77 results about "Live cell imaging" patented technology

This disclosure provides a novel method and system for characterizing morphodynamic profiles of objects, such as biological entities. This disclosure provides a shape, appearance, and motion (SAM) phenotype Observation Tool (SPOT). SPOT establishes a standardized SAM “phenome,” image descriptors resembling single-cell transcriptomes, to comprehensively quantify a cell's instantaneous state without prior knowledge. SPOT also establishes a standardized workflow for temporal analysis. SPOT is a generalist tool, applicable to any live-cell imaging and advances biomedical discovery through its standardized, unbiased, streamlined workflow to quantify phenotypic heterogeneity and predict phenotype-genotype-function coupling.

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

The invention discloses a vesicle transportation path prediction system based on artificial intelligence, and relates to the technical field of vesicle transportation, which comprises the following steps of: accurately extracting a complete time sequence movement track of a single vesicle before and after disturbance by virtue of fluorescence living cell imaging under a disturbance experiment condition in combination with a target detection model YOLOv8 and a depth correlation tracking algorithm DeepSORT; the problem that perturbation conditions are insufficiently perceived is solved, through a multi-modal data matrix, the model has the time sequence perceiving capacity for changes of a local space structure where vesicles are located, so that the sensitivity and modeling capacity of the model for track variation in a complex environment are remarkably improved, deep fusion of multi-modal features is achieved by building a double-branch fusion model, and the accuracy of the model is improved. The problem that an existing prediction model cannot actively recognize and correct prediction deviation is effectively solved, and the stability and the actual usability of the system are improved.

The invention relates to the technical field of cytobiology and biomedicine detection, and discloses a time sequence fluorescence tracing system based on a coupling detection lipid probe. Comprising an imaging detection module which is used for adding FM lipophilic styrene fluorescent dye in a culture environment, forming a membrane probe to observe the morphological change of a membrane and detect the formation of vesicles, and completing the complete time sequence tracing of the cell endocytosis process by adopting a content dyeing method; the deep learning module is used for carrying out deep learning on the obtained time sequence image, establishing a living cell imaging screening and image analysis system, and identifying protein molecules related to the target external vesicles; carrying out image description on the generation, transportation and fusion processes of the vesicles generated by the proteins in different stages before fusion of the outer vesicles and the cell membranes and after fusion and shearing; and the knock-down operation module is used for exploring the generation mode of the vesicle contents in the early endosome in combination with knockout and knock-down operations of the specific drug compound.

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 invention belongs to the technical field of cell culture meat screening, and particularly relates to a cell culture meat food hazard factor screening system and device.The cell culture meat food hazard factor screening system comprises a sensing unit, an execution unit, an analysis unit and a control unit; the sensing unit comprises an integrated sensor and living cell imaging equipment, the integrated sensor is used for collecting the culture environment of cell culture meat in real time, and the living cell imaging equipment is used for dynamically tracking the cell growth state; the execution unit is used for executing sample data acquisition and detection according to presetting; and the analysis unit analyzes and processes the data information sent by the sensing unit and the execution unit based on an AI data analysis platform. The intelligent level of screening the harmful factors of the cell culture meat can be greatly improved, so that dynamic evaluation and early warning of the harmful factors in the production process of the cell culture meat are realized, workers can timely and effectively perform corresponding treatment, and the production quality of the cell culture meat is further ensured.

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 belongs to the technical field of RNA detection, and particularly relates to an alkenyl indole derivative, a preparation method thereof, and an application in the field of RNA fluorescent probes. The alkenyl indole derivative uses alkenyl indole as the mother nucleus and is a novel functional fluorescent probe, which can stain RNA in cells and broadens the range of types of fluorescent probes. The process of using this compound as a fluorescent probe to detect RNA is not interfered by other components, showing excellent anti-interference performance and high fluorescence intensity. In addition, its preparation process is simple, the raw materials are easily available, the reaction conditions are mild, the product structure is stable, and it is convenient for storage. It has important application value in the fields of RNA detection and its biological function research, and particularly shows broad application prospects in the fields of live cell imaging, molecular diagnosis, etc.

The invention discloses a dicyanomethylene-4H-pyran-based fluorescent probe DCM-Pro as well as a preparation method and application thereof, the probe realizes fluorescence intensity and fluorescence lifetime response to polarity change of a microenvironment by optimizing a molecular structure, and can be covalently bound with protein through a Michael addition reaction. The polarity sensitivity of the probe is adjusted by regulating and controlling the proportion of local excited states in the intramolecular charge transfer process. The fluorescence lifetime of the DCM-Pro is linearly related to the polarity of the microenvironment, and the change of the polarity of the microenvironment in the protein aggregation process can be quantitatively detected. The probe is a fluorescence lifetime probe sensitive to polarity and can be used for visual detection of iron-mediated protein aggregation. The probe has good biocompatibility and specificity, is suitable for living cell imaging, provides a new tool for researching protein aggregation in the ferroptosis process, and also provides a new method and thought for mechanism research and drug screening of neurodegenerative diseases.

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.

The utility model discloses a high-throughput living cell imaging analysis system, which comprises a bottom plate and a sample table above the bottom plate, a first platform is slidably arranged at the top of the bottom plate along the X direction, the first platform is driven to slide by a first lead screw motor, a second platform is slidably arranged on the first platform along the Y direction, and the second platform is driven to slide by a second lead screw motor. A vertical sliding block is arranged at the top of the second platform in a sliding mode in the Z direction, the vertical sliding block is driven by a precise lead screw motor to slide, and a fluorescence imaging mechanism is arranged on the vertical sliding block. The whole set of instrument can be placed in a CO2 culture box, so that long-time living cell imaging of cells can be realized in a proper culture state. Different experiment conditions can be independently set for each cell culture container, and various experiments can be simultaneously realized in the same environment.

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 utility model discloses an electric adjusting carrier applied to a living cell imager, which comprises a frame main body, an objective table arranged on the frame main body and a perforated plate outer frame arranged on the objective table, and a group of driving motors and an XY axial linkage mechanism are arranged in the frame main body. The group of driving motors drive the XY axial linkage mechanism to work, and the perforated plate outer frame is connected with the XY axial linkage mechanism. According to the utility model, the XY axial linkage mechanism is concentrated in the frame main body, and the XY axial linkage mechanism is controlled by a group of motors, so that the precise alignment of the perforated plate outer frame in the XY axial direction can be realized, and the alignment precision and the convenience degree of alignment control can be improved; due to the highly concentrated design, the carrier is small in size, the overall size is not influenced when the carrier is carried on the living cell imager, the working distance of the imager in the Z-axis direction is not influenced, and the carrier is attractive and elegant; the XY axial linkage mechanism is mainly composed of a guide rail, a pull rope, a sliding block and other conventional parts, and the production cost and difficulty can be reduced.

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.

A method for observing biomolecular compensates by providing a cell expressing a first fusion protein comprising a protein that forms a component of the biomolecular condensate and a second fusion protein that comprises a nuclear localization signal is provided. The present invention further provides for methods and means for screening candidate agents by exposing cells expressing a first and second fusion protein with said candidate agents and observing the effects on biomolecular condensates.

The invention provides a fluorescent compound and application thereof in preparation of a specific imaging biological sample autophagy process probe or an autophagy regulator screening preparation, and belongs to the technical field of biological medicine. The fluorescent compound provided by the invention comprises a fluorophore, a linking group and an autophagy targeting group which are covalently bonded in sequence. The fluorescent compound provided by the invention is a small-molecule chemical probe, and is combined with an autophagy marker protein, namely a microtubule-associated protein light chain 3 (LC3 protein) in an affinity manner to realize high-specificity detection of an autophagy process; a dynamic signal dependent on the autophagy process is presented, fluorescence is turned on after combination with LC3 protein, and fluorescence changes color after fusion of an autophagosome and a lysosome, so that high-specificity, high-contrast and high-sensitivity living cell imaging detection of the autophagy process can be realized.

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 present invention belongs to the field of biomolecular technology, and in particular to a kind of RNA-modified live cell imaging system and its composition and application. The RNA-modified live cell imaging system comprises an RNA reporter system and a FRET probe, wherein the RNA reporter system comprises a tetracycline-induced expression system, an mMaroon1 fluorescent protein, an NLS-nuclear localization sequence and a 22-repeat m6A-MS2 reporter sequence; the FRET imaging probe comprises a YTH domain, a YPet fluorescent protein, a long-chain flexible linker, an ECFP fluorescent protein, an MCP protein and an NLS nuclear localization signal. The m6A-modified live cell imaging system developed by this patent can perform high-resolution real-time dynamic imaging of m6A modification in living cells, and can realize triple dynamic monitoring of m6A modification levels and RNA localization and translation, thereby building a research platform and drug screening platform for m6A modification pathways.

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 discloses a water-phase high-brightness luminous cluster-DNA compound and a preparation method and application thereof, and the preparation method comprises the following steps: A1, carrying out mixed incubation on single-stranded DNA with different lengths and different base sequences and hydrophobic metal nanoclusters, analyzing the product through agarose gel electrophoresis, and screening out the optimal single-stranded DNA; a2, hydrophobic metal nanoclusters and the single-stranded DNA screened in the step A1 are mixed according to a certain proportion, the hydrophobic metal nanoclusters are mediated by the single-stranded DNA to be transferred from an organic phase to a water solution phase, and the cluster-DNA compound which is good in water phase dispersity and high in brightness light emitting can be prepared. The luminous cluster-DNA compound prepared according to the invention has the advantages of good water dispersibility, small and controllable size, high luminous efficiency, strong light stability and good biocompatibility, has significant advantages in living cell imaging, and provides a new tool for biological imaging.

The invention relates to the field of bioengineering, in particular to an artificial codon optimized nucleic acid molecule for coding red fluorescent protein. The invention provides a nucleic acid molecule for coding red fluorescent protein, the red fluorescent protein is mCherry, compared with the nucleotide sequence of the existing red fluorescent protein, the number of CpG sites of the nucleic acid molecule is increased from 56 to 92, and the content of G / C (GC3) at the third site of a codon is not less than 99%. The nucleotide sequence provided by the invention is an mCherry synonymous codon substitution sequence obtained by combining increasing the number of CpG sites and using an optimal codon strategy, and the fluorescence intensity of the synonymous codon substitution sequence in mammalian (human / mouse) cells is improved by 1.3-2.4 times compared with that of a sequence before modification; nucleic acid molecules which do not exist in the nature and can encode red fluorescent protein with higher fluorescence intensity are obtained by using a codon optimization method, and the development of living cell imaging is promoted.

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 cell-stretching device which is suitable for performing in vitro stretching experiments by means of automated high-resolution live cell imaging on living cells or the tissues thereof, by applying the cells to be examined onto the cell culture surface of an elastic membrane. The invention also relates to a method for performing in vitro cell-stretching experiments by means of such a cell-stretching device in a multimodal manner (i.e. in various stretching modes including radial / equibiaxial, uniaxial, biaxial, and in different orientations of the main stretching directions).