45 results about "Live cell imaging" patented technology

The invention discloses a functionalized targeting fluorescent nanoprobe and application thereof in protein mutation analysis, and relates to the technical field of biology, the fluorescent nanoprobe comprises a targeting molecule-PEG hydrophilic bridging molecule and a matrix; the preparation method comprises the following steps: adding nano-seeds into fluorescent molecules to prepare fluorescent nano-particles, and stirring and mixing the fluorescent nano-particles with targeting molecules-PEG hydrophilic bridging molecules; the method is combined with deep learning to be applied to protein mutation analysis. The multifunctional integrated protein mutation receptor targeted fluorescent nanoprobe provided by the invention can accurately mark the position of a mutant protein at a single cell level, and can realize in-situ, real-time and dynamic living cell imaging of a tumor cell endogenous molecular mutant protein; and qualitative and quantitative analysis of tumor cell protein mutation can be realized through subcellular localization of the targeted fluorescent nanoprobe.

The invention discloses a targeted KRAS G12C environment-sensitive fluorescent probe as well as a preparation method and application thereof, the fluorescent probe is a compound shown in the following formula or pharmaceutically acceptable salt thereof, and the structural formula is shown in the specification. The invention also specifically discloses a preparation process of the targeted KRAS G12C environment-sensitive fluorescent probe and application of the targeted KRAS G12C environment-sensitive fluorescent probe in preparation of a living cell imaging detection preparation. According to the invention, a known KRAS G12C covalent inhibitor structure is reasonably modified according to a molecular docking mode of the inhibitor and KRAS G12C, and an environment-sensitive fluorescent labeling group is introduced to form the environment-sensitive fluorescent probe with a switch mechanism, so that the environment-sensitive fluorescent probe has the advantages of simplicity and convenience in operation, no need of washing, no need of paraformaldehyde fixation of cells and the like; the invention aims to realize the intuitive visualization of KRAS G12C in living cells / in vivo, and assists in the research and visual application of targeted therapy.

The present technique provides a simple, cost-effective and robust method for re-alignment of interfaces between cell conjugates parallel to the coverslip. The technique is based on placement of two cell types, to subsequently form an interaction between the cell types, to opposing coverslips and then bringing them together before or during imaging, for either fixed or live cell imaging. Spacer particles having defined parameters control the z-separation and the relative lateral position of the opposing coverslips. We show that our method allows most types of super-resolution imaging.

The utility model relates to a cooling assembly for a living cell imager, which comprises a base and a water storage tank, the base is provided with a cooling water inlet and a cooling water return port, a water flow channel is formed between the cooling water inlet and the cooling water return port, the water storage tank is arranged below the water flow channel, and the water storage tank is arranged below the cooling water inlet and the cooling water return port. And the water storage tank is provided with a condensate water outflow channel. The water storage tank is additionally arranged in the water cooling system of the living cell imager, so that condensed water generated in the water cooling process can be stored, and then the condensed water with the temperature consistent with that in the incubator is discharged into the incubator. The cooling requirement of the living cell imager is met, and the temperature in the incubator is not changed. The problem of how to enable the living cell imager to normally work in the incubator for a long time is effectively solved.

The invention discloses an automatic focusing module and an automatic focusing method for high-flux living cell imaging. The automatic focusing module comprises a base; a Z-axis moving mechanism is arranged on the base; a converter fixing seat is arranged on the Z-axis moving mechanism; an electric converter is arranged on the converter fixing seat; the electric converter is provided with a laser automatic focusing sensor and an objective lens. The laser automatic focusing sensor and the objective lens are integrated on the electric converter, when the laser automatic focusing sensor detects the distance of a specimen, the laser automatic focusing sensor does not need to move along the X-axis and the Y-axis, the position of the objective lens only needs to be switched through the electric converter, the Z-axis moving mechanism drives the objective lens to move to a proper position along the Z-axis, and then photographing and imaging can be completed. Errors caused by repeated displacement can be effectively avoided, the imaging effect is improved, the working efficiency can be improved, and the working time can be shortened.

The invention discloses an antibody targeting light activated fluorescent protein Dronpa and related application thereof.The amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 in a heavy chain variable region of the antibody are shown as SEQ ID NO.4-6 respectively, and the amino acid sequences of CDR-L1, CDR-L2 and CDR-L3 in a light chain variable region of the antibody are shown as SEQ ID NO.7-9 respectively. According to the antibody and the application thereof, accurate tool support is provided for detection and analysis of Dronpa protein, and the antibody can be applied to detection of Dronpa protein. Meanwhile, a key technical guarantee is provided for life science experiments (such as living cell imaging and molecular mechanism research) and potential clinical application based on the Dronpa protein.

The present application belongs to the field of chemical synthesis and biotechnology, and particularly relates to a kind of sugar metabolism product molecular probes and preparation method thereof. Starting from fructose, a bio-orthogonal group is added to the 4-position hydroxyl group of the compound without affecting the biological activity of the sugar metabolism product. The present application provides a kind of sugar metabolism product probe and preparation method thereof, compared with the prior art, has the following beneficial effects: (1) the present application first synthesizes an endogenous sugar metabolism product molecular probe that can undergo click chemistry reaction, by adding a bio-orthogonal group to the 4-position hydroxyl group, the biological activity of the sugar metabolism product is maintained, which can be used for live cell imaging; (2) the probe of the present application has not been reported, and the synthesis method has less side reaction, mild reaction condition, good selectivity, can reflect the behavior of endogenous sugar metabolism product in cells, including direct reaction of sugar metabolism product in live cells Subcellular localization and tracing, is an important technical support for the study of live cell sugar metabolism.

The present invention relates to novel compounds capable of localized fluorescent signal amplification by peroxidase, a method for preparing them, and their uses as fluorescent probes. The compounds according to the present invention serve as new fluorescent amplification probes utilizing peroxidase, enabling the generation of a localized labeling pattern at the target site. Through this, they provide super-resolution protein localization information and allow the visualization of interactions between various organelles. Furthermore, they can provide localized and stable fluorescent signals in CLEM and super-resolution imaging and enable selective fluorescent labeling under fixed conditions, making them highly applicable in future proteomics and live-cell imaging.

Provided herein are methods for analysis of target cells on a population or individual basis, including before and after contact with a stimulus in order to determine the effect of such stimulus on the target cells. Also provided are devices for performing such methods. The analysis methods involve identifying and measuring or tracking morphological changes that occur in target cells over a period of time. Tracking is accomplished using imaging systems capable of imaging target cells individually over a period of time either continuously or at discrete intervals of time.

The invention relates to the technical field of chemical genetics marking, in particular to a marking probe for quantifying protein dynamics. The probe comprises a compound with the following structural formula, wherein the compound contains a halogenated alkane chain and can be specifically covalently bound with halogenated alkane dehalogenase-HaloTag tag protein. The HaloTag-containing fusion target protein is combined with the HaloTag-containing fusion target protein in cells, and has the advantages of high specificity and stability, low toxicity, good biocompatibility and strong photobleaching resistance in living cell imaging. The compound has the property of space-limited emission effect, and can sensitively mark and respond to the spatial dynamic change of any single protein molecule in the physiological process of cells through a HaloTag bridge, so that the dynamic quantification and visualization of the protein are realized in the method, and the compound is used for recording physiological activities under different time scales.

Aspects of the application provide methods and compositions for identifying and evaluating putative therapeutic agents for cancer by live cell imaging. Cell samples comprising cancerous cells that have been pre-treated with a test agent are contacted with a BH3 peptide, and samples are imaged by live cell imaging over a time interval. Methods of the application can be used to determine whether a patient is likely to benefit from treatment with a particular test agent.

The invention relates to the technical field of biology, and discloses a recovery liquid for improving recovery efficiency and vitality of organoids, which comprises a metal ion chelating agent, a dissociation agent and a cell protective agent. According to the recovery liquid, matrigel is mildly disintegrated through cooperation of the metal ion chelating agent and the dissociation agent, the organoid can be separated from the matrix within 30 minutes, and the recovery rate of the organoid reaches 90% or above and is far higher than that of a pure mechanical blowing method; the dissociation agent is used for replacing pancreatin and other strong digestive enzymes, and is matched with the cell protective agent, so that the survival rate of the recovered organoid is greater than or equal to 95%, the three-dimensional structure and the intercellular connection are kept complete, and the biological function is real; all the components are clear, the batch is stable, the operation is simple and convenient, the storage is convenient, the culture medium is suitable for passage, collection, downstream cell counting, molecular detection, immunofluorescence, living cell imaging and the like of various sources of organoids such as gastrointestinal tracts, livers and lungs, and the standardization and repeatability of organoid culture are remarkably improved.

The utility model discloses a light-emitting diode (LED) array light source light following system of a high-flux living cell imager, which relates to the technical field of instrument design and control and comprises an objective table and an LED light source which are relatively fixed, an objective lens moves relative to the objective table, the movement stroke of the objective lens is matched with the LED light source, and the light source light following system is respectively in signal connection with the objective lens and the LED light source. The control module is used for controlling the LED light source to be turned on or off. According to the utility model, the problem of cell phototoxicity is solved by lighting a light source only when the light following system controls imaging, the problem of mismatching of light and an objective lens is solved through the LED array, and based on an array LED circuit principle, the functions of light adjustability, constant current, RS485 control and the like are added, so that the light brightness of the LED array is more stable, and the light intensity of the objective lens is more stable. According to the invention, the LED light source can not change along with factors such as external voltage and temperature, has a more stable light supplement effect, realizes control of stable lighting of each lamp bead at any brightness, and improves the stability of light brightness and the light supplement effect.

The invention provides a living cell imaging device capable of continuously loading physiological pressure and an assembling method and application thereof.The living cell imaging device comprises a box body and a box cover arranged on the box body, the box cover is provided with a first threaded hole and a Luer-lock gas connector, the middle of the box body is provided with a hollow part serving as an observation window, and the box cover is provided with a second threaded hole and a Luer-lock gas connector; a plurality of first threaded holes are formed in the box cover, the first threaded holes are annularly formed along the edge of the box cover, the box cover and the box body are fixed by screws through the first threaded holes, the Luer-lock gas interface is connected with an external gas supply device such as a gas cylinder through a gas pipe, and the gas pipe is connected with the gas supply device through a pressure reducing valve, a pressure gauge and the like. According to the scheme, high-resolution real-time microscopic observation of cells can be continuously performed during pressurized culture, a commercial culture dish and a microscope are compatible, the assembly process is standardized, and mechanical conduction events can be dynamically captured.

The invention relates to a method for efficiently detecting the endocytosis activity of an antibody through combined application of a nano antibody and a living cell imaging system and application, and the method comprises the following steps: incubating the nano antibody coupled with a pH-sensitive luminous group and a sample to be detected, and then detecting by adopting the living cell imaging system. On the basis of an Inucyte living cell imaging system, a high-affinity nano-antibody for recognizing an Fc region of a detected antibody is utilized, a pH-dependent fluorescent probe coupled to the nano-antibody can emit a fluorescent signal under acidic environment conditions such as lysosome and the like, and through high-flux continuous detection, the detection sensitivity is high, and the detection sensitivity is high. And detecting the endocytosis fluorescence signals of the same tumor cell sample at different time points, and evaluating the endocytosis activity of the monoclonal antibody for constructing the ADC drug.

The present application provides a system and method for live cell RNA imaging and sensing. The system of the present application comprises: (i) a target recognition module comprising a circular binding RNA, the circular binding RNA comprising at least one target binding sequence configured to form a double-stranded binding region by complementary binding with a sequence on a target RNA; and (ii) a signal module comprising a double-stranded RNA binding protein and a signal element, the double-stranded RNA binding protein being connected to the signal element, wherein the double-stranded RNA binding protein recognizes and binds to the double-stranded binding region. The method of the present application comprises introducing the system into a live cell and performing imaging and detection. The system of the present application has the advantages of non-invasiveness, low fluorescence background and universality.

1. Name of the product in this design: Live Cell Imaging Monitor. 2. Purpose of this design: For long-term dynamic monitoring and analysis of living cells. 3. The key design feature of this product is its shape. 4. The image or photograph that best illustrates the design's key points: 3D view 1.

The invention belongs to the technical field of fluorescent carbon quantum dots, and particularly relates to a double-color double-targeting carbon quantum dot as well as a preparation method and application thereof. The surface functional groups of the carbon quantum dots are rich and easy to modify, the surface functional groups can be changed through ion modification, so that single mitochondrial targeting is converted into double-color double-targeting of mitochondrial and lysosome, the cell imaging effect is good, and the purpose of observing interaction of different organelles is achieved. The double-color double-targeting carbon quantum dot provided by the invention does not need to be dyed by two fluorescent probes with good redyeing property at the same time, so that the multi-probe dyeing operation is simplified, and the influence factors on cells are reduced. The double-color double-target material provided by the invention is accurate in positioning and strong in light stability, and can keep good stability before and after ion modification. The double-color double-targeting carbon quantum dot provided by the invention has low toxicity to living cells, and is suitable for long-time living cell imaging.

The present application relates to the technical field of chemical genetics marking, in particular to a kind of probe for marking quantifying protein dynamics.The probe includes the compound of the following structural formula: the compound contains halogenated alkane chain, can be specifically covalently combined with halogenated alkane dehalogenase-HaloTag label protein.It is combined with the fusion protein containing HaloTag in cell with the purpose of protein, with high specificity and stability, low toxicity, with good biocompatibility and strong anti-photobleaching ability in live cell imaging.The compound has the nature of space limited emission effect, through HaloTag bridge, can sensitively mark and respond to the spatial dynamic change of any single protein molecule in cell physiological process, and then realize the quantification and visualization of protein dynamics in method, for recording physiological activity under different time scales.

The present application relates to the field of bioengineering, and particularly relates to a kind of artificial codon optimization nucleic acid molecule of the red fluorescent protein coding.The present application provides nucleic acid molecule of red fluorescent protein coding, and the red fluorescent protein is mCherry, compared with the existing red fluorescent protein nucleotide sequence, the number of CpG site is increased from 56 to 92, the content of the third position of codon G / C (GC3) is not less than 99%.The nucleotide sequence provided by the present application is the mCherry synonymous codon substitution sequence obtained by combining the increase of the number of CpG sites and the use of optimal codon strategy, which is 1.3-2.4 times higher than the fluorescence intensity of the sequence before modification in mammalian (human / mouse) cells;The present application uses the method of codon optimization to obtain the nucleic acid molecule of the red fluorescent protein that does not exist in nature, which can encode the red fluorescent protein with stronger fluorescence intensity, and promotes the development of live cell imaging.

The invention discloses a construction method of a fish nocardia disease pathogenic mechanism model, and belongs to the field of biotechnology and disease models. The invention aims to solve the technical problem that a deep causal mechanism is difficult to reveal due to the fact that microscopic dynamic interaction of pathogens and hosts cannot be observed in real time and in situ when an existing animal model and an in-vitro cell model are used for researching the disease. The method comprises the following steps: constructing a fish immune organ in an intelligent micro-fluidic chip integrated with a micro-actuator as a high-bionic-degree physical experiment platform; the living cell imaging system is used for collecting four-dimensional image data of the organoids in the nocardia infection process in real time. According to the method, through normal form transformation from passive observation to active intervention, the microscopic process of the disease can be reproduced in a high-fidelity manner and is regulated and controlled prospectively, so that a causal chain between key pathological events is deeply revealed, and a powerful intelligent research platform is provided for developing a new disease prevention and treatment strategy.

The present invention provides a novel one-step synthesis method for producing cyanine-based fluorophores with modified polymethine chains to prevent aggregation, enhancing their utility in biomedical research. These fluorophores, such as unsymmetrical heptamethine cyanines like SAT-IR-746, exhibit improved sensitivity, photostability, and brightness, making them ideal for applications such as live-cell imaging, biomolecule labeling, and near-infrared fluorescence microscopy. The synthesis process involves the preparation of a pyridinium benzoxazole (PyBox) intermediate from a substituted pyridine derivative and 2-chlorobenzoxazole, followed by condensation with sulfonated and substituted indolenines to form asymmetrical cyanine dyes, achieving high yields (e.g., 55-82% for SAT-IR series compounds). By addressing the aggregation issue common in traditional cyanine dyes through β-position substitution on the polymethine chain, this invention enables more accurate and reliable labeling and tracking of biomolecules without quenching, facilitating advanced imaging techniques and deeper insights into biological processes.

The application belongs to the technical field of live cell imaging, and discloses a live cell nucleic acid imaging system, method and use based on CRISPR and nanobody, which comprises: a modified dCas9 protein expression vector containing n ALFA polypeptide tags; a modified sgRNA expression vector comprising a specific guide sequence; and a fluorescent nanobody expression vector containing a bivalent or trivalent fluorescent nanobody coding sequence, which are operatively connected, wherein a fluorescent protein is inserted between nanobody split sites as an internal tag, can be combined with the ALFA polypeptide tag carried by the dCas9 protein and stably emit light, and the free state fluorescent nanobody is self-degraded and does not emit light. The application can visualize single copy nucleic acid sequences with single molecule resolution and high signal-to-noise ratio, especially the imaging marking of single copy nucleic acid sequences in live cells.

A sample monitoring structure includes fluorescently-labeled hydrogel that is size-responsive to changes in one or more properties of aqueous liquid medium contacted with the sample monitoring structure. An intensity of a fluorescent emission from the sample monitoring structure changes with changes in the size of the fluorescently-labeled hydrogel. The sample monitoring structure is advantageously used in automated processing and analytical evaluation of large numbers of fluid samples over extended periods of time analytical evaluation systems, including flow cytometry systems, live cell imaging and analysis systems and parallel bioreactor system. The sample monitoring structure may be in a form of sample monitoring beads for addition to liquid media of fluid samples subjected to analytical evaluation.

The application belongs to the technical field of live cell imaging, and particularly relates to a live cell imaging system based on CRISPR-dCas12a and fluorescent RNA aptamer and application thereof. The application discloses a CRISPR LiteColor system, which mainly comprises CRISPR-dCas12a and crRNA fused with fluorescent RNA aptamer. When dCas12a and the modified crRNA are cooperatively targeted to a target site of a genome, and a corresponding small molecule fluorescent dye is added, the visualization imaging of the target gene site can be realized. The system embeds the fluorescent RNA aptamer into the stem loop structure region of the crRNA, and only relies on a single crRNA to target and recognize the specific site of the TTTV type PAM sequence in the genome, has the characteristics of autonomous degradation and low cell transfection load, and greatly simplifies the experimental operation process on the basis of ensuring high specificity of targeted recognition.

Systems and methods for predicting an immune response against a tumor in a patient having the tumor are provided. The relative mass or changes of mass of tumor cells or immune cell in the tumor can be ex vivo observed, and an immune status of the tumor can be determined based on the mass of tumor cells or immune cell. The immune status can provide a guidance to predict the immune response against the tumor in the patient.

1. Name of the product in this design: Intelligent Live Cell Imaging Analyzer. 2. Purpose of this design: This design is used for imaging observation and real-time image acquisition of live cells. 3. The key design features of this product are its top view. 4. The image or photograph that best illustrates the design's key features: a top view.

Disclosed are a cyanine-derived compound, a preparation method therefor, and application thereof. The compound has the structure represented by formula (1), and the preparation method for the compound is further disclosed. The series of compounds of the present invention can be used as fluorescent markers for live-cell imaging analysis or flow cytometric analysis, solving problems of high toxicity, high cost and poor imaging effect of current.

The invention relates to a portable quantitative phase microscope for living cell imaging, comprising: a light source module for providing a coherent illumination beam; the interference light path module is used for dividing the illumination light beam into a sample light beam and a reference light beam and enabling the sample light beam and the reference light beam to interfere; the sample bearing module is used for bearing a to-be-tested sample; the imaging module is used for collecting the light penetrating through the sample and imaging the light; the image acquisition and processing module is used for acquiring an interference pattern and reconstructing a quantitative phase image; the light source module, the interference light path module, the imaging module and the image acquisition and processing module are integrated in a compact shell to form portable equipment. The system has the remarkable advantages of compact structure, low cost, convenience in use, high imaging precision, non-staining real-time imaging and the like, realizes real portability and practicability while keeping high imaging quality, and is suitable for various application scenes such as scientific research, teaching, portable biological detection and the like.

The application discloses a fluorescein probe molecule targeting VISTA immune checkpoints and a preparation method and application thereof. The probe can block the interaction with ligands and inhibit the signal path by high-selective combination with VISTA proteins, and realizes the continuous tracking of the VISTA positioning and dynamic change on the premise of not interfering with the physiological state of cells. The probe has the living cell imaging ability with the minute-level resolution, is compatible with the confocal microscope, various fluorescence microscopes and flow cytometry and the like technologies, is suitable for the membrane dynamic analysis and high-throughput screening, and provides a new tool for the research on the immune regulation mechanism and the drug development.