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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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[Methods in Molecular Biology] Plant Long Non-Coding RNAs Volume 1933 (Methods and Protocols) || Trimolecular Fluorescence Complementation (TriFC) Assay for Visualization of RNA-Protein Interaction in Plants
摘要： RNA-protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA-protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with the MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) to tag lncRNA. Target RNA is tagged with 6xMS2, and MCP and RNA-binding protein are fused with YFP fragments. DNA constructs encoding such fusion RNA and proteins are infiltrated into tobacco leaves with Agrobacterium suspensions. RNA-protein interaction in vivo is observed by confocal microscopy.
关键词： RNA-protein interaction,Long noncoding RNA,TriFC,Tobacco transient expression,In vivo visualization
更新于2025-11-21 11:08:12
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Correlated Light-Serial Scanning Electron Microscopy (CoLSSEM) for ultrastructural visualization of single neurons in vivo
摘要： A challenging aspect of neuroscience revolves around mapping the synaptic connections within neural circuits (connectomics) over scales spanning several orders of magnitude (nanometers to meters). Despite significant improvements in serial section electron microscopy (SSEM) technologies, several major roadblocks have impaired its general applicability to mammalian neural circuits. In the present study, we introduce a new approach that circumvents some of these roadblocks by adapting a genetically-encoded ascorbate peroxidase (APEX2) as a fusion protein to a membrane-targeted fluorescent reporter (CAAX-Venus), and introduce it in single pyramidal neurons in vivo using extremely sparse in utero cortical electroporation. This approach allows us to perform Correlated Light-SSEM (CoLSSEM), a variant of Correlated Light-EM (CLEM), on individual neurons, reconstructing their dendritic and axonal arborization in a targeted way via combination of high-resolution confocal microscopy, and subsequent imaging of its ultrastructural features and synaptic connections with ATUM-SEM (automated tape-collecting ultramicrotome - scanning electron microscopy) technology. Our method significantly will improve the feasibility of large-scale reconstructions of neurons within a circuit, and permits the description of some ultrastructural features of identified neurons with their functional and/or structural connectivity, one of the main goal of connectomics.
关键词： connectomics,APEX2,in vivo,Correlated Light-SSEM,single neurons,ultrastructural visualization
更新于2025-11-21 11:01:37
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Enhanced In‐vivo Optical Imaging of the Inflammatory Response to Acute Liver Injury in C57Bl/6 Mice using a Highly Bright Near‐Infrared BODIPY Dye
摘要： Delving deeper is possible in whole body in vivo imaging using a super-bright membrane targeting BODIPY dye (BD). The dye was employed to monitor homing of ex vivo, fluorescently labelled neutrophils to an injured liver of dark pigmented C57BL/6 mice. In Vivo Imaging System (IVIS) data conclusively showed an enhanced signal intensity and a higher signal-to-noise ratio in mice receiving neutrophils labelled with the BD dye compared to those labelled with a gold standard dye at 2 hr post in vivo administration of fluorescently labelled cells. Fluorescence-activated cell sorting (FACS) confirmed that BD was non-toxic, and an exceptional cell labelling dye that opens up precision deep organ in vivo imaging of inflammation in mice routinely used for biomedical research. The origin of enhanced performance is identified with the molecular structure, and the distinct localisation of the dye within cells that enable remarkable changes in its optical parameters.
关键词： In-vivo imaging,Cell Sorting,Bodipy,Liver,Fluorescence
更新于2025-11-19 16:46:39
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A near-infrared fluorescence probe for ultrafast and selective detection of peroxynitrite with large Stokes shift in inflamed mouse models
摘要： Peroxynitrite (ONOO?) is a kind of reactive oxygen species (ROS) which is associated with pathogenesis of many diseases. A new near-infrared fluorescence probe (DCM-OH) which based on dicyanomethylene-4H-pyrans to detect endogenous ONOO? was designed and synthesized. The two-photon absorption cross sections and large Stokes shift make the probe deeper issue penetration and lower self-absorption. The obtained results demonstrated that probe DCM-OH could sensitively detect ONOO? with a low detection limit (53 nM). What’s more, probe DCM-OH exhibited an ultrafast response rate (within 5 s) toward ONOO?, which would be in favor of tracking the highly reactive and short-lived ONOO? in the living systems. Moreover, DCM-OH was successfully employed for imaging endogenous ONOO? in HepG2/RAW 264.7 cells and further applied to visualize oxidative stress in mouse model of inflammation.
关键词： In vivo imaging,NIR,Fluorescence probe,Peroxynitrite,Large Stokes shift
更新于2025-11-14 15:29:11
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Synthesis, In Vitro and In Vivo Behaviour of TiO2 Containing Inorganic/Organic Hybrids
摘要： In the present study inorganic/organic hybrids reinforced by introducing titanium (TiO2) in the form of Ti-n-butoxide were prepared. The phase composition, microstructure and mechanical properties of the prepared hybrids were investigated. Increasing the titanium content gradually enhanced the mechanical data of the prepared hybrids. The formation of apatite on the surface of the prepared hybrids was examined in SBF (simulated body fluid). In vivo studies revealed the ability of the hybrids to regenerate bone tissue in femur defects of adult male rabbits five months after surgery. The prepared hybrids are considered to be promising materials for bone substitutes or bone filler.
关键词： in vitro test,in vivo test,mechanical properties,inorganic/organic hybrids,microstructure
更新于2025-09-23 15:23:52
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Progrès dans les méthodes diagnostiques du déficit en cellules souches limbiques. Apport de la microscopie confocale et de la tomographie en cohérence optique
摘要： The limbus is the anatomical and functional barrier between corneal and conjunctival epithelia. It is characterized by presence of the limbal stem cell niche which allows corneal homeostasis to be maintained. Limbal stem cell deficiency is characterized by a dual process: insufficient regeneration of corneal epithelium, which cannot therefore assure its function of physiological support, associated with corneal invasion by conjunctival proliferation. Diagnosis is currently made via routine clinical examination, corneal impression cytology and in vivo confocal microscopy (IVCM). Slit lamp examination shows abnormal limbal anatomy, thin and irregular epithelium with late fluorescein staining, and superficial vascularization. With its high resolution, IVCM allows identification of limbal and corneal epithelial changes at a cellular level in en face views, parallel to the corneal surface, but with a restricted viewing field of the corneal surface. It shows a poor transition between the corneal and conjunctival epithelia, associated with a loss of the normal corneal epithelial stratification, low basal cell and sub-basal nerve plexus densities, even with sub-epithelial fibrosis. Optical coherence tomography in central cornea and at the limbus, with scans in different orientations, allows a quick, global and non-invasive analysis of normal eyes and those with limbal stem cell deficiency. It shows a thin limbal epithelium, lacking normal thickening, featuring absence of stromal undulations and limbal crypts in cross-sections and sections parallel to the limbus, lack of visible limbal crypts in en face sections, loss of clear transition between the hyporeflective corneal epithelium and the hyperreflective conjunctival epithelium, and hyperreflective sub-epithelial fibrosis.
关键词： Limbal stem cell deficiency,Limbus,Optical coherence tomography,In vivo confocal microscopy
更新于2025-09-23 15:23:52
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Radioiodinated Portable Albumin Binder as a Versatile Agent for in vivo Imaging with Single-Photon Emission Computed Tomography
摘要： In this study, radioiodinated 4-(p-iodophenyl)butyric acid ([131I]IBA) was synthesized and evaluated as a portable albumin-binder for potential applications in SPECT imaging of blood pool, tumor and lymph node with significantly improved pharmacokinetic properties. The [131I]IBA was prepared under the catalyst of Cu2O/1,10-phenanthroline. After that, the albumin-binding capability of [131I]IBA was tested in vitro, ex vivo and in vivo, respectively. [131I]IBA was obtained with very high radiolabeling yield (> 99%) and good radiochemical purity (> 98%) within 10 min. It binds to albumin effectively with high affinity (IC50= 46.5 μM) and has good stability. The results of biodistribution indicated that the [131I]IBA was mainly accumulated in blood with good retention (10.51±2.58%ID/g at 30 min p.i. and 4.63±0.17%ID/g at 4 h p.i.). In the SPECT imaging of mice models with [131I]IBA, blood pool, lymph node and tumors could be imaged clearly with high target-to-background ratio. Overall, the radioiodinated albumin binder of [131I]IBA with long blood half-life and excellent stability could be used to decorate diversified albumin-binding radioligands and developed as a versatile theranostic agent.
关键词： SPECT,in vivo imaging,radioiodination,albumin binder,animal models
更新于2025-09-23 15:23:52
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Zn <sub/>3</sub> Ga <sub/>2</sub> Ge <sub/>2</sub> O <sub/>10</sub> :Cr <sup>3+</sup> Uniform Microspheres: Template-Free Synthesis, Tunable Bandgap/Trap Depth, and <i>In Vivo</i> Rechargeable Near-Infrared-Persistent Luminescence
摘要： Near-infrared (NIR) emitting persistent phosphors of Cr3+-doped zinc gallogermanate have emerged for in vivo bio-imaging with the advantages of no need for in situ excitation. However, it is challenging to synthesize well-dispersed and uniform spherical particles with high brightness, high resolution, and distinguished NIR long afterglow. In this work, Zn3Ga2Ge2O10:Cr3+ (ZGGC) monospheres were directly synthesized by a facile hydrothermal method with the assistance of citric anions (Cit3-), which emit a NIR emission at ~696 nm and exhibit excellent NIR persistent luminescence with rechargeability. Controlled experiments indicated that the shape evolution of ZGGC product is significantly affected by Cit3-, solution pH, and the duration and temperature of hydrothermal reaction. Furthermore, compositional influence on the crystal structure, bandgap, trap depth, and luminescence characteristics of ZnyGa2Ge2O10-δ:Cr3+ (y = 2.8, 3.0, 3.2) were investigated in details, which allows to construct an energy level diagram of the ZGGC host, Cr3+ ions, and electron traps. It was found that the bandgap and conduction-band minimum (CBM) are significantly affected by the Zn content, while the valence-band maximum (VBM) is not. The y = 3.0 sample exhibited the best persistent luminescence, owing to its deepest defects. The ZGGC-NH2 prepared through surface functionalization of ZGGC spheres showed distinguished NIR long afterglow, low toxicity, and great potential for in vitro cell imaging and in vivo bio-imaging in the absence of excitation. Moreover, the persistent-luminescence signal from the ZGGC-NH2 can be repeated in vivo through in situ recharge with external excitation of a red LED lamp, indicating that the ZGGC-NH2 is suitable for applications in long-term in vivo imaging.
关键词： in vivo imaging,Near infrared persistent luminescence,conduction band minimum,monospheres
更新于2025-09-23 15:23:52
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Core–Satellite Nanomedicines for <i>in Vivo</i> Real-Time Monitoring of Enzyme-Activatable Drug Release by Fluorescence and Photoacoustic Dual-Modal Imaging
摘要： It remains an unresolved challenge to achieve spatial and temporal monitoring of drug release from nanomedicines (NMs) in vivo, which is of crucial importance in disease treatment. To tackle this issue, we constructed core?satellite ICG/DOX@Gel-CuS NMs, which consist of gelatin (Gel) nanoparticles (NPs) with payloads of near-infrared fluorochrome indocyanine green (ICG) and chemo-drug doxorubicin (DOX) and surrounding CuS NPs. The fluorescence of ICG was initially shielded by satellite CuS NPs within the intact ICG/DOX@Gel-CuS NMs and increased in proportion to the amount of DOX released from NMs in response to enzyme-activated NMs degradation. For more comprehensive understanding of the drug-release profile, a theoretical model derived from computer simulation was employed to reconstruct the enzyme-activatable drug release of the ICG/DOX@Gel-CuS NMs, which demonstrated the underlying kinetics functional relationship between the released DOX amount and recovered ICG fluorescence intensity. The kinetics of drug release in vivo was assessed by administrating ICG/DOX@Gel-CuS NMs both locally and systemically into MDA-MB-231 tumor-bearing mice. Upon accumulation of ICG/DOX@Gel-CuS NMs in the tumor, overexpressed enzymes triggered the degradation of the gelatin scaffold as well as the release of DOX and ICG, which can be visually depicted with the ICG fluorescence signal increasing only in the tumor area by fluorescence imaging. Additionally, the photoacoustic signal from CuS NPs was independent from the physical status of ICG/DOX@Gel-CuS NMs and hence was utilized for real-time NMs tracking. Thus, by taking advantage of the core?satellite architecture and NMs degradability in tumor site, the DOX release profile of ICG/DOX@Gel-CuS NMs was monitored by fluorescence and photoacoustic dual-modal imaging in a real-time noninvasive manner.
关键词： core?satellite,nanomedicines,drug release in vivo,dual-modal imaging,computer simulation
更新于2025-09-23 15:23:52