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Search Results (736)

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Keywords = fluorescence imaging probes

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12 pages, 1076 KiB  
Article
Rapid Identification of the SNP Mutation in the ABCD4 Gene and Its Association with Multi-Vertebrae Phenotypes in Ujimqin Sheep Using TaqMan-MGB Technology
by Yue Zhang, Min Zhang, Hong Su, Jun Liu, Feifei Zhao, Yifan Zhao, Xiunan Li, Yanyan Yang, Guifang Cao and Yong Zhang
Animals 2025, 15(15), 2284; https://doi.org/10.3390/ani15152284 - 5 Aug 2025
Abstract
Ujimqin sheep, known for its distinctive multi-vertebrae phenotypes (T13L7, T14L6, and T14L7) and economic value, has garnered significant attention. However, conventional phenotypic detection methods suffer from low efficiency and high costs. In this study, based on a key SNP locus (ABCD4 gene, [...] Read more.
Ujimqin sheep, known for its distinctive multi-vertebrae phenotypes (T13L7, T14L6, and T14L7) and economic value, has garnered significant attention. However, conventional phenotypic detection methods suffer from low efficiency and high costs. In this study, based on a key SNP locus (ABCD4 gene, Chr7:89393414, C > T) identified through a genome-wide association study (GWAS), a TaqMan-MGB (minor groove binder) genotyping system was developed. the objective was to establish a high-throughput and efficient molecular marker-assisted selection (MAS) tool. Specific primers and dual fluorescent probes were designed to optimize the reaction system. Standard plasmids were adopted to validate genotyping accuracy. A total of 152 Ujimqin sheep were subjected to TaqMan-MGB genotyping, digital radiography (DR) imaging, and Sanger sequencing. the results showed complete concordance between TaqMan-MGB and Sanger sequencing, with an overall agreement rate of 83.6% with DR imaging. For individuals with T/T genotypes (127/139), the detection accuracy reached 91.4%. This method demonstrated high specificity, simplicity, and cost-efficiency, significantly reducing the time and financial burden associated with traditional imaging-based approaches. the findings indicate that the TaqMan-MGB technique can accurately identify the T/T genotype at the SNP site and its strong association with the multi-vertebrae phenotypes, offering an effective and reliable tool for molecular breeding of Ujimqin sheep. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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13 pages, 1484 KiB  
Article
A Long-Wavelength Fluorescent Probe for Efficient Dual-Color Imaging of Boronic-Acid-Containing Agents in Living Cells
by Shinya Takada, Honghuo Du, Naoya Kondo, Anna Miyazaki, Fumiko Hara, Shizuyo Horiyama, Takashi Temma and Masayori Hagimori
Chemosensors 2025, 13(8), 283; https://doi.org/10.3390/chemosensors13080283 - 4 Aug 2025
Viewed by 126
Abstract
In boron neutron capture therapy (BNCT), the intracellular localization and concentration of boron-10 atoms significantly influence therapeutic efficacy. Although various boronic-acid-targeted fluorescent probes have been developed to evaluate BNCT agents, most of these probes emit at short wavelengths and are, therefore, incompatible with [...] Read more.
In boron neutron capture therapy (BNCT), the intracellular localization and concentration of boron-10 atoms significantly influence therapeutic efficacy. Although various boronic-acid-targeted fluorescent probes have been developed to evaluate BNCT agents, most of these probes emit at short wavelengths and are, therefore, incompatible with common nuclear-staining reagents such as Hoechst 33342 and 4′,6-diamidino-2-phenylindole (DAPI). While our previously reported probe, BS-631, emitted fluorescence above 500 nm, it exhibited limitations in terms of reaction rate and fluorescence intensity. To address these issues, we developed a boronic-acid-targeted fluorescent probe with a longer emission wavelength, rapid reactivity, and strong fluorescence intensity. Herein, we designed and synthesized BTTQ, a probe based on a 2-(2-hydroxyphenyl)benzothiazole core structure. BTTQ exhibited immediate fluorescence upon reaction with 4-borono-L-phenylalanine (BPA), with an emission wavelength of 567 nm and a sufficiently high fluorescence quantum yield for detection. BTTQ quantitatively detected BPA with high sensitivity (quantification limit of 10.27 µM), suitable for evaluating BNCT agents. In addition, BTTQ exhibited selective fluorescence for BPA over metal cations. Importantly, BTTQ enabled fluorescence microscopic imaging of intracellular BPA distribution in living cells co-stained with Hoechst 33342. These results suggest that BTTQ is a promising fluorescent probe for the evaluation of future BNCT agents. Full article
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21 pages, 2240 KiB  
Review
A Review of Fluorescent pH Probes: Ratiometric Strategies, Extreme pH Sensing, and Multifunctional Utility
by Weiqiao Xu, Zhenting Ma, Qixin Tian, Yuanqing Chen, Qiumei Jiang and Liang Fan
Chemosensors 2025, 13(8), 280; https://doi.org/10.3390/chemosensors13080280 - 2 Aug 2025
Viewed by 205
Abstract
pH is a critical parameter requiring precise monitoring across scientific, industrial, and biological domains. Fluorescent pH probes offer a powerful alternative to traditional methods (e.g., electrodes, indicators), overcoming limitations in miniaturization, long-term stability, and electromagnetic interference. By utilizing photophysical mechanisms—including intramolecular charge transfer [...] Read more.
pH is a critical parameter requiring precise monitoring across scientific, industrial, and biological domains. Fluorescent pH probes offer a powerful alternative to traditional methods (e.g., electrodes, indicators), overcoming limitations in miniaturization, long-term stability, and electromagnetic interference. By utilizing photophysical mechanisms—including intramolecular charge transfer (ICT), photoinduced electron transfer (PET), and fluorescence resonance energy transfer (FRET)—these probes enable high-sensitivity, reusable, and biocompatible sensing. This review systematically details recent advances, categorizing probes by operational pH range: strongly acidic (0–3), weakly acidic (3–7), strongly alkaline (>12), weakly alkaline (7–11), near-neutral (6–8), and wide-dynamic range. Innovations such as ratiometric detection, organelle-specific targeting (lysosomes, mitochondria), smartphone colorimetry, and dual-analyte response (e.g., pH + Al3+/CN) are highlighted. Applications span real-time cellular imaging (HeLa cells, zebrafish, mice), food quality assessment, environmental monitoring, and industrial diagnostics (e.g., concrete pH). Persistent challenges include extreme-pH sensing (notably alkalinity), photobleaching, dye leakage, and environmental resilience. Future research should prioritize broadening functional pH ranges, enhancing probe stability, and developing wide-range sensing strategies to advance deployment in commercial and industrial online monitoring platforms. Full article
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16 pages, 6426 KiB  
Article
Manganese-Rich Chromite in Myanmar Jadeite Jade: A Critical Source of Chromium and Manganese and Its Role in Coloration
by Yu Zhang, Guanghai Shi and Jiabao Wen
Crystals 2025, 15(8), 704; https://doi.org/10.3390/cryst15080704 - 31 Jul 2025
Viewed by 177
Abstract
Color is a primary determinant of the value of jadeite jade, but the petrological provenance of the chromogenic elements of jadeite jade remains uncertain. The characteristics of the associated chromite in Myanmar jadeite jade were systematically investigated through a series of tests, including [...] Read more.
Color is a primary determinant of the value of jadeite jade, but the petrological provenance of the chromogenic elements of jadeite jade remains uncertain. The characteristics of the associated chromite in Myanmar jadeite jade were systematically investigated through a series of tests, including polarized microscopy, microarea X-ray fluorescence spectroscopy (micro-XRF) mapping, electron probe microanalysis (EPMA), and backscattered electron (BSE) imaging. The results demonstrate that the chromite composition in Myanmar jadeite jade is characterized by a high concentration of Cr2O3 (46.18–67.11 wt.%), along with a notable abundance of MnO (1.68–9.13 wt.%) compared with the chromite from the adjacent Myitkyina peridotite. The diffusion of chromium (Cr) and manganese (Mn) in jadeite jade is accomplished by accompanying the metamorphic pathway of Mn-rich chromite → kosmochlor → chromian jadeite → jadeite. In the subsequent phase of jadeite jade formation, the chromium-rich omphacite veins generated by the fluid enriched in Ca and Mg along the fissures of kosmochlor and chromian jadeite play a role in the physical diffusion of Cr and Mn. The emergence of the lavender hue in jadeite is contingent upon the presence of a relatively high concentration of Mn (approximately 100–1000 ppmw) and the simultaneous absence of Cr, which would otherwise serve as a more effective chromophore (no Cr or up to a dozen ppmw). The distinctive Mn-rich chromite represents the primary origin of the chromogenic element Cr (green) and, perhaps more notably, an overlooked provider of Mn (lavender) in Myanmar jadeite jade. Full article
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18 pages, 3972 KiB  
Article
Naphthalimide-Based Fluorescent Probe for Portable and Rapid Response to γ-Glutamyl Transpeptidase
by Jinhu Wang, Xianchao Jia, Yihao Zhang, Ye Gao, Lei Zhang, Changgong Meng, Zhaohui Wang and Yang Jiao
Molecules 2025, 30(15), 3174; https://doi.org/10.3390/molecules30153174 - 29 Jul 2025
Viewed by 226
Abstract
γ-Glutamyl transpeptidase (GGT) is overexpressed in a variety of diseases, making it an important diagnostic criterion for diseases. Herein, a new fluorescence probe based on naphthalimide (Glu-MDA) was developed and employed for the rapid detection of GGT in tumor cells or samples. Alkynylated [...] Read more.
γ-Glutamyl transpeptidase (GGT) is overexpressed in a variety of diseases, making it an important diagnostic criterion for diseases. Herein, a new fluorescence probe based on naphthalimide (Glu-MDA) was developed and employed for the rapid detection of GGT in tumor cells or samples. Alkynylated naphthalimide is the fluorescent core for excellent fluorescence response. The covalent bridging of self-immolative short linkers reduces the steric hindrance between probes and enzyme cleavage sites, which leads to improved enzymatic reaction kinetics. Glu-MDA shows a rapid response and excellent selectivity with a detection limit of 0.044 U/L. This allows the efficient detection of GGT levels in solution and cells. Simultaneously, the construction of Glu-MDA pre-stained test strips provided an innovative strategy for the qualitative detection of GGT activity, helping to detect GGT faster, more portably, and cost-effectively in various scenarios. Full article
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17 pages, 4161 KiB  
Article
Targeting CEACAM5: Biomarker Characterization and Fluorescent Probe Labeling for Image-Guided Gastric Cancer Surgery
by Serena Martinelli, Sara Peri, Cecilia Anceschi, Anna Laurenzana, Laura Fortuna, Tommaso Mello, Laura Naldi, Giada Marroncini, Jacopo Tricomi, Alessio Biagioni, Amedeo Amedei and Fabio Cianchi
Biomedicines 2025, 13(8), 1812; https://doi.org/10.3390/biomedicines13081812 - 24 Jul 2025
Viewed by 354
Abstract
Background: Gastric cancer (GC) is a malignant tumor of the gastrointestinal tract, characterized by high mortality rates and responsible for about one million new cases each year globally. Surgery is the main treatment, but achieving radical resection remains a relevant intraoperative challenge. [...] Read more.
Background: Gastric cancer (GC) is a malignant tumor of the gastrointestinal tract, characterized by high mortality rates and responsible for about one million new cases each year globally. Surgery is the main treatment, but achieving radical resection remains a relevant intraoperative challenge. Fluorescence-guided surgery offers clinicians greater capabilities for real-time detection of tumor nodules and visualization of tumor margins. In this field, the main challenge remains the development of fluorescent dyes that can selectively target tumor tissues. Methods: we examined the expression of the most suitable GC markers, including carcinoembryonic antigen cell adhesion molecule-5 (CEACAM5) and Claudin-4 (CLDN4), in GC cell lines. To further evaluate their expression, we performed immunohistochemistry (IHC) on tumor and healthy tissue samples from 30 GC patients who underwent partial gastrectomy at the Digestive System Surgery Unit, AOU Careggi, Florence. Additionally, we validated anti-CEACAM5 expression on patient-derived organoids. Furthermore, we developed a fluorescent molecule targeting CEACAM5 on the surface of GC cells and assessed its binding properties on patient tissue slices and fragments. Results: in this work, we first identified CEACAM5 as an optimal GC biomarker, and then we developed a fluorescent antibody specific for CEACAM5. We also evaluated its binding specificity for GC cell lines and patient-derived tumor tissue, achieving an optimal ability to discriminate tumor tissue from healthy mucosa. Conclusions: Overall, our results support the development of our fluorescent antibody as a promising tumor-specific imaging agent that, after further in vivo validation, could improve the accuracy of complete tumor resection. Full article
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23 pages, 4767 KiB  
Review
Self-Reporting H2S Donors: Integrating H2S Release with Real-Time Fluorescence Detection
by Changlei Zhu and John C. Lukesh
Chemistry 2025, 7(4), 116; https://doi.org/10.3390/chemistry7040116 - 21 Jul 2025
Viewed by 369
Abstract
Hydrogen sulfide (H2S), once regarded solely as a highly toxic gas, is now recognized as a crucial signaling molecule in plants, bacteria, and mammals. In humans, H2S signaling plays a role in numerous physiological and pathological processes, including vasodilation, [...] Read more.
Hydrogen sulfide (H2S), once regarded solely as a highly toxic gas, is now recognized as a crucial signaling molecule in plants, bacteria, and mammals. In humans, H2S signaling plays a role in numerous physiological and pathological processes, including vasodilation, neuromodulation, and cytoprotection. To exploit its biological functions and therapeutic potential, a wide range of H2S-releasing compounds, known as H2S donors, have been developed. These donors are designed to release H2S under physiological conditions in a controlled manner. Among them, self-reporting H2S donors are seen as a particularly innovative class, combining therapeutic delivery with real-time fluorescence-based detection. This dual functionality enables spatiotemporal monitoring of H2S release in biological environments, eliminating the need for additional sensors or probes that could disrupt cellular homeostasis. This review summarizes recent advancements in self-reporting H2S donor systems, organizing them based on their activation triggers, such as specific bioanalytes, enzymes, or external stimuli like light. The discussion covers their design strategies, performance in biological applications, and therapeutic potential. Key challenges are also highlighted, including the need for precise control of H2S release kinetics, accurate signal quantification, and improved biocompatibility. With continued refinement, self-reporting H2S donors offer great promise for creating multifunctional platforms that seamlessly integrate diagnostic imaging with therapeutic H2S delivery. Full article
(This article belongs to the Special Issue Organic Chalcogen Chemistry: Recent Advances)
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14 pages, 2816 KiB  
Article
A Colorimetric/Ratiometric Fluorescent Probe Based on Aggregation-Induced Emission Effect for Detecting Hypochlorous Acid in Real Samples and Bioimaging Applications
by Junliang Chen, Pingping Xiong, Huawei Niu, Weiwei Cao, Wenfen Zhang and Shusheng Zhang
Foods 2025, 14(14), 2491; https://doi.org/10.3390/foods14142491 - 16 Jul 2025
Viewed by 318
Abstract
Hypochlorous acid (HClO) serves as a biological mediator and is widely utilized as a disinfectant in food processing and water treatment. However, excessive HClO residues in food and environmental water raise concerns due to the potential formation of carcinogenic chlorinated byproducts and disinfection [...] Read more.
Hypochlorous acid (HClO) serves as a biological mediator and is widely utilized as a disinfectant in food processing and water treatment. However, excessive HClO residues in food and environmental water raise concerns due to the potential formation of carcinogenic chlorinated byproducts and disinfection byproducts (DBPs). Despite its importance, traditional methods for HClO detection often involve complex sample preparation, sophisticated instrumentation, and skilled operators. Herein, we report an aggregation-induced emission (AIE) small molecule fluorescent probe (NYV) that integrates colorimetric and ratiometric fluorescence responses for the detection of HClO. This probe exhibits high sensitivity, with a detection limit of 0.35 μM, a rapid response time of 1 min, and a wide linear range (0–142.5 μM), along with anti-interference capabilities, making it suitable for real-time monitoring. Furthermore, we have developed a portable solid-state sensor based on probe NYV for the rapid visual detection of HClO. The potential applications of this probe in real sample analysis and bioimaging experiments are demonstrated. Our findings contribute to the development of innovative fluorescent probes for HClO detection, with broad applications in food safety, environmental monitoring, and biomedical research on oxidative stress and ferroptosis. Full article
(This article belongs to the Section Food Analytical Methods)
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19 pages, 2749 KiB  
Article
Mechanism of Fluorescence Characteristics and Application of Zinc-Doped Carbon Dots Synthesized by Using Zinc Citrate Complexes as Precursors
by Yun Zhang, Yiwen Guo, Kaibo Sun, Xiaojing Li, Xiuhua Liu, Jinhua Zhu and Md. Zaved Hossain Khan
C 2025, 11(3), 48; https://doi.org/10.3390/c11030048 - 7 Jul 2025
Viewed by 494
Abstract
Zn-doped carbon dots (Zn@C-210 calcination temperature at 210 °C and Zn@C-260 calcination temperature at 260 °C) were synthesized via an in situ calcination method using zinc citrate complexes as precursors, aiming to investigate the mechanisms of their distinctive fluorescence properties. A range of [...] Read more.
Zn-doped carbon dots (Zn@C-210 calcination temperature at 210 °C and Zn@C-260 calcination temperature at 260 °C) were synthesized via an in situ calcination method using zinc citrate complexes as precursors, aiming to investigate the mechanisms of their distinctive fluorescence properties. A range of analytical methods were employed to characterize these nanomaterials. The mechanism study revealed that the coordination structure of Zn-O, formed through zinc doping, can induce a metal–ligand charge-transfer effect, which significantly increases the probability of radiative transitions between the excited and ground states, thereby enhancing the fluorescence intensity. The Zn@C-210 in a solid state and Zn@C-260 in water exhibited approximately 71.50% and 21.1% quantum yields, respectively. Both Zn@C-210 and Zn@C-260 exhibited excitation-independent luminescence, featuring a long fluorescence lifetime of 6.5 μs for Zn@C-210 and 6.2 μs for Zn@C-260. Impressively, zinc-doped CDs displayed exceptional biosafety, showing no acute toxicity even at 1000 mg/kg doses. Zn@C-210 has excellent fluorescence in a solid state, showing promise in anti-photobleaching applications; meanwhile, the dual functionality of Zn@C-260 makes it useful as a folate sensor and cellular imaging probe. These findings not only advance the fundamental understanding of metal-doped carbon dot photophysics but also provide practical guidelines for developing targeted biomedical nanomaterials through rational surface engineering and doping strategies. Full article
(This article belongs to the Special Issue Carbon Nanohybrids for Biomedical Applications (2nd Edition))
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21 pages, 7004 KiB  
Article
Mn-Doped Carbon Dots as Contrast Agents for Magnetic Resonance and Fluorescence Imaging
by Corneliu S. Stan, Adina Coroaba, Natalia Simionescu, Cristina M. Uritu, Dana Bejan, Laura E. Ursu, Andrei-Ioan Dascalu, Florica Doroftei, Marius Dobromir, Cristina Albu and Conchi O. Ania
Int. J. Mol. Sci. 2025, 26(13), 6293; https://doi.org/10.3390/ijms26136293 - 29 Jun 2025
Viewed by 647
Abstract
Carbon nanodots have recently attracted attention as fluorescence imaging probes and magnetic resonance imaging (MRI) contrast agents in diagnostic and therapeutic applications due to their unique optical properties. In this work we report the synthesis of biocompatible Mn (II)-doped carbon nanodots and their [...] Read more.
Carbon nanodots have recently attracted attention as fluorescence imaging probes and magnetic resonance imaging (MRI) contrast agents in diagnostic and therapeutic applications due to their unique optical properties. In this work we report the synthesis of biocompatible Mn (II)-doped carbon nanodots and their performance as fluorescence and MRI contrast agents in in vitro assays. The thermal decomposition of a Diphenylhydantoin–Mn(II) complex assured the incorporation of manganese (II) ions in the carbon dots. The obtained materials display a favorable spin density for MRI applications. The synthesized Mn(II)-CNDs also displayed remarkable photoluminescence, with a bright blue emission and good response in in vitro fluorescence imaging. Cytotoxicity investigations revealed good cell viability on malignant melanoma cell lines in a large concentration range. A cytotoxic effect was observed for MG-63 osteosarcoma and breast adenocarcinoma cell lines. The in vitro MRI assays demonstrated the potentialities of the Mn(II)-CNDs as T2 contrast agents at low dosages, with relaxivity values higher than those of commercial ones. Due to the simplicity of their synthetic pathway and their low cytotoxicity, the prepared Mn(II)-CNDs are potential alternatives to currently used contrast agents based on gadolinium complexes. Full article
(This article belongs to the Section Materials Science)
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19 pages, 5616 KiB  
Communication
A Poly(methacrolein-co-methacrylamide)-Based Template Anchoring Strategy for the Synthesis of Fluorescent Molecularly Imprinted Polymer Nanoparticles for Highly Selective Serotonin Sensing
by Madhav Biyani, Mizuki Matsumoto and Yasuo Yoshimi
Nanomaterials 2025, 15(13), 977; https://doi.org/10.3390/nano15130977 - 24 Jun 2025
Viewed by 420
Abstract
Neurotransmitters such as serotonin regulate key physiological and cognitive functions, yet real-time detection remains challenging due to the limitations of conventional techniques like amperometry and microdialysis. Fluorescent molecularly imprinted polymer nanoparticles (fMIP-NPs) offer a promising alternative and are typically synthesized via solid-phase synthesis, [...] Read more.
Neurotransmitters such as serotonin regulate key physiological and cognitive functions, yet real-time detection remains challenging due to the limitations of conventional techniques like amperometry and microdialysis. Fluorescent molecularly imprinted polymer nanoparticles (fMIP-NPs) offer a promising alternative and are typically synthesized via solid-phase synthesis, in which template molecules are covalently immobilized on a solid support to enable site-specific imprinting. However, strong template–template interactions during this process can compromise selectivity. To overcome this, we incorporated a poly(methacrolein-co-methacrylamide)-based template anchoring strategy to minimize undesired template interactions and enhance imprinting efficiency. We optimized the synthesis of poly(methacrolein-co-methacrylamide) under three different conditions by varying the monomer compositions and reaction parameters. The poly(methacrolein-co-methacrylamide) synthesized under Condition 3 (5:1 methacrolein-to-methacrylamide molar ratio, 1:150 initiator-to-total monomer ratio, and 4.59 M total monomer concentration) yielded the most selective fMIP-NPs, whose fluorescence intensity increased with an increase in serotonin concentration, rising by up to 37% upon serotonin binding. This improvement is attributed to higher aldehyde functionality in the poly(methacrolein-co-methacrylamide) which enhances template immobilization and generates a rigid imprinted cavity to interact with serotonin. These findings suggest that the developed fMIP-NPs hold significant potential as imaging probes for neurotransmitter detection, contributing to advanced studies in neural network analysis. Full article
(This article belongs to the Special Issue Recent Advances in the Development of Nano-Biomaterials)
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16 pages, 2051 KiB  
Article
Continuous Wavelet Transform-Based Method for High-Sensitivity Detection of Image Signals of Fluorescence Lateral Flow Assay
by Tao Zhang, Xiaosong Wu, Qian Wang, Long Zhang, Zhigang Li, Yangyang Peng, Qian Bian, Hui Shi, Yong Liu and Shu Wang
Sensors 2025, 25(13), 3846; https://doi.org/10.3390/s25133846 - 20 Jun 2025
Viewed by 402
Abstract
Fluorescence lateral flow assays (FLFA) based on quantum dot probes have attracted significant attention in recent years due to their high sensitivity and quantitative detection capabilities. FLFA requires the use of a straightforward fluorescence reader for quantitative detection. Most fluorescence readers employ narrowband [...] Read more.
Fluorescence lateral flow assays (FLFA) based on quantum dot probes have attracted significant attention in recent years due to their high sensitivity and quantitative detection capabilities. FLFA requires the use of a straightforward fluorescence reader for quantitative detection. Most fluorescence readers employ narrowband filters for auxiliary imaging, which facilitates the acquisition of high-contrast signals. However, during trace detection, the weak signal from FLFA can be easily lost due to optical flux loss associated with narrowband filters, thereby indirectly diminishing detection sensitivity. To address this issue, we developed a fluorescence signal reader that employs CMOS imaging without optical filters and proposed a highly sensitive signal detection algorithm based on continuous wavelet transform (CWT) to identify weak fluorescence signals with low contrast. Experimental results demonstrate that the method achieves a fluorescence detection sensitivity for quantum dots of 10−10 mol/L, with a relative standard deviation (RSD) of < 1.45%. The designed filter-free detection system and CWT analysis algorithm were applied to various FLFA systems (including the sandwich method and the competition method), with the correlation coefficient (R2) between all detection results and sample concentration exceeding 0.997. The findings of this study offer a highly sensitive signal detection method for the precise quantification of FLFA. Full article
(This article belongs to the Section Biomedical Sensors)
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15 pages, 27538 KiB  
Article
A Near-Infrared Fluorescent Probe for Specific Imaging of Lymph Node Metastases in Ovarian Cancer via Active Targeting of the Gonadotropin-Releasing Hormone Receptor
by Qiyu Liu, Jiaan Sun, Xiaobo Zhou, Mingxing Zhang, Tao Pu, Xiaolan Gao, Meng Zhang, Congjian Xu and Xiaoyan Zhang
Biomolecules 2025, 15(6), 868; https://doi.org/10.3390/biom15060868 - 14 Jun 2025
Viewed by 622
Abstract
Lymph node metastases are common in advanced ovarian cancer and are associated with poor prognosis. Accurate intraoperative identification of lymph node metastases remains a challenge in ovarian cancer surgery due to the lack of tumor-specific intraoperative imaging tools. Here, we developed a gonadotropin-releasing [...] Read more.
Lymph node metastases are common in advanced ovarian cancer and are associated with poor prognosis. Accurate intraoperative identification of lymph node metastases remains a challenge in ovarian cancer surgery due to the lack of tumor-specific intraoperative imaging tools. Here, we developed a gonadotropin-releasing hormone receptor (GnRHR)-targeted near-infrared (NIR) fluorescent probe, GnRHa-PEG-Rh760, through conjugation of a GnRH analog peptide with the Rh760 fluorophore and polyethylene glycol (PEG). A non-targeted probe (PEG-Rh760) served as control. In mouse models of subcutaneous xenografts, peritoneal and lymph node metastases derived from ovarian cancer cells, GnRHa-PEG-Rh760 showed superior tumor-specific accumulation. NIR fluorescence imaging revealed strong fluorescence signals localized to primary tumors, peritoneal lesions, and metastatic lymph nodes with no off-target signals in normal lymph nodes. The spatial co-localization between the NIR fluorescence of GnRHa-PEG-Rh760 and tumor-derived bioluminescence clearly confirmed the probe’s target specificity. GnRHa-PEG-Rh760 mainly accumulated in the tumor and liver and was gradually cleared at 96 h post-injection. The retention of fluorescence signals in normal ovary tissue further validated GnRHR-mediated binding of the probe. Notably, GnRHa-PEG-Rh760 exhibited excellent biocompatibility with no observed systemic toxicity as evidenced by hematologic and histopathologic analyses. These data demonstrate the potential of GnRHa-PEG-Rh760 as an intraoperative imaging agent, providing real-time fluorescence imaging guidance to optimize surgical precision. This study highlights the value of receptor-targeted molecular imaging probes in precision cancer surgery. Full article
(This article belongs to the Section Molecular Medicine)
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18 pages, 3449 KiB  
Article
Impact of Neoadjuvant Treatment on Target Expression in Rectal Cancer for Near-Infrared Tumor Imaging
by Elham Zonoobi, Lisanne K. A. Neijenhuis, Annelieke A. Lemij, Daan G. J. Linders, Ehsan Nazemalhosseini-Mojarad, Shadhvi S. Bhairosingh, N. Geeske Dekker-Ensink, Ronald L. P. van Vlierberghe, Koen C. M. J. Peeters, Fabian A. Holman, Rob A. E. M. Tollenaar, Denise E. Hilling, A. Stijn L. P. Crobach, Alexander L. Vahrmeijer and Peter J. K. Kuppen
Cancers 2025, 17(12), 1958; https://doi.org/10.3390/cancers17121958 - 12 Jun 2025
Viewed by 438
Abstract
Background: Rectal cancer (RC) patients with a clinical complete response (cCR) after neoadjuvant chemoradiotherapy (nCRT) may qualify for a watch-and-wait (W&W) approach. However, a 20–30% local tumor regrowth rate highlights challenges in identifying true responders. This study explores markers for future near-infrared fluorescence [...] Read more.
Background: Rectal cancer (RC) patients with a clinical complete response (cCR) after neoadjuvant chemoradiotherapy (nCRT) may qualify for a watch-and-wait (W&W) approach. However, a 20–30% local tumor regrowth rate highlights challenges in identifying true responders. This study explores markers for future near-infrared fluorescence tumor imaging by endoscopy to differentiate responders and the effect of nCRT on their expression. Methods: RC samples (n = 51) were collected from both pre-treatment biopsies and corresponding post-treatment surgical specimens. Samples were categorized by treatment response and determined using tumor regression grade (TRG) scoring. Immunohistochemistry assessed the expression of CEA, EpCAM, EGFR, and c-MET in tumors and adjacent normal tissues. Expression levels were quantified using H-scores (0–3), combining the percentage and intensity of stained cells. Pre- and post-treatment H-scores were compared to evaluate the impact of nCRT. Results: CEA, EpCAM, and c-MET were overexpressed in tumor tissue as compared to adjacent healthy mucosa in 100% (51/51), 98.4% (50/51), and 92% (47/51) of tumor biopsies, respectively, while EGFR showed no overexpression. A tumor-to-normal (T/N) ratio ≥ 2 was considered sufficient for differentiation in molecular fluorescence imaging. In pre-treatment biopsy samples, c-MET showed the highest T/N expression ratio (53% of the samples ≥ 2), followed by CEA (26.3%) and EpCAM (16%). Following nCRT treatment, CEA and c-MET maintained a ≥ 2 differential expression in 45% of all samples, whereas EpCAM exhibited this difference in only 9.2% of cases. Neoadjuvant therapy even significantly improved the T/N expression ratio for CEA and c-MET (p < 0.01) and EpCAM (p < 0.05), while EGFR expression remained lower than adjacent normal tissue. Significant increases in all marker expressions were observed in minimal responders (TRG4/5, p < 0.01–0.001), while near-complete responders (TRG2) exhibited non-significant changes in CEA, c-MET, and EGFR expression. Conclusions: c-MET and CEA emerged as optimal tumor imaging targets, showing sustained differential expression after nCRT. In vivo fluorescence-guided endoscopy using probes against these markers could play a role in future clinical decision-making. Full article
(This article belongs to the Special Issue Cancer Biomarkers—Detection and Evaluation of Response to Therapy)
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13 pages, 3184 KiB  
Article
Furin-Triggered Peptide Self-Assembly Activates Coumarin Excimer Fluorescence for Precision Live-Cell Imaging
by Peiyao Chen, Liling Meng, Yuting Wang, Xiaoya Yan, Meiqin Li, Yun Deng and Yao Sun
Molecules 2025, 30(11), 2465; https://doi.org/10.3390/molecules30112465 - 4 Jun 2025
Viewed by 609
Abstract
Monomer-to-excimer transition has become a valuable technique in fluorescence imaging because of its ability to enhance imaging contrast. However, from a practical perspective, the accuracy of excimer formation at target sites warrants further exploration. Enzyme-triggered peptide self-assembly provides a promising solution to this [...] Read more.
Monomer-to-excimer transition has become a valuable technique in fluorescence imaging because of its ability to enhance imaging contrast. However, from a practical perspective, the accuracy of excimer formation at target sites warrants further exploration. Enzyme-triggered peptide self-assembly provides a promising solution to this limitation. As a proof-of-concept, in this study, we developed a furin-triggered peptide self-assembling fluorescent probe RF-Cou by coupling a coumarin dye 7-(diethylamino)-2-oxo-2H-chromene-3-carboxylic acid (Cou) with a furin-responsive peptide scaffold for precision live-cell imaging. Upon entering furin-overexpressing 4T1 tumor cells, RF-Cou underwent enzymatic cleavage, releasing an amphiphilic peptide motif and self-assembling into nanoparticles largely concentrated in the Golgi apparatus to confine the diffusion of Cou. During this process, the Cou excimers were formed and induced a red shift in the fluorescence emission, validating the feasibility of RF-Cou in efficient excimer imaging of furin-overexpressing tumor cells. We expect that our findings will highlight the potential of stimuli-responsive small molecular peptide probes to advance excimer-based imaging platforms, particularly for enzyme-specific cell imaging and therapeutic monitoring. Full article
(This article belongs to the Special Issue Metal-Based Molecular Photosensitizers: From Design to Applications)
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