21 results about "Patch clamp" patented technology

A method and system for differentiating acute and chronic pain based on the functional state of specific neural circuits is proposed. This method utilizes in vivo two-photon / endoscopic calcium imaging and ex vivo brain slice patch-clamp techniques to longitudinally monitor the activity dynamics of neurons in the DRN and ZI brain regions during the acute and chronic phases of a neuropathic pain model. By extracting and analyzing calcium and electrophysiological signal parameter sets, distinguishing criteria are established: if the activity of 5-HTergic neurons in the DRN is specifically enhanced in the acute phase, the indication is acute pain; if the activity of GABAergic neurons in the ZI is specifically enhanced in the chronic phase, the indication is chronic pain. This invention achieves an objective and precise differentiation of the transition from acute to chronic pain at the level of specific neural circuits, providing a new strategy and tool for pain mechanism research and clinical diagnosis and treatment.

The invention relates to the technical field of gene detection devices, in particular to a silicon nitride nanopore device and a preparation method and application thereof based on an angstrom hole, a helium ion beam is used for irradiating the surface of a silicon nitride film of a cleaned silicon nitride chip to obtain a silicon nitride chip with an angstrom hole, and the silicon nitride chip is used for detecting the gene detection device. And after cleaning and drying, carrying out dielectric breakdown to obtain a nanopore with a required size, measuring the obtained nanopore again by using the patch clamp, and drying by using nitrogen to obtain the silicon nitride nanopore device. According to the silicon nitride nanopore device and the preparation method and application thereof based on the angstrom pore, the generation position of the nanopore is controlled in a helium ion beam irradiation area through an angstrom pore confinement intrinsic carrier effect and an avalanche effect and breakdown critical defect density correlation mechanism; and then the formation, evolution and size regulation and control of the nanopore are tracked in real time through dielectric breakdown ion current, and controllable manufacturing of the nanopore with the extremely small size is achieved.

This application provides a nanoprobe-based electrophysiological signal and biochemical biomarker detection system, comprising: a patch-clamp amplifier; a working electrode electrically connected to a signal acquisition module, the surface of which, except for the tip, is covered by an insulating protective layer, forming a GΩ-level impedance seal after implantation into the cell; a counter electrode connected to a BATH port, with its active end placed in the matrix environment of the target cell, forming a current loop; a reference electrode connected to a REF port, with its active end placed in the matrix environment of the target cell to provide a potential reference point; the patch-clamp amplifier is configured to operate in voltage-clamp mode, with the REF port connected to the reference electrode as a voltage sensing terminal, and the BATH port connected to the counter electrode as a current output terminal, simultaneously acquiring the electrophysiological signal of the target cell and the current signal corresponding to the concentration of the target biochemical biomarker. This application provides a powerful analytical tool for a deeper understanding of the electrochemical signal coupling mechanism of neurons.

The invention relates to the technical field of biosensors, and particularly discloses a C reactive protein composite nanopore sensor for slowing down the nucleic acid pore passing rate and a preparation method of the C reactive protein composite nanopore sensor. According to the method, a solid nanopore chip with a silicon nitride suspended film with the thickness of 30 nm is installed in a fluid pool, and nanopores with the diameter of 8-10 nm are formed in the silicon nitride suspended film; adding an electrolyte solution with the pH value of 7-9 into a Cis side chamber and a Trans side chamber of the fluid pool; two Ag / AgCl electrode wires are respectively inserted into a Cis side chamber and a Trans side chamber, and the other two ends of the electrode wires are respectively connected with patch clamp current amplifiers; positive and negative voltages of 200 mV-200 mV are applied to the Trans side of the fluid pool, and a current value is recorded as a negative tapping current; adding a C reactive protein solution with the final concentration of 10-100 nM to the Cis side of the fluid pool; meanwhile, negative voltage of-50 mV to-300 mV is applied to the Trans side, the C-reactive protein composite nanopore sensor is obtained when it is observed that long-time current is smaller than negative pore opening current, and the DNA pore passing speed reduction effect of nearly two orders of magnitude can be achieved.

The invention discloses a method for separating cochlea related cells in situ based on a patch clamp system. The method comprises the following steps: contacting an electrode with a target single cell under a microscope, and carrying out in-situ grabbing on the target single cell through negative pressure adsorption; the target single cell and surrounding cells are kept in a connection state before in-situ grabbing; before the electrode approaches the target single cell, positive pressure is continuously applied, and when the electrode is in contact with the surface of the target single cell, the positive pressure is instantly stopped and switched to negative pressure. According to the method provided by the invention, high-quality cochlea related cells can be obtained in situ for deep sequencing analysis, and the method can be used for basic research and accurate diagnosis and treatment of clinical hearing impairment, and has a good application prospect.

The application provides a content extraction method based on cell membrane capacitance change, and belongs to the field of cell-level micromanipulation technology, and comprises the following steps: S1, a change relationship between cell membrane capacitance and cell membrane surface area is established; S2, a change relationship between cell membrane surface area and volume is approximately established; S3, high-resistance sealing and membrane rupture of cells are completed, constant voltage cell content extraction operation is performed, and cell membrane capacitance values in the extraction process are obtained in real time through capacitance fitting; S4, content extraction amount is estimated; S5, actual cytoplasm extraction amount is calculated based on a rotary L-shaped electrode, the relationship model established in S2 is corrected, and finally, quantitative extraction of cell contents is realized. The application utilizes an electrical model of cell membrane capacitance, realizes real-time feedback of cell content extraction amount, realizes quantitative control of content extraction, can realize a cytoplasm extraction success rate of 62.5% in the case of a whole cell patch clamp structure, and the average cytoplasm extraction amount control error is less than 6% of the total cell volume.

The invention discloses a patch clamp-based regulation and control analysis method and a patch clamp-based regulation and control analysis system for electrical stimulation induced tunnel nanotube mediated cell electrical coupling, and relates to the technical field of cell biology. The patch clamp-based regulation and control analysis method for electrical stimulation induced tunnel nanotube mediated cell electrical coupling comprises the following steps: S1, sample preparation and drug intervention: adding at least one signal path inhibitor into a cell bath solution for intervention; s2, performing membrane potential imaging and data analysis; s3, applying electrical stimulation and coupling verification: applying electrical stimulation in a specific mode to one cell in the tunnel nanotube connection cell pair by using patch clamp equipment, and executing membrane potential imaging and data analysis; s4, measuring the traction force of the cells; s5, analyzing membrane potential change and traction force change; through accurate electrical stimulation and multi-parameter monitoring, the regulation and control mechanism of tunnel nanotube mediated cell electrical coupling is deeply analyzed, and an effective means is provided for research of intercellular signal transmission.

The utility model provides an SMT patch clamp, and relates to the technical field of SMT patch clamps. The SMT patch clamp comprises a base, a driving motor is fixed to one side of the base, a two-way threaded rod is fixed to the driving end of the driving motor, threaded blocks are in threaded connection with the outer surfaces of the two sides of the two-way threaded rod, and fixing blocks are fixed to the upper ends of the two threaded blocks; a connecting structure is arranged on the close sides of the two fixing blocks, and two extrusion frames and two pressing frames are arranged on the connecting structure. The connecting structure can drive the two extrusion frames to move reversely in the length direction of the base and can drive the two pressing frames to move in the height direction of the base at the same time. According to the SMT clamp, the driving motor drives the bidirectional threaded rod to rotate, so that the effect of conveniently and simultaneously clamping in two directions is achieved, the effect of improving the use convenience of the SMT clamp is achieved, the effect of removing dust by blowing is also achieved, and the effect of improving the use effect of the SMT clamp is achieved.

The application discloses a single-vacuole particle electrochemical signal recognition classification method and a recognition system, and the method comprises the following steps: S1, a carbon fiber ultramicroelectrode is prepared, and a voltage is applied in a membrane patch clamp system constant potential mode, and a picoampere-level current response generated by oxidation of single-vacuole content on the surface of the electrode is collected; S2, the collected original current signal is subjected to smoothing filtering, and a multi-parameter peak recognition method based on a threshold value is used to realize feature peak recognition and multi-dimensional physical / statistical feature extraction; S3, the extracted peak feature data is subjected to a preprocessing operation including missing value processing, standardization, principal component analysis dimension reduction and class balancing; and S4, at least one machine learning model is constructed and trained based on the preprocessed feature data, and is used for automatically recognizing the particle size class of the vacuole particle. The application combines the advantages of single-vacuole electrochemical high-sensitivity measurement and intelligent algorithm classification, and has the advantages of high recognition efficiency, good repeatability and wide applicability.

The invention discloses a portable plant surface potential detection device and method, the device comprises a detection electrode assembly, a portable electrochemical workstation and a terminal computer, the detection electrode assembly is electrically connected with the electrochemical workstation through a shielding conductive wire, and the electrochemical workstation is in bidirectional communication connection with the terminal computer. Plant surface potential monitoring is carried out by creatively adopting an open circuit electrochemical method, traditional high-price patch clamp equipment is replaced with a low-cost portable electrochemical workstation, and all parts of the device are in portable modular design and can be quickly assembled and disassembled, so that the device is convenient to use and popularize. And the field real-time monitoring of the surface potential of plant stalks and leaves in a field natural growth state is realized. According to the device, under set detection and stimulation parameters, personal errors are effectively reduced, detection data are accurate, stability is high, potential response rules of plants under stimulation of fire, damage and the like can be captured, and the device has the advantages of being low in cost, portable, high in adaptability and wide in application prospect.

The invention relates to an analysis method for mouse retinal ganglion cell morphological classification, which realizes single-cell-level accurate analysis and high-throughput integration: carrying out whole-cell patch post-clamp dendritic structure reconstruction on retinal ganglion cells, objectively quantifying parameters such as dendritic area, layering depth, dendritic asymmetry and the like, and analyzing the morphological classification of the mouse retinal ganglion cells. The subjective experience dependence of traditional Golgi staining and the resolution limitation of immunohistochemistry are thoroughly eliminated; a hierarchical classification rule is established through an automatic algorithm, manual interpretation deviation is eliminated, and the classification standardization degree is improved; the method breaks through the low-flux bottleneck of a genetic marker technology, does not need to depend on time-consuming gene model construction, synchronously analyzes the morphological diversity of cells through single cell sequencing and morphological reconstruction, and reveals that cells with similar dendritic layering positions can present functional differentiation due to transcriptome differences. The morphological analysis system established by the invention has single cell precision, high-throughput coverage and multi-dimensional relevance, and provides technical support for systematically analyzing the morphological development mechanism and subtype specificity of RGCs.

This invention discloses a method for efficient reconstruction of neuronal morphology based on a visual model. Addressing the problem of high noise and low contrast in patch-clamp stained neuronal images, which hinders automated reconstruction using existing methods, the method preprocesses the Z-axis tomographic images by grayscale conversion, frequency domain filtering, and multi-scale block segmentation. The SOLOv2 model is pre-trained using a wall seam dataset through transfer learning, and then fine-tuned using labeled neuronal data to identify slender branch structures. The segmentation results of each image block are concatenated to reconstruct a binary mask. The Z-axis masks are then superimposed, and isolated regions with fewer than three consecutive frames and a distance exceeding 70 pixels from the main skeleton are removed to obtain a three-dimensional spatially consistent mask. Finally, the width, height, maximum path distance of the cell body, and longest trunk length are automatically extracted using skeletonization and graph theory algorithms. On 207 mouse and macaque neurons, the correlation coefficient with manual reconstruction using ImageJ reached 0.88–0.98, reducing reconstruction time from several hours to seconds, achieving efficient and accurate automated reconstruction.

The application discloses arylpiperazine salt compounds, a synthesis method thereof and application thereof in antidepressants, and belongs to the field of medicinal chemistry. The arylpiperazine salt compounds containing bioactivity are obtained through a plurality of continuous reactions such as N-alkylation, protection of a group, condensation and deprotection, etc. of arylpiperazine as raw material. Researches show that the compounds exhibit high receptor binding of 5-HT 2A R and GABA A Rs. The target points are confirmed through molecular docking, micro-thermal mobility instrument, surface plasmon resonance, inositol-phosphoric acid (IP1) accumulation and cell membrane patch clamp, etc.; and the anti-inflammatory activity of the compounds is evaluated by enzyme-linked immunosorbent assay, so as to provide a potential active structure skeleton for development of antidepressants.

The present application relates to the application of isoquercitrin in the preparation of a drug for treating post-traumatic stress disorder, and belongs to the technical field of natural chemical medicine development. The present application systematically clarifies the whole-chain mechanism of isoquercitrin in treating PTSD through the "GABA A R-PINK1 / Parkin-mitophagy" pathway by means of a variety of technical means such as molecular docking, microthermal swimming, patch clamp, proteomics, cell and animal models. Compared with the clinically commonly used drugs for treating post-traumatic stress disorder, isoquercitrin, as a natural flavonoid compound, can improve the symptoms of PTSD while not damaging the learning and memory function, and has better safety. Moreover, isoquercitrin is widely sourced and has a mature preparation process. The present application provides a theoretical basis and experimental support for the application of isoquercitrin in the development of drugs for treating post-traumatic stress disorder, and has important clinical application value and industrialization prospects.

This invention discloses a method for detecting viral or pseudoviral cell infection, comprising: applying a sinusoidal voltage to an attached membrane in patch-clamp mode and detecting the corresponding membrane current; inducing viral or pseudoviral cell infection by binding of viruses or pseudoviruses in the electrode fluid to receptors on the attached membrane; performing phase discrimination demodulation on the corresponding membrane current signal using the phase of the input sinusoidal voltage as a standard to obtain the imaginary and real parts of the membrane current and calibrating and standardizing them in real time; calculating the membrane capacitance of the attached membrane based on the real and imaginary parts of the current signal, based on the calibration and standardization; identifying viral or pseudoviral cell infection events and their modes based on the dynamic characteristics and polarity of the membrane capacitance changes; and estimating the size of the virus or pseudovirus based on the magnitude of the membrane capacitance changes. This invention enables real-time, high-precision detection of various viruses or pseudoviruses infecting host cells via envelope-cell membrane fusion or endocytosis in acutely isolated cells, cultured cells, and isolated tissue cells.

The present disclosure relates to methods, apparatuses, and systems to measure one or more electrical signals from a cultured cell layer. In some embodiments, such electrical signals can be indicative of transport or lack of transport of an agent through the cell layer.

The invention belongs to the technical field of neurobiology, and particularly relates to a targeted NPTX1-Kv7.2 signal axis for inhibiting neuronal hyperexcitation and improving memory impairment formed by aging. According to the invention, a scene fear memory mouse behavior model is adopted, experimental research is carried out by combining in-vitro brain slice patch clamp recording, single-molecule fluorescence in situ hybridization, immunohistochemistry and other methods, and the result shows that NPTX1 can be used as a cognitive disorder molecular marker, and neuronal hyperactivity caused by aging can be rescued by specifically supplementing NPTX1 in a DG brain region; the activation of DG neurons during scene fear experience is improved, and neuronal activation damage and dysmnesia caused by aging are improved; the retigabine serving as an agonist of the NPTX1 interaction protein Kv7.2 is injected in the intraperitoneal cavity, so that neuron activation damage and dysmnesia caused by senescence can also be effectively improved. The invention discloses the effect of the NPTX1-Kv7.2 signal axis in improving the memory impairment of an aging individual, and provides a new thought and target spot for treating the memory impairment caused by aging.

The invention discloses application of orexin-A in preparation of a medicine for promoting motor function recovery after spinal cord injury. It is found for the first time that orexin-A has a promoting effect on exercise recovery after spinal cord injury, which may involve a GABA-A receptor instead of a glycine receptor. Secondly, patch clamp experiments find that the flip potentials of the two currents in the separated neurons after SCI are significantly reduced. In addition, spinal cord injury has significant influence on the kinetic indexes of gamma-aminobutyric acid (GABA) and glycine current, and only the change of the GABA current caused by spinal cord injury is significantly reversed after orexin-A is given. In a word, the orexin-A combined GABA-A type receptor may participate in motor function recovery after spinal cord injury, but does not relate to a glycine receptor. Orexin-A may promote motor function recovery by eliminating the influence of spinal cord injury on spinal cord abdominal horn neuron GABA current characteristic indexes.

This invention proposes a plating method to improve the stability of hERG-HEK293 cells during patch-clamp assays, comprising the following steps: S1, placing cell slides in a culture dish, coating them with matrix gel for 10-30 min, and then aspirating the coating solution; S2, adding hERG-HEK293 cell suspension to the matrix gel-coated cell slides prepared in step S1, incubating in an incubator for 5-20 min, and then adding culture medium to cover the slides for cell culture; S3, removing the cell slides from the culture dish and placing them in a culture dish containing extracellular fluid, then removing the slides and placing them in a chamber filled with extracellular fluid for patch-clamp assays. This invention uses matrix gel to coat the cell slides, increasing cell stability, ensuring strong cell adhesion, reducing the risk of cell washout during perfusion, improving the cell perforation rate, and extending the cell recording time.

The invention provides an electrophysiological patch clamp system comprising a magnetic mechanical force device, and relates to the technical field of bioelectrophysiological detection, the system comprises an electrophysiological patch clamp objective table, the magnetic mechanical force device, a signal acquisition and processing module and a control module; the electrophysiological patch clamp objective table is used for placing a magnetic induction cell sample to be detected; the magnetic mechanical force device is arranged on the electrophysiological patch clamp objective table and is used for generating a static magnetic field and a dynamic magnetic field; the signal acquisition and processing module is connected with the electrophysiological patch clamp and is used for acquiring an original current signal acquired by the electrophysiological patch clamp while the magnetic mechanical force device is started and processing the original current signal so as to determine the magnetic induction characteristic and the electrophysiological mechanism of the magnetic induction cell sample; the control module is used for controlling the magnetic mechanical force device so as to adjust the magnetic field generated by the magnetic mechanical force device, and the reliability and accuracy of the electrophysiological patch clamp in a magnetoelectric experiment are improved.