138 results about "Single-cell analysis" patented technology

Methods are provided for diagnosis and prognosis of disease by analyzing expression of a set of genes obtained from single cell analysis. Classification allows optimization of treatment, and determination of whether on whether to proceed with a specific therapy, and how to optimize dose, choice of treatment, and the like. Single cell analysis also provides for the identification and development of therapies which target mutations and/or pathways in disease-state cells.

The invention provides a method of making measurements on individual cells of a population by forming reactors containing single cells and a predetermined number, usually one, homogeneous sequence tag. In one aspect, the invention provides a method of making multiparameter measurements on individual cells of such a population by carrying out a polymerase cycling assembly (PCA) reaction to link their identifying nucleic acid sequences, such as sequence tag copies derived from a homogeneous sequence tag, to other cellular nucleic acids of interest, thereby forming fusion products. The fusion products of such PCA reactions are then sequenced and tabulated to generate multiparameter data for cells of the population.

The invention relates to a micro-fluidic chip and a micro-fluidic chip system for single cell analysis and a single cell analyzing method. The micro-fluidic chip for single cell analysis comprises a transparent top cover, a microfluid channel, a substrate with a micropore array and a bottom plate with a cavity body, which are sequentially connected from top to bottom in a sealing manner, wherein the transparent top cover is provided with an inflow hole and an outflow hole which are used for communicating the outside and the microfluid channel, the micropore array of the substrate is arranged below the microfluid channel and is communicated with the microfluid channel, each micropore is internally provided with an electrode, each electrode is provided with a cathode and an anode, each micropore can contain a single cell only, the bottom of each micropore is provided with a channel which is communicated with the cavity body of the bottom plate, and the cross section of the channel is smaller than the size of each single cell. With the adoption of the micro-fluidic chip, the micro-fluidic chip system and the single cell analyzing method, the capturing, identifying and disruption processes of each single cell are integrated on one micro-fluidic chip; as a corresponding operation method is provided, the content of each single cell is rapidly acquired and accurately analyzed.

Disclosed herein are methods for single-cell sequencing. In some examples, the methods include enriching a sample comprising a plurality of cells for cells of interest to produce an enriched cell sample; isolating one or more cells of interest in the enriched cell sample; and obtaining sequence information of one or more polynucleotides from each of the one or more isolated cells. Obtaining sequence information may include generating a molecularly indexed polynucleotide library from the one or more isolated cells. Enriching the sample may include focusing cells of interest in the sample using acoustic focusing.

The invention discloses a method and an automatic instrument device for enriching and extracting target cells and separating single cells by using a positive magnetic bead method and performing immunity and molecular biology identification and analysis on single cells. By the method and the instrument device, the on/off of a capture magnet and a release magnet is controlled by various methods to complete target cell searching, capturing, cleaning and releasing operation once or for multiple times, so that the target cell detection sensitivity and stability are improved. When the capture magnet searches and captures the target cells, the search line is circular, square, comb-shaped, S-shaped or U-shaped. In addition, the captured substances are filtered, most free micro magnetic beads are removed, the purity of the product is further improved and the product can be used as a good experimental material. By combining a special filter and the adsorption of the capture magnet, more than 95 percent of free micro magnetic beads can be effectively removed. Meanwhile, the types of the single cells are identified and biological characteristics of the single cells are analyzed by an immunofluorescence staining method and a method in molecular biology, and effective biological indexes are provided for clinical diagnosis and treatment of cancers.

Described here is an automated robotic device that isolates circulating tumor cells (CTCs) or other biological structures with extremely high purity. The device uses powerful magnetic rods covered in removable plastic sleeves. These rods sweep through blood samples, capturing, e.g., cancer cells labeled with antibodies linked to magnetically responsive particles such as superparamagnetic beads. Upon completion of the capturing protocol, the magnetic rods undergo several rounds of washing, thereby removing all contaminating blood cells. The captured target cells are released into a final capture solution by removing the magnetic rods from the sleeves. Additionally, cells captured by this device show no reduced viability when cultured after capture. Cells are captured in a state suitable for genetic analysis. Also disclosed are methods for single cell analysis. Being robotic allows the device to be operated with high throughput.

The invention discloses a full-automatic single particle/single cell capturing chip based on a digital microfluids technique and application thereof. The full-automatic single particle/single cell capturing chip comprises a digital microfluids chip with multiple pattern layers and an integrated circuit, wherein the digital microfluids chip with multiple pattern layers consists of an upper electrode plate and a lower electrode plate; the upper electrode plate adopts lyophobic conductive glass as a ground electrode; the integrated circuit is used as a digital microfluids circuit control system.The full-automatic single particle/single cell capturing chip has the advantages that the single particles/single cells can be fully automatically captured, the operation is simple and quick, and thecapturing efficiency is high; a sample can be recycled without damage, and the capturing chip is especially suitable for rare samples; the continuous and complicated parallel operation of the capturing unit is realized by program control, and the crossing pollution is avoided; the capturing chip can be widely applied to the fields of detection of single particles and analysis of single cells, suchas isolation, coupling and detection of single particles, and isolation, culture and nucleic acid amplification of single cells.

The invention discloses a two-laser induced fluorescence multi-color detector used for a single-cell electrophoretic chip.The detector is composed of a laser excitation module, a fluorescence collection and detection module, a microimaging module, a signal collection and processing module and a microfluidic chip detection platform and has the functions of performing two-laser induced fluorescence multi-color detection, normal microscopic observation, two-laser spot focusing and detection area alignment observation and the like on a microfluidic chip.By means of the detector, simultaneous detection and quantification of fluorescence substances with different light absorption and fluorescence characteristics, especially fluorescence labeling multi-component activated small molecules, with different excitation and different emission, in single cells of electrophoretic separation of the microfluidic chip can be achieved, and space of relative research and application of laser induced fluorescence detection, the microfluidic chip, single-cell analysis, detection of activated small molecules and the like is greatly expanded.

The invention relates to a micro-fluidic chip for capturing single cells/single granules through high-flux pairing and application of the micro-fluidic chip. The micro-fluidic chip comprises three parts, namely a capturing layer, a control layer and a carrier from top to bottom, wherein the capturing layer consists of two runners in parallel and a connection channel; the control layer is an isolation pump layer; a plurality of single granule/single cell capturing units are arranged inside the runners and are used for capturing single granules/single cells; the connection channel is used for communicating two paired capturing units; isolation channels are positioned below the connection channel, are perpendicular to the connection channel and are isolated by diaphragms. The micro-fluidic chip is capable of capturing single granules/single cells in a high flux manner, is high in capturing efficiency, capable of achieving pairing of single granules/single granules, single cells/single cells and single granules/single cells, high in pairing efficiency, capable of achieving parallel operation and control on paired units, such as isolation, culture, cracking and nucleic acid amplification of single cells, can be widely applied to the field of single cell analysis, and is applicable to cell samples of small numbers.

The invention discloses a capillary needle. The capillary needle comprises a tube body, the two ends of the tube body are respectively provided with an inlet and an outlet for electro-spray, a part of the tube body is bent to form a sampling end, and the sampling end is provided with a sampling port communicating with a cavity in the tube body. The invention also discloses an electro-spray ionization mass spectrometry analytical apparatus applying the capillary needle. The invention also simultaneously discloses a method applying the electro-spray ionization mass spectrometry analytical apparatus for analytical detection. The analytical apparatus is high in integration, compact in structure, easy to control, small in size and low in cost; the sample consumption is greatly reduced, and the experiment cost is decreased; an integrated sample introduction method is employed, and an sample introduction end does not have to be repeatedly placed in a sample and blank solution, such that cross contamination is reduced, and more importantly, the analytical flux is substantially improved; and through combination with an automatic sample changing system, different samples can be automatically introduced, and therefore, the capillary needle, the apparatus and the method are quite suitable for such fields as high-flux screening, unicell analysis, mass spectrum imaging analysis and the like.

An embodiment of the present invention is useful for identifying tumor cells in bladder cancer washes, pleural effusions, biliary tumor cells and circulating tumor cells in whole blood. Subsequent analysis may identify therapeutic treatments based on a single cell analysis of activatable elements in cell signaling pathways. This analysis can be useful for diagnosis, prognosis, therapy selection and monitoring of solid tumor diseases.

Interrogation of surface receptors of individual cells within a population of cell types in a single mixture is described. Each cell comprises, bound to target surface receptors on the cell, a collection of binding compositions, each comprising a distinct molecular tag. A continuous flow of cell medium containing the cells is pneumatically transported through a channel in a separation device, toward a cleavage/release zone, where specific tags correlating to each binding composition, and thus to each corresponding cell surface receptor, are released from the binding compositions on the cell surface. The method is effective to produce a separately detectable collection of tag signals for each cell.

A method for detect bacterial drug resistance by single cell analysis comprises mixing a certain amount of bacterial sample and a certain amount of antibiotic diluent on a slide, covering that slide with a cover glass and sealing with a sealant on four side to form a plate-type microbiochemical reaction system in which the bacterial and antibiotic diluent react biochemically. Bacteria were observed and located by phase contrast microscope and Raman spectra were obtained by laser scanning. Bacterial drug resistance could be obtained by analysis. The invention also provides a device for realizing the method, and the device comprising a plate-type micro-biochemical reaction system, a phase contrast microscope, a high-power optical microscope, a Raman spectrum excitation system and a Raman spectrum collection system. Bacteria were observed by phase contrast microscope and high power optical microscope, and then Raman spectra were obtained by Raman spectrum excitation system and Raman spectrum collection system to detect bacterial drug resistance.

Methods and oligonucleotide reagents for analyzing individual T cells are disclosed. In particular, the present disclosure provides methods for analyzing individual T cells using high-throughput multiplex amplification and deep sequencing of nucleic acids encoding T cell receptors (TCRs) and various other T cell phenotypic markers. The present disclosure further provides methods of reconstituting TCRs from individual T cells for functional studies, ligand discovery, or screening therapeutics.

Described herein are devices and methods for extracting cellular material from living cells and then depositing them into to a receptacle in a nanoliter scale. Using a nanopipette integrated into a scanning ion conductance microscope (SICM), extraction of mitochondrial DNA from human BJ fibroblasts and Green Fluorescent Protein (GFP) transcripts from HeLa/GFP cells was achieved with minimal disruption to the cellular milieu and without chemical treatment prior to obtaining the isolated sample. Success of the extraction was confirmed by fluorescence microscopy and PCR analysis of the extracted material. The method and apparatus may be applied to many different cell types and intracellular targets, allowing not only single cell analysis, but single subcellular compartment analysis of materials extracted in their native state.

The invention discloses a liquid drop capturing chip. The liquid drop capturing chip comprises a plurality of fluid micro channels arranged in parallel, wherein a plurality of liquid drop capturing structures are formed along the fluid micro channels at intervals; the liquid drop capturing structures comprise liquid drop entering ends and inhibition liquid drop flowing-out ends; and the inhibition liquid drop flowing-out ends are communicated with the adjacent fluid micro channels. The invention further discloses a fully integrated digital PCR chip based on a seaweed gel liquid drop and application thereof. The digital PCR chip based on the seaweed gel liquid drop, disclosed by the invention, can finish the full process from the generation of liquid drops to capture of the liquid drops and to the detection of the liquid drops. And moreover, seaweed gels are introduced; after the seaweed gels are solidified, the seaweed gel liquid drops are stable, cannot be fused and cannot affect the PCR amplification efficiency; and the method can be applied to the detection of circulating tumor DNA in the blood and other related biology and medical sciences; and due to the introduction of the seaweed gels, the application can be further expanded to single cell analysis and so on.

The invention discloses a single-cell analysis and authentication method at high flux by combining a Raman spectrum with artificial intelligence, and belongs to the technical field of cell analysis and authentication. The technical scheme of the method comprises the following steps that: firstly, obtaining living body cytosol to be detected, and preparing the cytosol into a single-cell array on achip by a microfluidic technology; then, obtaining the Raman spectrum of the single-cell array, and preprocessing Raman spectrum data; utilizing the preprocessed data to carry out model training, verification and checkout; and finally, obtaining an optimal model, and utilizing the optimal model to carry out authentication prediction on the preprocessed Raman spectrum data. The method has the beneficial effects that the microfluidic technology is used for separating single cells at high flux, the Raman spectrum of the single cell is quickly collected, an artificial intelligence technology is applied to analyze single-cell Raman spectrum characteristics, the single-cell Raman spectrum characteristics are classified and authenticated, authentication accuracy is high, sensitivity is high, andthe problems that the single cell can not be nondestructively and quickly analyzed and authenticated at the high flux at present are solved.

The invention relates to a method for single cell analysis of relative telomere length using multiplex pre-PCR followed by a qPCR (SCT-pqPCR).

The detection device includes slide glass with transparent film attached on surface, reactor, separation capillary (SC), reaction capillary with detection window (DW), luminescence detection part located at DW, and micromanipulator. Being positioned between the slide glass and the reactor, the micromanipulator is in use for grasping SC. Cell suspension pool and dissolve film pool are setup on the transparent film. Being setup on the slide glass, buffer solution pool is connected to high-voltage end. The reactor includes reaction continuous groove (RCG), and inlet connected to RCG for adding chemiluminescence reagent. RCG is connected to ground. RCG is connected to the reaction capillary. SC is setup in RCG. One end of SC is on the transparent film, and the other end is positioned at DW. Features are: easy of preparation, and operation, small size, low cost, and high detection sensitivity. The detection device is suitable for simple cell analysis, and research in area of life sciences.

The invention relates to the technical field of detection of rare cells, in particular to a microfluid control system for separating, focusing and sorting of rare cells and application. A microfluid control chip device comprising a separating unit, a focusing unit, a sorting unit and a flow resistance matching unit serves as the core of the system. The system is characterized in that the system can integrate operation like separation, focusing, single-cell analysis and sorting on the rare cells in body fluid such as blood in a continuous mode on one chip for completion, and therefore, the system can be widely applied to separation and sorting of the rare cells in the body fluid, and disease diagnosis, drug screening, individual-based treatment and other fields based on the rare cells.

A novel single-cell-level phenotypic assay is described, which can simultaneously analyze HIV-1 drug susceptibility and intrinsic replication capacity. This allows quantitative dissection of the functions of antiretroviral drugs into suppression of viral replication and selection of resistant viruses with diminished replication capacities. The disclosed assay provides a tool for the rational evaluation of treatment decisions for patients failing antiretroviral therapy and is expected to be an important part in clinical management of HIV.