54 results about "Cell trapping" patented technology

The present invention relates to a microfluidic device (1) for forming a cell aggregate comprising at least one first cell (C1) and one second cell (C2), and for individually processing selected cells of the cell aggregate: - Microfluidic channel (10); - At least one main inlet (11) for fluids containing a first cell, a second cell, and a third cell, respectively, located in the first portion (101) of the microfluidic channel; - An outlet (12) located in the second portion (102) of the microfluidic channel for controlling the flow rate of fluid within the microfluidic channel; - A first auxiliary inlet (131) for at least one first auxiliary fluid, located in the first portion (101) of the microfluidic channel upstream or downstream of the main inlet (11); - At least one cell trapping section (14) positioned between the first and second parts within the microfluidic channel; - At least one first valve to control the flow rate of the first auxiliary fluid, causing the fluid containing the first cells and the second cells to flow at a predetermined height within the microfluidic channel, thereby guiding the first cells and the second cells to the first and second capture units, respectively. Equipped with, The microfluidic device relates to a microfluidic device in which each cell capture section (14) comprises at least one first capture section (141) and one second capture section (142), each first and second capture section being sized to accommodate a first or second cell, the first and second capture sections being adjacent to each other in a direction perpendicular to the bottom (100) of the microfluidic channel, and forming a cell aggregate containing the captured first and second cells, each cell being at a different height relative to the bottom of the microfluidic channel.

The invention relates to a micro-fluidic chip for chemiluminescence imaging detection of single-cell secretions and a use method of the micro-fluidic chip. The micro-fluidic chip is composed of a single cell analysis layer and a glass substrate layer. The single cell analysis layer is prepared from polydimethylsiloxane and is provided with a capture channel, a sample introduction channel and a lantern-shaped microstructure array, the lantern-shaped microstructure comprises a small-size cell capture trap and a large-size chemiluminescence detection chamber, the small-size cell capture trap is connected with the capture channel, and the large-size chemiluminescence detection chamber is connected with the sample introduction channel. After a mixed solution of cells and a detection reagent is introduced into the capture channel, single cell capture is realized on the capture trap due to flow resistance difference; then, introducing air into the capture channel and the sample introduction channel to form a gas-in-liquid microcavity, and carrying out single cell isolation and incubation reaction; finally, the signal switch molecules and the substrate liquid are sequentially injected into the chemiluminescence detection chamber through the sample injection channel, and quantitative detection is conducted on the single-cell secretions according to chemiluminescence imaging signals.

The invention relates to the technical field of cell manipulation and analysis, and relates to an automatic single cell continuous capturing and screening system and a use method thereof. Comprising a bias liquid storage module, a cell injection module, a micro-fluidic chip, a cell capturing module, a cell screening module and an imaging system, the bias liquid storage module, the cell injection module, the cell screening module and the cell capturing module are connected with the micro-fluidic chip; the bias liquid storage module is used for generating bias flow; the cell injection module is used for providing cells; the micro-fluidic chip is used for capturing and screening cells; the cell capturing module is used for capturing cells; the cell screening module is used for screening cells; and the imaging system is used for acquiring captured cell images and performing visual analysis. Micro-fluidic pipeline double-bias-flow stable cell flow sample introduction is designed, a high-speed electromagnetic valve is matched to realize rapid control of a gas path on cell capturing and screening channel fluid, and a high-speed camera is combined to analyze cell positions in different areas in real time, so that automatic single cell continuous capturing and screening are realized.

This invention provides a probabilistic modeling method and system for characterizing cell capture performance in single-cell immunoblotting (scWB). First, considering the random cell settling process in a polyacrylamide gel microwell array under gravity, a probabilistic distribution model of the number of non-empty microwells is constructed. The evolution of the number of non-empty microwells under different cell loading amounts is analyzed, and the model's calculation results are validated using single-cell capture experimental results. Considering the constraint that trace protein bands in adjacent microwells may overlap, a probabilistic distribution model of the effective number of microwells is established, and a method for calculating the average number of effective microwells is given, revealing the variation of the average number of effective microwells with the spacing between microwell columns under different cell loading amounts. This invention provides a theoretical basis for further optimization of microwell array structures and significantly improves the cell sample utilization rate of scWB technology.

The application relates to a communication device and a cell capture dyeing instrument thereof, which comprises a frame and a placing seat for placing a to-be-communicated member, the placing seat is slidably connected to the frame, the frame is provided with a first driving mechanism capable of driving the placing seat to slide out of or into the frame; the frame is movably provided with a moving plate, the moving plate is provided with a sample outlet butt joint, and the frame is provided with a second driving mechanism capable of driving the moving plate to drive the sample outlet butt joint to be butt-jointed and communicated with a sample inlet of the to-be-communicated member in the frame. The application has the effect of improving butt-joint communication efficiency of the to-be-communicated member.

This invention provides a method and apparatus for splitting single-cell mixed sample sequencing data, relating to the field of biotechnology. The splitting method includes: capturing and sequencing single-cell suspensions using a single-cell platform; then performing reference genome alignment, cell identification, and gene expression level quantification on the sequencing data using Cellranger; splitting the cell data identified in step a into two groups of cell data for different sexes based on SNP locus information from the 1000 Genomes Project; and distinguishing the two groups of cell data from male or female samples based on the proportion of sex-specific genes expressed in the two groups of cell data. This splitting method eliminates the need for additional experimental operations such as protein labeling and genome sequencing, and can provide accurate and reliable data splitting even when individual SNP information is unavailable.

The application discloses a kind of light stimulation, electric stimulation, concentration gradient combined control device and preparation method, including light stimulation light intensity gradient chip layer, cell capture culture chip layer and microelectrode layer;Cell capture culture chip layer is provided with several main sample introduction sections and several capture culture channels;Capture culture channel is provided with several groups of capture culture cavities, and capture culture cavity is provided with several capture structures;Light stimulation light intensity gradient chip layer is provided with several main liquid introduction sections and several light intensity adjustment channels, and each light intensity adjustment channel is provided with light intensity adjustment section;The interdigital electrode region on microelectrode layer corresponds to the position of capture culture cavity.There is also S1-S5 etc.Preparation step.The light stimulation, electric stimulation, concentration gradient combined control device of the application can realize integrated multi-factor combined stimulation based on microfluidic platform, parallel experiment, high integration, multiple units are operated simultaneously, and has the advantages of small pollution risk, low reagent loss etc..

The invention discloses a flexible micro-nano cross-scale integrated metasurface single-cell sensor and a preparation method thereof, and relates to the field of biosensing. The sensor comprises a PDMS flexible substrate layer, a micro-nano cross-scale structure layer and a gold metal film layer, the micro-nano cross-scale structure layer adopts a nesting design of a micron-scale single cell capture unit and a nano-scale plasmon structure, so that single cell precise limiting and local electromagnetic field multi-stage enhancement are realized, and the problems of poor adaptability and low sensitivity of a traditional rigid sensor are solved. According to the preparation method, a silicon-based template-replication forming-magnetron sputtering process is adopted, a silicon template is prepared through deep ultraviolet lithography and deep reactive ion etching, a gold film is deposited after a flexible micro-nano structure is copied by PDMS, and large-scale mass production of the sensor is realized. The sensor has excellent flexibility, biocompatibility and high detection sensitivity, can realize single cell level accurate and in-situ detection, is suitable for the scenes of early disease screening, cell biological analysis, personalized medical treatment and the like, and is simple in process and low in cost.

This invention relates to the field of biomedical engineering, specifically to a device and method for high-throughput continuous sampling and gene detection of single live cells. The device includes a cell capture layer and a gene detection layer stacked sequentially from top to bottom. The cell capture layer includes a fluid inlet and a fluid outlet, and at least one capture array disposed between the fluid inlet and the fluid outlet. The at least one capture array is connected to the fluid inlet and the fluid outlet respectively via a microfluidic network. Each row of the capture array includes multiple capture units spaced apart and connected along the fluid flow direction. Each capture unit includes a main channel extending along the fluid flow direction and branch channels, the branch channels sharing the same fluid inlet and the same fluid outlet with the main channel, thereby forming a θ-shaped capture unit. The gene detection layer has multiple collection chambers spaced apart, each containing an electrode. A nanomembrane is disposed at the top of each collection chamber, and a gene detection probe is disposed at the bottom.

The invention discloses a PDMS (polydimethylsiloxane) micro-fluidic chip for long-term culture of living cells and a preparation method thereof, and relates to the technical field of micro-fluidic cell culture.The PDMS micro-fluidic chip comprises a PDMS main body, and at least one culture flow channel is arranged in the PDMS main body; the culture flow channel is provided with a fluid inlet and a fluid outlet; according to the PDMS micro-fluidic chip for long-term culture of living cells and the preparation method of the PDMS micro-fluidic chip, the mechanical microenvironment of an in-vivo tissue vasculature is effectively simulated, and meanwhile, the design of an open pool with a one-way valve at the tail end is combined, so that automatic removal of bubbles, stable maintenance of system pressure and low-damage convenient recovery of cell samples are realized; therefore, a plurality of key steps of cell capture, long-term culture, sample acquisition and the like are integrated into a simplified and reliable operation process, and the defects of complicated operation, high dependency on auxiliary equipment and insufficient microenvironment uniformity are overcome.

The invention relates to the technical field of microfluidics, and particularly discloses a cell communication micro-fluidic chip with a cell capturing structure and application of the cell communication micro-fluidic chip. The chip comprises main channels, and the two main channels are arranged in parallel at an interval; the cell interception structures are arranged in the main channels, grooves capable of accommodating at least one cell to be cultured are formed in the incident flow surfaces in the main channels, and at least one opening only allowing a culture solution to flow out is formed in each groove, so that the cells can stably stay and are uniformly distributed, and meanwhile, fluid is allowed to pass through; the culture solution interaction structure is arranged between the two main channels and used for interaction of culture solutions in the two main channels, transverse exchange of the culture solutions and cell secretions is achieved, cell migration is prevented at the same time, and therefore the non-contact cell communication model is constructed. The cell communication micro-fluidic chip can efficiently simulate the cell paracrine communication process, and is suitable for various cell communication research scenes such as inflammatory response, immune regulation, tumor microenvironment, stem cell secretion factor effect and the like.

This application relates to the field of 3D printing technology, specifically to a photomagnetic dual-drive cell capture device and a method for manufacturing the cell capture device. The cell capture device is used to capture target cells from a target sample liquid. The cell capture device includes: a capture clamp composed of multiple claw arms for capturing target cells; each claw arm is made of a magnetic composite material with predictable thermal deformation; a substrate, which is cylindrical and has a cross-sectional end face for fixing the capture clamp; the multiple claw arms are respectively connected to the cross-sectional end face of the substrate; the cross-sectional end face of the substrate is perpendicular to the axial direction of the substrate. The cell capture device provided in this application solves the problems of existing cell capture devices, such as limited functionality, narrow applicability, and easy damage to target cells, thus improving the success rate of cell capture and ease of use.

The invention relates to the field of molecular biological technology high-throughput single cell sequencing, and discloses a non-instrument-dependent high-throughput single cell library building method which comprises the following steps: S1, water-in-oil microdroplet generation; s2, temperature-controlled cell lysis: heating the water-in-oil system to activate the lysis reagent for cell lysis; s3, demulsification and microsphere cleaning: adding a demulsification buffer solution and perfluorooctanol for demulsification, and cleaning to remove invalid mRNA; s4, molecular tag release and reverse transcription; s5, amplifying and purifying the cDNA; and S6, library construction and sequencing. According to the method for generating the water-in-oil droplets without depending on microfluidic equipment, the water-in-oil droplets are generated, about 1-10% of effective droplets contain cells, molecular bar code gel beads and a cell lysis system at the same time, the polycellular rate in the effective droplets is lower than 5%, the cell capture rate is about 45%, and the method can replace the method for generating the water-in-oil droplets through the microfluidic equipment.

The application relates to the field of microfluidic chips, cell culture and analysis, in particular to an integrated microfluidic system and method for single cell capture, stimulation and analysis, wherein the integrated microfluidic system for single cell capture, stimulation and analysis is composed of two microfluidic chips in front and back parts, and the front half part and the rear half part are connected through a capillary hose c. The application realizes sorting capture, mechanical stimulation and culture analysis of single cells through an integrated pretreatment module and a capture culture module. The first microfluidic chip located at the pretreatment module makes the cell suspension flowing into the pretreatment module realize accelerated mixing under the action of the secondary flow of the bend, and the cells in the cell suspension are sorted to facilitate the next step. The second microfluidic chip located at the rear half part and composed of multiple cell capture units captures and cultures sample cells with a required size for subsequent observation and analysis.

The technical idea of the present invention provides a genetic material extraction apparatus comprising: a substrate for accommodating, on a main surface thereof, a sample in which cells are suspended; a first electrode disposed on the main surface of the substrate; a second electrode disposed to face the first electrode along a first horizontal direction, and configured to generate an electric field across the first electrode and the second electrode; a first insulating film disposed on an upper surface of the first electrode and having a first window exposing a portion of the first electrode; a second insulating film disposed on an upper surface of the second electrode and having a second window exposing a portion of the upper surface of the second electrode; and a power source for applying an alternating current voltage to the first electrode and the second electrode so as to generate the electric field for trapping the cells within the first window or the second window.

Disclosed herein include systems, methods, compositions, and kits for the generation and use of oligonucleotide-conjugated beads wherein an individual bead has a different cell label for each capture sequence. The disclosed compositions and methods enable libraries generated from each capture sequence of an oligonucleotide-conjugated bead to be identified separately bioinformatically in some embodiments.

The application discloses a kind of integrated microfluidic biological sensing chip based on micro column array D type optical fiber, belong to biological detection field.This integrated microfluidic biological sensing chip based on micro column array D type optical fiber includes fiber base micro column array D type sensing optical fiber;Periodic micro column array is arranged on the side edge polishing surface of fiber base micro column array D type sensing optical fiber;Micro column array surface is successively coated with sensitizing layer and specific modification layer;The raw material of sensitizing layer includes sensitizing material, and sensitizing material includes MXene material and / or metal nano material, and the raw material of specific modification layer includes biological aptamer material.Micro column array structure can be used as cell capture enhancement interface, forms a large number of capture sites, increases the contact area of cell and chip surface, improves cell capture efficiency.Sensitizing layer can greatly enhance the evanescent field intensity of optical fiber, and biological aptamer material realizes specific identification, and also has good sensitivity and detection accuracy to very low concentration circulating tumor cell.

The invention provides application of polyethylene glycol in reducing diffusion of cell contents in a single-cell suspension and improving single-cell capture efficiency. When the single-cell suspension containing polyethylene glycol is used for capturing single-cell nucleic acid, diffusion of contents of single cells, such as nucleic acid such as genome DNA or transcriptome RNA, can be effectively reduced, so that the contents of the single cells are more gathered, and the cell flux and capturing efficiency are effectively improved.

A droplet array integrated circuit (IC) for biological cell analysis can include a semiconductor IC substrate with a hydrophobic region disposed on a first surface thereof. An array of hydrophilic regions can be positioned on the first surface, with individual hydrophilic regions bordered by the hydrophobic region. The IC can include a plurality of biological cell-adhering elements, with individual elements disposed atop corresponding hydrophilic regions of the array. The biological cell-adhering elements can be selective for adhering to specified cell types, sizes, shapes, or morphologies, enabling precise control over cell capture and positioning.

The invention discloses a phage multifunctional affinity interface as well as a preparation method and application thereof, and belongs to the field of functional biological materials. The preparation method of the phage multifunctional affinity interface comprises the following steps: modifying a phenylboronic acid group by using pVIII protein on the side wall of an M13 phage as a bracket; connecting histidine on the protein of the M13 bacteriophage pIII to the nickel magnetic microbead; the method comprises the following steps: carrying out fluorescence labeling on a biotinylated aptamer targeting EpCAM positive cells, hybridizing with a complementary chain of a labeled fluorescence quencher, and coupling with avidin according to a molar ratio of 4: 1 to obtain an avidin-biotinylated DNA switch; and then the M13 bacteriophage-nickel magnetic microbead containing the phenylboronic acid group is coupled with an avidin-biotinylated DNA (Deoxyribose Nucleic Acid) switch. The capture efficiency of the multifunctional affinity interface to MCF-7 cells reaches 94.2%, the release efficiency reaches 92.7%, and the multifunctional affinity interface has reproducibility and can be used for multiple rounds of cell separation.

This invention discloses a single-cell capture microfluidic chip device, its fabrication method, and related components. The single-cell capture microfluidic chip device includes a glass substrate and a silicon-based cap. Grooves are formed in the glass substrate to create a main channel and side channels. Inlet and outlet are provided in the grooves. The depth of the grooves, the width of the main channel, and the width of the side channels are all greater than the diameter of the cell. The main channel and side channels are respectively located between the inlet and outlet. The main channel is a straight channel, while the side channels are non-linear channels. A capture unit protrudes from the main channel for capturing cells. The silicon-based cap is fixedly connected to the glass substrate at the opening end of the groove, and a sensing window is formed on the silicon-based cap, located above the main channel and side channels. This invention can isolate a vacuum environment, maintain cell viability, and facilitate observation and detection.

The invention discloses a mechanical phenotype analysis chip for screening bacterial infected circulating tumor cells, and belongs to application of a micro-fluidic chip technology in the field of biomedical in-vitro diagnosis. The system comprises a micro-fluidic chip and a detection solution. The micro-fluidic chip comprises a sample introduction end, a shunting flow channel, a main flow channel and a sample discharge end, wherein single-cell mechanical property analysis function unit arrays are uniformly distributed on the two sides of the main flow channel in a staggered manner; and the single cell mechanical property analysis functional unit comprises a hook-shaped structure for single cell anchoring and a deformation channel structure for single cell mechanical property analysis. The detection solution comprises a fluorescent probe capable of specifically recognizing bacteria in cells. According to the micro-fluidic chip disclosed by the invention, single cell capture, mechanical property analysis and intracellular bacteria detection can be integrated on the same chip, and a time-saving, low-cost and high-throughput single cell detection platform can be constructed, so that visual detection of single cell mechanical phenotypes and intracellular bacteria is realized; the method can be used for evaluating the metastasis potential of the circulating tumor cells in the bacterial infection breast cancer.

A microfluidic control chip, system, and method for single-cell extracellular vesicle isolation and analysis are disclosed. The microfluidic control chip includes a flow layer and a control layer. The flow layer includes a sample / air inlet, a waste liquid outlet, a micron-sized trapping channel containing contraction and expansion structures, a micron-sized gas flow channel, a micron-sized single-cell trapping structure, and a micron-sized single microsphere trapping structure. The control layer includes an air inlet, a micron-sized gas flow channel, and a micron-sized protrusion structure. A microfluidic control chip imaging system for single-cell extracellular vesicle isolation and analysis is also disclosed, comprising the microfluidic control chip, a micro-injection pump, a pressure controller, a capillary tube, a PFA tube, and a fluorescence microscope. The microfluidic control chip of this invention can simultaneously analyze biomarkers expressed on the surface of single cells and single-cell secretory extracellular vesicles. Correlation analysis of cancer cell drug resistance with the expression levels of biomarker proteins on the surface of cells and cell secretory extracellular vesicles is achieved at single-cell resolution.

Ballistic microscopy—a completely new approach to “image” a cell utilizing particle bombardment, is described. These are ballistic micro and nano particles that travel through a cell at ballistic speed and capture a pico or femto-liter of cellular content and bring it out for analysis without harming the cell. This enables a new approach to omics-based imaging where millions of these particles are bombarded on cells with resolved space and time and captured to process using well known omics techniques including proteomics (mass spec) or sequencing—while keeping the spatial and temporal resolution. This work provides—for the first time—a way to resolve atomic details of live cells without any labels.

The invention discloses an in-vitro culture method of histolytic amoeba, and relates to the technical field of in-vitro culture of microorganisms, and the in-vitro culture method comprises the following steps: S1, preparing a semisolid agar basic culture medium containing a fructus forsythiae alcohol extract to obtain a basic culture system; s2, obtaining a chip culture system; s3, obtaining a dynamic culture; s4, obtaining an optimized culture; s5, obtaining a pure-line histolytic amoeba culture; according to the invention, a single-cell capture structure focused by fluid power of the micro-fluidic chip is utilized to realize accurate capture and separation of single insect bodies without damage and with high flux: the fluid focusing effect in the chip can limit the insect bodies in independent separation chambers one by one, and the initial purity of pure line culture is ensured from the source; meanwhile, due to integrated culture in the chip, environmental exposure and mechanical damage in a polypide transferring process are reduced, the activity of the separated polypide is maintained, and the problems of low purity, large operation damage and low efficiency of a traditional separation method are solved.

A single-cell capture method, comprising the following steps: by means of a fluid, injecting single cells to be captured into a micropore-coupled microchamber microfluidic chip, wherein the chip comprises a substrate and a main channel provided thereon, the surface of the main channel is provided with a plurality of micropores and microchambers corresponding to the micropores, and the microchambers are in communication with the main channel; after the fluid fills the main channel, keeping the fluid in the main channel stationary, so that the single cells fall into the micropores under gravity; after the micropores capture the single cells, introducing the fluid again to discharge the uncaptured single cells; and after discharge, inverting and tilting the chip, so that the single cells enter the microchambers. The method solves the technical problems in the prior art of low single-cell capture efficiency, insufficient capture precision, the need for complex external devices for control, and being unable to complete single-cell capture, culture, and other operations on the same chip.

The present invention provides devices and methods for single-cell capture and analysis from complex solutions. The method includes the preparation of substrates for the capture of single cells for multiplexed analysis.

The invention discloses a functional modified nanoneedle chip as well as a preparation method and application thereof, and relates to the technical field of biological chips. Specifically, the functional modified nanoneedle chip disclosed by the invention is obtained by sequentially modifying a nanoneedle chip with NHS-biotin, streptavidin and biotin-poly (T), wherein the length of the poly (T) of the biotin-poly (T) is 20 to 30 nt. According to the method, a three-step modification method of NHS-biotin, streptavidin and biotin-poly (T) is adopted, a functionalized modified nanoneedle chip treatment system is optimized, the extraction efficiency of 106-level cell RNA is larger than or equal to 85%, the cDNA concentration reaches 780 ng / mu L or above and is improved by 27% compared with that of an unmodified nanoneedle group, the problems that 106-level cell RNA capture is insufficient and time consumption is long are effectively solved, and the method has important application prospects.

The invention relates to a micro-fluidic chip for specific single cell screening and a use method of the micro-fluidic chip. The micro-fluidic chip comprises four layers, namely a detection layer, a thin film layer, a pump layer and a substrate layer in sequence from top to bottom, and the detection layer comprises a capture channel, a sample introduction channel and a lantern-shaped microcavity array and is used for capturing and imaging detection of single cells; the thin film layer is arranged below the detection layer and forms a closed space with the microstructure on the detection layer; the pump layer comprises a pump control channel and a rectangular microstructure array, and the rectangular microstructures are arranged below the lantern-shaped microcavities in a one-to-one correspondence mode during assembly. When cell screening is carried out, firstly, sample cells, detection liquid, substrate liquid and the like are sequentially introduced into the detection layer, and single cell capture, isolation, incubation reaction and array imaging detection are carried out; and then, selectively applying pressure to the rectangular microstructure under the lantern-shaped microcavity corresponding to the selected specific cells, so that the thin film layer in the corresponding area is quickly deformed, and further selective release and recovery of the cells are realized. The chip can realize single cell detection and addressable noninvasive release, is simple to operate and small in cell damage, and has a good application prospect in the field of single cell analysis and screening.

The invention relates to a micro-fluidic chip for heterogeneous cell electrofusion, which is characterized in that a partition wall and two micro-channels are arranged in the chip, the two micro-channels are bilaterally symmetrical by taking the partition wall as a symmetry axis, and each micro-channel is provided with a main channel and a bypass channel; a plurality of capturing cavities are formed in the main channel; the bypass channel is shunted from the main channel at the inlet of each capturing cavity, extends towards the direction far away from the main channel and then turns back in a detour manner, and the bypass channel and the main channel are converged at the outlet of the capturing cavity; a fluid resistance difference exists between the main channel and the bypass channel, so that a cell suspension preferentially enters the capturing cavity of the main channel to capture a single cell when entering; the capturing cavities which are symmetrical left and right are communicated through a pairing channel, so that the two captured cells are in close contact and are fused under the action of a pulse signal to form a binuclear heterogeneous cell. Through special flow channel geometric design and fluid resistance matching, efficient and accurate pairing and electric fusion of heterogeneous cells are realized, and the method has the advantages of high flux, good fusion rate, less non-specific fusion and the like.