is achieved in a set of fluorescence microscope images of mouse stem cell-derived oligodendroyctes and neurons. To the best of authors’ knowledge, this is the first study utilizing image analysis along with machine learning techniques to quantify myelination.
关键词： neural network,microscopic fluorescence imaging,myelin,deep learning,LeNet
更新于2025-09-23 15:22:29
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Dual-Modal In Vivo Fluorescence/Photoacoustic Microscopy Imaging of Inflammation Induced by GFP-Expressing Bacteria
摘要： In this study, dual-modal fluorescence and photoacoustic microscopy was performed for noninvasive and functional in vivo imaging of inflammation induced by green fluorescent protein (GFP) transfected bacteria in mice ear. Our imaging results demonstrated that the multimodal imaging technique is able to monitor the tissue immunovascular responses to infections with molecular specificity. Our study also indicated that the combination of photoacoustic and fluorescence microscopy imaging can simultaneously track the biochemical changes including the bacterial distribution and morphological change of blood vessels in the biological tissues with high resolution and enhanced sensitivity. Consequently, the developed method paves a new avenue for improving the understanding of the pathology mechanism of inflammation.
关键词： GFP-expressing bacteria,fluorescence imaging,biosensor,photoacoustic microscopy,inflammation/infection
更新于2025-09-23 15:22:29
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Indocyanine green-based fluorescence imaging in visceral and hepatobiliary and pancreatic surgery: State of the art and future directions
摘要： In recent years, the use of fluorescence-guided surgery (FGS) to treat benign and malignant visceral, hepatobiliary and pancreatic neoplasms has significantly increased. FGS relies on the fluorescence signal emitted by injected substances (fluorophores) after being illuminated by ad hoc laser sources to help guide the surgical procedure and provide the surgeon with real-time visualization of the fluorescent structures of interest that would be otherwise invisible. This review surveys and discusses the most common and emerging clinical applications of indocyanine green (ICG)-based fluorescence in visceral, hepatobiliary and pancreatic surgery. The analysis, findings, and discussion presented here rely on the authors’ significant experience with this technique in their medical institutions, an up-to-date review of the most relevant articles published on this topic between 2014 and 2018, and lengthy discussions with key opinion leaders in the field during recent conferences and congresses. For each application, the benefits and limitations of this technique, as well as applicable future directions, are described. The imaging of fluorescence emitted by ICG is a simple, fast, relatively inexpensive, and harmless tool with numerous different applications in surgery for both neoplasms and benign pathologies of the visceral and hepatobiliary systems. The ever-increasing availability of visual systems that can utilize this tool will transform some of these applications into the standard of care in the near future. Further studies are needed to evaluate the strengths and weaknesses of each application of ICG-based fluorescence imaging in abdominal surgery.
关键词： visceral perfusion,Peritoneal carcinomatosis,Pancreatic surgery,Indocyanine green,Liver surgery,Gastrointestinal surgery,Biliary surgery,Fluorescence imaging,Biliary anatomy
更新于2025-09-23 15:22:29