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Ratiometric fluorescence imaging for sodium selenite in living cells
摘要： Sodium selenite as a common selenium-containing agent has been extensively studied for human health, especially cancer prevention and treatment. However, precisely evaluating the dynamic changes of sodium selenite at cellular and molecular level in these pharmacological and pathophysiological studies is still challenging. Developing the chemical tools that enable the quantification of sodium selenite in vivo with high spatial and temporal fidelity thus can provide an efficient solution to this challenge. Here, we report the design, synthesis and biological evaluation of ratiometric sodium selenite fluorescent probe (HBTN-Se) for quantitative ratiometric fluorescence imaging of sodium selenite concentration fluctuations in living cells. A strategy for sodium selenite quantification has been achieved effectively through using a highly selective, rapid and biocompatible hydroxyl-deprotecting reaction. The ratiometric fluorescence imaging with HBTN-Se is constructed to precisely monitor changes in sodium selenite levels in living cells. Moreover, HBTN-Se is capable of visualizing differences in sodium selenite levels between normal cells and cancer cells, establishing the utility of this ratiometric detection platform for assessing the correlation between physiological effects of sodium selenite and its levels in living cells.
关键词： Quantitative determination,Fluorescent probe,Sodium selenite,Ratiometric fluorescence imaging
更新于2025-09-23 15:22:29
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A selective fluorescent chemosensor for Cd2+ based on 8-hydroxylquinoline-benzothiazole conjugate and imaging application
摘要： A 8-hydroxylquinoline-benzothiazole conjugate (HQ-BT) was facilely synthesized by two steps with more than 60% total reaction yield. The HQ-BT showed a weak fluorescence that could be strongly enhanced by coordination with various metal ions such as Al3+, Cd2+, Zn2+ in methanol containing 1% water. Interestingly, the selectivity toward Cd2+ was achieved by increasing water fraction to 30% aqueous methanol solution. Thus, the HQ-BT was developed as a new and selective fluorescent chemosensor for Cd2+ in aqueous solution with a broad pH region 4–12. A good linear relationship between the fluorescence intensity and the Cd2+ concentration was found in the range of 0–5 μM with a detection limit of 0.1 μM (S/N = 3). It was also succesfully used for fluorescence imaging of Cd2+ in living cells.
关键词： Benzothiazole,8-Hydroxylquinoline,Fluorescent chemosensor,Fluorescence imaging,Cd2+
更新于2025-09-23 15:22:29
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Precise anatomical sublobar resection using a 3D medical image analyzer and fluorescence-guided surgery with transbronchial instillation of indocyanine green
摘要： Objective: We developed a novel approach combined with 3D Image Analyzer and infrared thoracoscopy for pulmonary sublobar resection. The purpose of this study was to investigate the feasibility of this procedure. Methods: From October 2014 to April 2018, 65 cases were enrolled, and 58 cases were evaluated. For each case, several virtual sublobar resections were created by 3D Image Analyzer preoperatively. The surgical margin was measured in each simulated sublobar resection and the most appropriate procedure was selected. Surgical resection with matching virtual sublobar resection was performed using infrared thoracoscopy with transbronchial indocyanine green instillation. We evaluated the border clarity of ICG fluorescence to investigate success of ICG injection and compared pre- and postoperative CTs to determine whether the correct area could be removed according to the simulation. We also compared short-term surgical outcomes between the ICG cases and historical segmentectomy cases by propensity score matching. Results: The success rate of transbronchial ICG injections was 89.2% (58/65). These 58 patients were eligible for evaluation of our procedure. Sublobar resection included subsegmental resection (5), simple segmentectomy (15), complex segmentectomy (16), and extended segmentectomy (22). The shortest distances to the surgical margin by simulation and by actual measurement were 21.5+/-11.2 mm and 23.5+/-8.3, respectively (p=0.190). Fifty-four of 58 cases underwent sublobar resection matched with the simulation (93.1% concordance rate). Operative results and short-term outcomes were similar between the two groups by propensity score matching. Conclusion: ICG-guided sublobar resection by transbronchial ICG instillation is feasible and applicable to any type of sublobar resection.
关键词： sublobar resection,fluorescence imaging,Synapse 3D,indocyanine green,lung neoplasms
更新于2025-09-23 15:22:29
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Rapid cancer diagnosis by highly fluorescent carbon nanodots-based imaging
摘要： Carbon dots (Cdots) with bright green fluorescence were applied to the rapid and selective cell imaging for a variety of cell lines. Different labeling distributions of hepatoma cells (HepG2) and normal human liver cells (LO2) were achieved using Cdots as imaging agents. For HepG2 cells, the Cdots could rapidly permeate the cell membrane and diffuse into the cytoplasm and nucleus within 3 min, and retained their location in the targets for 24 h. However, the Cdots exhibited bright fluorescence only in the cytoplasm of LO2 cell lines. Moreover, the Cdots were almost non-cytotoxic and exhibited superior photostability over a wide range of pH. Therefore, these Cdots have great potential for rapid, luminous and selective bioimaging applications, and are expected to be used as a nucleus-staining agent in cancer diagnosis.
关键词： Selective fluorescence imaging,Rapid nuclear targeting,Carbon dots
更新于2025-09-23 15:22:29
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The kinetic mechanisms of fast-decay red-fluorescent genetically-encoded calcium indicators
摘要： Genetically-encoded calcium indicators (GECIs) are useful reporters of cell signalling, neuronal and network activities. We have generated novel fast variants and investigated the kinetic mechanisms of recently developed red-fluorescent GECIs (RGECIs), mApple-based jRGECO1a and mRuby-based jRCaMP1a. In the formation of fluorescent jRGECO1a and jRCaMP1a complexes calcium binding is followed by rate-limiting isomerisation. However fluorescence decay of calcium-bound jRGECO1a follows a different pathway form its formation: dissociation of calcium occurs first followed by the peptide, similarly to GCaMP-s. In contrast, fluorescence decay of calcium-bound jRCaMP1a occurs by the on-pathway: peptide dissociation followed by calcium. The mechanistic differences explain the generally slower off-kinetics of jRCaMP1a-type indicators compared to GCaMP-s and jRGECO1a-type GECI: the fluorescence decay rate of f-RCaMP1 was 21 s-1, compared to 109 s-1 for f-RGECO1 and f-RGECO2 (37 °C). Thus, the CaM-peptide interface is an important determinant of the kinetic responses of GECIs, however the topology of the structural link to the fluorescent protein demonstrably affects internal dynamics of the CaM-peptide complex. In the dendrites of hippocampal CA3 neurons, f-RGECO1 indicates calcium elevation in response to a 100 action potential train in a linear fashion, making the probe particularly useful for monitoring large amplitude, fast signals e.g. those in dendrites, muscle and immune cells.
关键词： GECI,calcium,kinetics,fluorescence,imaging
更新于2025-09-23 15:22:29
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Near-infrared fluorescence laparoscopy of the ureter with three preclinical dyes in a pig model
摘要： Background Ureteric injury is reported to occur in 1–7.6% of colorectal surgeries. To reduce the incidence of ureteral injury, it is essential to identify the ureters. The use of near-infrared fluorescence (NIRF) imaging with intravenously administered dyes might be of added value for ureteral visualization during laparoscopy. The aim of this study is to assess the performance of three preclinical dyes; IRDye? 800BK, IRDye? 800NOS and IRDye? 800CW, for near-infrared fluorescence laparoscopy of the ureter in pigs. Methods In three female Dutch landrace pigs, the new dyes were evaluated. In each pig, 1 dye was tested using a 6-mg intravenous dose in a concentration of 1?mg/ml. Imaging was performed in fluorescence mode and white light mode with a laparoscopic imaging system. In order to further evaluate the dyes, an ex?vivo imaging experiment was performed, in which 8 decreasing concentrations per dye, diluted in PBS, were evaluated in a transparent test tube with NIRF mode at a distance of 1, 5 and 10?cm from the laparoscope. Results All three dyes were effective in allowing the identification of the ureter with NIRF imaging. The ureter became fluorescent after 35, 45 and 10?min, respectively, for IRDye? 800BK, IRDye? 800NOS and IRDye? 800CW with a maximum target-to-background ratio (TBR) of 2.14, 0.66 and 1.44, respectively. In the ex?vivo imaging experiment, all three dyes produced a strong fluorescence signal at all concentrations and all distances evaluated. Conclusions Intravenous administration of the preclinical dyes IRDye? 800CW, IRDye? 800 BK and IRDye? 800NOS facilitated successful identification of the anatomical course of the ureter in living pig models. The highest measured TBR occurred with the use of IRDye? 800BK. Ex?vivo, a correlation was observed between the fluorescence intensities of the signal with the concentration of the dye and with the distance to the object.
关键词： laparoscopic colorectal surgery,Near-infrared fluorescence imaging,Ureter,Fluorescent dyes
更新于2025-09-23 15:21:21
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Noninvasive In Situ Ratiometric Imaging of Biometals Based on Self-Assembled Peptide Nanoribbon
摘要： Development of probes for accurate sensing and imaging of biometals in situ is still a growing interest owing to their crucial roles in cellular metabolism, neurotransmission, and apoptosis. Among them, Zn2+ and Cu2+ are two important cooperative biometals closely related to Alzheimer’s disease (AD). Herein, we developed a multifunctional probe based on self-assembling peptide nanoribbon for ratiometric sensing of Zn2+, Cu2+, or Zn2+ and Cu2+ simultaneously. Uniform peptide nanoribbon (AQZ@NR) was rationally designed by coassembling a Zn2+-specific ligand AQZ-modified peptide (AQZKL-7) with peptide KL-7. The nanoribbon further combined with Cu2+-sensitive near-infrared quantum dots (NIR QDs) and Alexa Fluor 633 as an inner reference molecule, which was endowed with the capability for ratiometric Zn2+ and Cu2+ imaging at the same time. The peptide-based probe exhibited good specificity to Zn2+ and Cu2+ without interference from other ions. Importantly, the nanoprobe was successfully applied for noninvasive Zn2+ and Cu2+ monitoring in both living cells and zebrafish via multicolor fluorescence imaging. This gives insights into the dynamic Zn2+ and Cu2+ distribution in an intracellular and in vivo mode, as well as understanding the neurotoxicity of high concentration of Zn2+ and Cu2+. Therefore, the self-assembled nanoprobe shows great promise in multiplexed detection of many other biometals and biomolecules, which will benefit the diagnosis and treatment of AD in clinical applications.
关键词： fluorescence imaging,biometals,ratiometric imaging,peptide nanoribbon,Cu2+,Zn2+,self-assembly,Alzheimer’s disease
更新于2025-09-23 15:21:01
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Aprotinin Encapsulated Gold Nanoclusters: A Fluorescent Bioprobe with Dynamically Nuclear Targeting and Selective Detection of Trypsin and Heavy Metal
摘要： Fluorescence imaging has currently emerged as one of the most frequently used noninvasive imaging technology to selectively monitor biological processes in living systems. In past decades, gold nanoclusters (Au NCs) received increasing attraction because of their intrinsic fluorescence and their inherent biocompatibility. As a stabilizing and reducing agent, an abundant, sustainable and widely used polypeptide derived drug molecule, aprotinin (Ap) is selected for the synthesis of Au nanoclusters (Ap-Au NCs) due to characteristic bioactivity, excellent biocompatibility, biodegradability, nonallergenic character. Herein, Ap encapsulated Au NCs exhibiting desirable red fluorescence feature was facilely produced for the first time, which were subsequently used for cell imaging and detection of various analytes. Much interestingly, dynamically subcellular localization of Ap-Au NCs from the cytoplasm to the nucleus in Hela cells was observed. Afterward, it has shown the selective and quantitative detection of trypsin by using Ap encapsulated Au NCs. Lastly, Ap-Au NCs were readily used for detection of mercury and copper quantitatively. The photoluminescence of the Ap-Au NCs was quenched with the addition of metioned analytes. This study opens crucial insights on the integration of biomolecule with metal nanoclusters, also discusses a multifunctional nanomaterial platform for cell imaging, subcellular targeting, biosensing and drug delivery.
关键词： trypsin detection,gold nanoclusters,aprotinin,cell imaging,heavy metal detection,Fluorescence imaging
更新于2025-09-23 15:21:01
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Staging laparoscopy with ultrasound and near-infrared fluorescence imaging to detect occult metastases of pancreatic and periampullary cancer
摘要： Introduction Up to 38% of pancreatic and periampullary cancer patients undergoing curative intended surgery turn out to have incurable disease. Therefore, staging laparoscopy (SL) prior to laparotomy is advised to spare patients the morbidity, inconvenience and expense of futile major surgery. The aim of this study was to assess the added value of SL with laparoscopic ultrasonography (LUS) and laparoscopic near-infrared fluorescence imaging (LFI). Methods All patients undergoing curative intended surgery of pancreatic or periampullary cancer were included prospectively in this single arm study. Patients received an intravenous infusion of 10 mg indocyanine green (ICG) one or two days prior to surgery to allow LFI. Suspect lesions were analyzed via biopsy or resection. Follow-up visits after surgery occurred every three months. Results A total of 25 patients were included. Suspect lesions were identified in 7 patients: liver metastases (n = 2; identified by inspection, LUS, and LFI), peritoneal metastases (n = 1; identified by inspection only), and benign lesions (n = 4; identified by inspection or LUS). Quality of LFI was good in 67% (10/15) of patients dosed one day and 89% (8/9) dosed two days prior to surgery. A futile laparotomy was averted in 3 patients (12%). Following SL the primary tumor was resected in 20 patients. Two patients (10%) developed metastases within 3 months after resection. Conclusions Despite current preoperative imaging modalities metastases are still identified during surgery. This study shows limited added value of LUS during SL in patients with pancreatic or periampullary cancer. LFI was of added value due to its high negative predictive value in case of suspect hepatic lesions identified by inspection.
关键词： near-infrared fluorescence imaging,staging laparoscopy,laparoscopic ultrasonography,pancreatic cancer,periampullary cancer,indocyanine green
更新于2025-09-23 15:21:01
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NIR Infrared imaging after peritumoral injection of indocyanine green to guide lymph node dissection in head and neck squamous cell carcinoma: A pilot feasibility study
摘要： In head and neck squamous cell carcinoma (HNSCC), neck dissection is part of the surgical therapy. Beyond to the fact that radicality is a critical prognostic factor, this dissection could lead to significant morbidity, potentially avoidable when resected lymph nodes (LN) are proven to be non-invaded at pathology. Therefore, the definition of a method able to reliably identify the specific LN drainage area in HNSCC would represent a relevant progress, to better guide the neck dissection, potentially improving the radicality and reducing the morbidity. As near-infrared fluorescence imaging (NIR-FI) after indocyanine green (ICG) peritumoral injection has been validated as a sentinel procedure, we hypothesized that this approach could represent a new technique to identify the tumor-drainage area in HNSCC. We prospectively evaluated this technique in 14 patients with oral or oropharyngeal carcinoma scheduled for primary tumor resection and LN dissection. The trial was approved by the ethics committee of the Institut Jules Bordet (CE-2178) (EudraCT 2014-000298-37) and all patients signed informed consent before inclusion.
关键词： Head and neck squamous cell carcinoma,Indocyanine green,Near-infrared fluorescence imaging,Lymph node dissection,Surgical guidance
更新于2025-09-23 15:21:01