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过滤筛选
- 2017
- spectral reconstruction
- interference
- polarization
- transform
- imaging spectrometer
- Optoelectronic Information Science and Engineering
- BITTT
- Zhejiang University
- Guilin University of Aerospace Technology
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Cell Imaging Using Two-Photon Excited CdS Fluorescent Quantum Dots Working within the Biological Window
摘要: In recent years, two-photon excited semiconductor quantum dots (QDs) have been the subject of intense investigation due to their long excitation wavelength which helps to achieve deeper penetration and higher image resolution in optical bioimaging. In this paper, water-soluble CdS QDs were synthesized using a hydrothermal method and applied to human liver hepatocellular carcinoma (HepG2) cells. The first-principles calculation suggested that the S-rich defected structure contributes to a narrower band gap compared to the pristine structure. The resulting fluorescence wavelength was significantly red shifted, which was attributed to the deep defect states emission. The large Stokes shifts (> 200 nm) of the QDs can eliminate the possible cross-talk between the excitation light and the emission light. Two-photon induced red fluorescence emission can avoid overlapping with the autofluorescence emission of biological samples. The uptake and cell viability measurements of the HepG2 cells showed a good biocompatibility and a low toxicity of CdS QDs. Two-photon excited scanning microscopy images revealed that the HepG2 cells incubated with CdS QDs emitted bright red upconversion fluorescence and the fluorescence brightness was 38.2 times of that of the control group. These results support CdS QDs as a good candidate for application in cellular imaging.
关键词: two-photon absorption,CdS quantum dots,deep defect states,HePG2 cells,biological imaging
更新于2025-11-21 11:24:58
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Assessing retinal ganglion cell death and neuroprotective agents using real time imaging
摘要: The evaluation of retinal ganglion cell (RGC) death is a key part of retinal disease care. Previously, we used a Sytox Orange (SO)-based real-time imaging method to assess the RGCs in mice that underwent optic nerve crush. Here, we used N-methyl-D-aspartate (NMDA) injury in rats to confirm our model and assess the effect of neuroprotective agents on RGCs. The rats received NMDA injury and the intravitreal injection of SO, a cell-impermeant dyeing compound that targets nucleic acid. After ten minutes, non-invasive confocal scanning laser ophthalmoscopy visualized damaged or dying cells. Finally, the retinas were flat-mounted for histological confirmation of RGC death, with retrograde Fluorogold labeling and Alexa Fluor 488 Annexin V-conjugate (Annexin V) staining. This also revealed the time course of retinal cell death and the neuroprotective effect of SNJ-1945. Real-time imaging showed that SO-positive cells significantly increased starting 2 hours after NMDA injection and reached an approximate plateau at 3 hours. SO-positive cells were positive for Fluorogold and Annexin V in the isolated retinas. Moreover, the number of SO-positive retinal cells was significantly lower after treatment with SNJ-1945, compared to carboxymethyl cellulose. These results were confirmed in the isolated retinas. Thus, real-time imaging with SO allows the quick quantification of NMDA-induced RGC damage and death, and evaluation of neuroprotective agents. This technique may aid research into the development of new neuroprotective therapies.
关键词: retinal ganglion cell,Real-time imaging,SYTOX orange,neuroprotection
更新于2025-11-21 11:24:58
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Zwitterionic near-infrared fluorophore-conjugated epidermal growth factor for fast, real-time, and target-cell-specific cancer imaging
摘要: Epidermal growth factor receptor (EGFR) is overexpressed in many types of cancers, which is associated with metastatic potential and poor prognosis in cancer patients. Therefore, development of EGFR-targeted sensitive imaging probes has been a challenge in tumor targeting, image-guided cancer surgery, patient-selective anti-EGFR therapy, and efficient targeted therapies. Methods: We synthesized a zwitterionic near-infrared fluorophore (ATTO655)-conjugated epidermal growth factor (EGF) as a novel activatable molecular probe. Fluorescence OFF/ON property and EGFR-targeting specificity of EGF-ATTO655 as well as its utility in real-time near-infrared (NIR) fluorescence imaging of EGFR-positive cancers were evaluated using in vitro and in vivo studies. Results: When conjugated to EGF, the fluorescence of ATTO655 quenched efficiently by photo-induced electron transfer (PET) mechanism between the conjugated dyes and nearby amino acid quenchers (tryptophan/tyrosine residues), which was stably maintained at physiological pH and in the presence of serum for at least 17 h. The fluorescence of EGF-ATTO655 turned on by receptor-mediated endocytosis and subsequent disintegration of EGF in EGFR-positive A431 cancer cells, thereby enabling specific and real-time fluorescence imaging of EGFR-positive cancer cells. Consequently, EGFR-positive tumors could be clearly visualized 3 h post-injection with a significantly high tumor-to-background ratio (TBR = 6.37). Conclusion: This PET mechanism-based OFF/ON type of EGF probe showed great potential for rapid, real-time, and target-cell-specific imaging of EGFR-overexpressing cancers in vitro and in vivo.
关键词: photo-induced electron transfer,Epidermal growth factor,real-time cancer imaging,tumor-specific targeting
更新于2025-11-21 11:24:58
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Discovery of Turn-On Fluorescent Probes for Detecting Bcl-2 Protein
摘要: Bcl-2 (B cell lymphoma-2 gene) family proteins play a central role in regulating programmed cell death. In cancer, anti-apoptotic Bcl-2 proteins, such as Bcl-2 and Mcl-1, are overexpressed. However, there are few developed labeling techniques for tracing the dynamic processes of Bcl-2. To study the physiological process of Bcl-2 protein, a novel series of small molecule fluorescent probes (1-3) were designed and evaluated for their labeling properties. It’s interesting that our probes can be applied to identify tumor tissue slices and differentiate the tumor and normal tissues effectively, a feature that renders these probes compatible for future cancer diagnosis in clinical practice.
关键词: fluorescent probes,cell imaging,Bcl-2 protein
更新于2025-11-21 11:24:58
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Aggregation-Induced Emission: Lighting Up hERG Potassium Channel
摘要: Based on the scaffold of astemizole and E-4031, four AIE light-up probes (L1–L4) for Human Ether-a-go-go-Related Gene (hERG) potassium channel were developed herein using AIE fluorogen(TPE). These probes showing advantages such as low background interference, superior photostability, acceptable cell toxicity, and potent inhibitory activity, which could be used to image hERG channels at the nanomolar level. These AIE light-up probes hoped to provide guidelines for the design of more advanced AIE sensing and imaging hERG channels to a broad range of applications.
关键词: hERG channel,pharmacophore,fluorophore,cell imaging,AIE light-up probes
更新于2025-11-21 11:24:58
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Design and synthesis of a new fluorescent probe for cascade detection of Zn <sup>2+</sup> and H <sub/>2</sub> PO <sub/>4</sub><sup>?</sup> in water and targeted imaging of living cells
摘要: Design and synthesis of new fluorescence probes with good water‐solubility is of great importance to better understanding the significant role of ions which are related to biology and the environment. As important ions, zinc ion (Zn2+) and dihydrogen phosphate ion (H2PO4?) display essential roles in living systems, and quantitative detection of these ions in water is still a challenge. In order to consider the significant role of the galactose moiety in the design of a water‐soluble fluorescence sensor, herein, we have developed a novel probe, Gal‐AQTF, for the cascade detection of Zn2+ and H2PO4? with excellent selectivity in water. Through the introduction of the galactose moiety onto the sensor AQTF, which has been reported earlier by us, the water‐solubility, cell compatibility and targeting ability were enhanced. Gal‐AQTF has been successfully applied in the imaging of the living cells of HepG2 and A549, and illustrated good selectivity for the HepG2 cells which overly express the asialoglycoprotein (ASGP) receptor.
关键词: targeted imaging,Galactose,Zn2+ and H2PO4?,in water,fluorescent probe
更新于2025-11-21 11:24:58
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MnO2 Nanosheet-mediated Ratiometric Fluorescence Biosensor for MicroRNA Detection and Imaging in Living Cells
摘要: MicroRNA (miRNA) plays significant roles in cell proliferation, differentiation and apoptosis, and has been considered to be valuable biomarker for cancer. Accurate and sensitive detection of miRNA is crucially significant for cancer diagnosis and treatment. Here, a MnO2 nanosheet-mediated ratiometric fluorescence biosensor was designed for miRNA detection and imaging in living cells. It contained MnO2 nanosheets acting as DNA carrier, and fluorescent donor (FAM)-labeled hairpin H1 (recognition probe) and fluorescent acceptor (TAMRA)-labeled hairpin H2 (amplification probe). When the biosensor entered cell by endocytosis, MnO2 nanosheets were degraded to Mn2+ via intracellular glutathione (GSH) and the adsorbed hairpins H1 and H2 were released. The intracellular target miRNA-21 hybridized with the recognition unit of H1 to initiate catalyzed hairpin assembly (CHA) and a large amount of H1-H2 duplexes were produced. This brought fluorescent donor FAM and fluorescent acceptor TAMRA into close proximity to produce fluorescence resonance energy transfer (FRET), inducing a ratiometric fluorescent response (donor signal decreased and acceptor signal enhanced) for miRNA-21 detection. Furthermore, this method could be applied to differentiate the expression levels of miRNA-21 in HeLa, HepG-2 and L02 cells. These results indicated that the proposed method possessed great potential in the early diagnosis of miRNA-related diseases.
关键词: MicroRNA detection,MnO2 nanosheets,Ratiometric,Cell imaging
更新于2025-11-21 11:24:58
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Gaussian FRET two-hybrid assays for determining the stoichiometry of hetero-oligomeric complexes in single living cells
摘要: Here we integrate multiple Gaussian-functions analysis into fluorescence resonance energy transfer (FRET) two-hybrid assays (Gaussian FRET two-hybrid assay) to determine the stoichiometric ratios of intracellular hetero-oligomers in single living cells. This method adopts in multiple Gaussian-functions to fit the E-count histograms of both donor- and acceptor-centric FRET efficiency (ED and EA) images of a single cell for obtaining the peak values (EDi and EAi), thus yielding the corresponding stoichiometric ratios (EDi/EAi) of intracellular hetero-oligomers. We performed Gaussian FRET two-hybrid assay for living Hela cells coexpressing different FRET tandem plasmids, and obtained consistent results with the expected values. Gaussian FRET two-hybrid assay for cells coexpressing Bad-CFP and Bcl-XL-YFP reveals that Bcl-XL binds with Bad to form a hetero-oligomeric complex with a stoichiometry of 2:1 on mitochondria.
关键词: Multiple Gaussian-functions analysis,FRET imaging,Single living cell,Stoichiometry,Hetero-oligomeric complex
更新于2025-11-21 11:24:58
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Graphene-Based Steganographicly Aptasensing System for Information Computing, Encryption and Hiding, Fluorescent Sensing and In Vivo Imaging of Fish Pathogens
摘要: Inspired by information processing and communication of life based on complex molecular interactions, some artificial (bio)chemical systems have been developed for applications in molecular information processing or chemo/biosensing and imaging. However, little attention has been paid to simultaneously and comprehensively utilize the information computing, encoding and molecular recognition capabilities of molecular-level systems (such as DNA-based systems) for multifunctional applications. Herein, a graphene-based steganographicly aptasensing system was constructed for multifunctional application, which relies on specific molecular recognition and information encoding abilities of DNA aptamers (Aeromonas hydrophila and Edwardsiella tarda-binding aptamers as models) and the selective adsorption and fluorescence quenching capacities of graphene oxide (GO). Although graphene-DNA systems have been widely used in biosensors and diagnostics, our proposed graphene-based aptasensing system can not only be utilized for fluorescent sensing and in vivo imaging of fish pathogens (Aeromonas hydrophila and Edwardsiella tarda), but can also function as a molecular-level logic computing system where the combination of matters (specific molecules or materials) as inputs produces the resulting product (matter level) or fluorescence (energy level) changes as two outputs. More importantly and interestingly, our graphene-based steganographicly aptasensing system can also be served as a generally doubly cryptographic and steganographic system for sending different secret messages by using pathogen-binding DNA aptamers as information carriers, GO as a cover, a pair of keys: target pathogen as a public key, the encryption key used to encode or decode a message in DNA as a private key. Our study not only provides a novel nano-biosensing assay for rapid and effective sensing and in vivo imaging fish pathogens, but also demonstrates a prototype of (bio)molecular steganography as an important and interesting extension direction of molecular information technology, which is helpful in probably promoting the development of multifunctional molecular-level devices or machines.
关键词: aptasensing,steganography,graphene oxide,DNA aptamer,encryption,fish pathogens,in vivo imaging,information hiding
更新于2025-11-21 11:24:58
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Perylenequinone-based “turn on” fluorescent probe for hydrogen sulfide with a high sensitivity in living cells
摘要: Hydrogen sulfide (H2S) is a kind of gaseous signal molecule in many physiological processes. In order to detect H2S, a novel “turn on” fluorescent probe 6,12-dihydroxyperylene-1,7-dione (DPD) was designed and synthesized. The probe DPD is fluorescence silence, while the addition of H2S induces an obvious green fluorescence with an obvious color change from dark blue to yellow-green. The probe shows excellent selectivity, fast response (2.5 minutes) and linear curve (0-90 μM) in wide effective pH range (4-10). Competition experiments are also revealed in corresponding studies and the detection limit is 3.6 μM. The response mechanism is proved to be the reduction of the probe by H2S, which is confirmed by 1H NMR. Furthermore, through the fluorescence turn-on signal toward H2S in Hela cells, probe DPD was successfully applied to monitor H2S in living Hela cells.
关键词: hydrogen sulfide,probe,fluorescence imaging,cell imaging,perylenequinone
更新于2025-11-21 11:08:12