102 results about "Droplet microfluidics" patented technology

Methods of utilizing magnetic particles or beads (MBs) in droplet-based (or digital) microfluidics are disclosed. The methods may be used in enrichment or separation processes. A first method employs the droplet meniscus to assist in the magnetic collection and positioning of MBs during droplet microfluidic operations. The sweeping movement of the meniscus lifts the MBs off the solid surface and frees them from various surface forces acting on the MBs. A second method uses chemical additives to reduce the adhesion of MBs to surfaces. Both methods allow the MBs on a solid surface to be effectively moved by magnetic force. Droplets may be driven by various methods or techniques including, for example, electrowetting, electrostatic, electromechanical, electrophoretic, dielectrophoretic, electroosmotic, thermocapillary, surface acoustic, and pressure.

The invention provides a novel droplet micro-fluidic chip and an operation method thereof, and in particular relates to a droplet micro-fluidic chip for integrating an electroosmotic pump. The droplet chip consists of a sample pool, a micro-channel and a micro-electroosmotic pump. One or the combination of micro-droplet generation, splitting or fusion is controlled by regulating negative pressure generated by the micro-electroosmotic pump. Droplet operation is realized in a non-electric field environment. Under the condition, on one hand, droplets can be accurately operated by regulating the pressure of the electroosmotic pump; on the other hand, the droplet generation, splitting and fusion conditions are mild, and interference from an additional electric field is avoided.

An integrated liquid drop microfluidic chip comprises one or more liquid drop generation structures, every liquid drop generation structure is connected with a corresponding collection cavity structure through a corresponding connection channel, a water phase reagent and an oil phase reagent respectively go through the one or more liquid drop generation structures to generate liquid drops with uniform dimension, an emulsion including the liquid drops goes through the connection channel and directly or indirectly enters the collection cavity structure, and is adaptively arranged to form a regular liquid drop array in the collection cavity structure, the liquid drop array undergoes a biochemical reaction under constant temperature control or temperature circulation control in an air heating refrigeration or Peltier semiconductor heating refrigeration mode, and real time monitoring or timed optical detection is also carried out. The integrated liquid drop microfluidic chip reduces the sample transfer process, can be conveniently integrated with a thermometric module and an optical detection module, greatly improves the reaction and detection speed, and also simplifies the complex degree of a whole apparatus system.

The invention relates to a high-throughput detection system for microbes based on a droplet microfluidic chip. The system mainly comprises a droplet microfluidic chip system (1), a light path system (2) and a data acquisition and analysis system (3), wherein the droplet microfluidic chip system (1) embeds to-be-detected microbe to an independent single droplet micro-reaction chamber; the light path system (2) is used for carrying out laser-induced fluorescence detection signal transmission of the microbe sample in the single droplet micro-reaction chamber; and the data acquisition and analysis system (3) carries out detection and analysis on the acquired signals through a computer software. The droplet microfluidic chip can be replaced. The system is suitable for laser-induced fluorescence detection and analysis of different types of microbes, has the advantages of simple design, low processing and manufacturing cost, low detection cost, fast speed and high throughput, and realizes efficient screening of production-related target enzymes and metabolites of different types of industrial microbes including escherichia coli, corynebacterium glutamicum and saccharomyces cerevisiae.

A microfluidics droplet detection system based on a capacitive sensor is characterized by comprising a droplet microfluidics chip, a direct-current voltage stabilizing circuit unit, a signal detection circuit unit and a computer, and the droplet microfluidics chip is of a structure according to which the capacitive sensor is integrated with a microchannel used for generating micro-droplets. A droplet detection method comprises the steps of sample pumping, droplet formation and droplet detection. The system and method have the advantages that the structure is simple, detection is conducted by means of the change of the capacitance, detection is quick, the size of the system is small, integration can be realized, the system and method are suitable for analysis of expensive samples and reagents, and manufacturing and implementation are easy.

The invention relates to a system for rapidly, directly, absolutely and quantitatively detecting microbes based on a liquid drop by using a liquid drop micro-fluidic technology, and a method thereof. The system comprises a micro-fluidic chip, a plurality of cell collecting, culturing and observing chambers, an optical detection unit and an image processing unit. The method comprises the following steps: preprocessing a sample, injecting the preprocessed sample into the micro-fluidic chip to form monodispersed unicellular liquid drops, and carrying out controlled temperature cell culture on the monodispersed unicellular liquid drops formed in the cell collecting, culturing and observing chambers for 1-2h; and determining the existence of living cells and the growth speed of the cells according to the turbidity of a single liquid drop, collecting the amplified pictures of the liquid drops in the cell collecting, culturing and observing chambers, and counting the turbid liquid drops to obtain the quantity of the microbes in the sample. The system for rapidly, directly, absolutely and quantitatively detecting microbes based on a liquid drop and the method have the advantages of simplicity and low cost, are suitable for portable onsite detection and laboratory detection, and can be used to detect microbe indexes in the fields of foods, environment, drugs sanitation products and cosmetics.

The invention provides a single cell separation method based on droplet micro-fluidic chips. The method concretely comprises the following steps: A, a single cell suspension flows into a dispersed phase inlet channel from a dispersed phase inlet; B, an oil phase liquid flows into a continuous phase inlet channel from a continuous phase inlet; C, the above two phases merge to form droplets enclosing single cells, the droplets flow over a droplet capturing unit with the generation of a large number of droplets in a liquid storage pool, and stand for 2-5 min, and the superfluous droplets are sucked out when the droplets slowly settle into the droplet capture unit; and D, the droplet chips which capture the single cells are cultured in a 37 DEG C incubator, then DAPI is added to carry out nuclear staining, and the single cell capture rate is detected. The method has the advantages of simplicity and rapidity in operation, small use amounts of the cells and reagents, low experiment cost, high integration and wide application range.

In alternative embodiments, the invention provides high throughput, multiplexed systems or methods for detecting a biological, a physiological or a pathological maker, or a single molecule or a single cell using a droplet microfluidics system integrated with use of a sensor or a sensing system, an aptamer, or a DNAzyme. In alternative embodiments, the sensor or sensing system comprises a nucleic acid based, an antibody based, an enzyme based or a chemical based sensor or sensing system. In alternative embodiments, the invention provides methods for detecting a biological, a physiological or a pathological marker, or a single molecule or a single cell using a droplet system integrated with rapid and sensitive fluorescence detection systems including, for example, a 3D particle detector. In alternative embodiments, the invention provides systems comprising integrated comprehensive droplet digitial detection (IC 3D).

The invention provides an integrated droplet microfluidic chip structure. All functional modules of droplet generation, amplification and detection are integrated on the same microfluidic chip to achieve the whole enclosed process from the droplet generation to fluorescence detection. The invention also relates to an integrated droplet microfluidic chip structure preparation method and a microfluidic chip assembly. The structure is compatible with the positive pressure or negative pressure drive mode, the pressure response time is short, the rapid droplet generation can be achieved, and the sample preparation time is greatly reduced. Droplet generation oil is not required to be filled in advance, the operation is simple, and popularization and application in the technical field of digitalPCR are facilitated.

The invention belongs to the technical field of preparation of phase change microcapsule materials, and discloses a double emulsified glass capillary microfluidic chip and a phase change microcapsuleprepared thereby. The double emulsified glass capillary microfluidic chip is used to prepare a phase change material microemulsion, the controllability of the prepared capsule is greatly enhanced, andthe practical value of the capsule is increased; and a high precision syringe pump is used to precisely control the emulsification process of a core material and a wall material in the microfluidic chip, and the phase change material microemulsion with good monodispersity and high sphericity is obtained. An UV curing wall material is used to reduce preparation time and cost, and the controllability of the wall material is improved. The preparation process has the advantages of simple equipment, simple operation, high utilization rate of raw materials, easy control of product particle size, sphericity, small shell thickness and core material size, no noise, no harmful waste, low energy consumption, and the like, and is suitable for promotion and application of various phase change microcapsules in scientific research and industry.

The invention relates to a preparation method for a spherical cavity equipped polymer fiber and a special microfluidic chip. With the chip and the method, polymer fiber with a diameter ranging from micron level to millimeter level can be prepared. Spacing and size controllable spherical cavities can continuously be formed in the polymer fiber, and various inclusions can be loaded in the cavities. The method combines a droplet microfluidic technology and a spinning microfluidic technology, utilizes the droplet microfluidic technology to disperse a volatile liquid uniformly in the continuous flow of a polymer monomer solution in the form of droplets, and utilizes the spinning microfluidic technology to form polymer fiber and arrange the droplets therein orderly. The droplet solution is removed from solidified polymer fiber by means of its volatilization characteristic so as to form the spherical cavities with uniform and controllable size and adjustable spacing. The substances added into the droplets in advance can be loaded into the fiber cavities, and the material can be endowed with flexible performance so as to be expected for application in wide fields.

The invention provides a liquid-drop microfluidic chip for separation of single cells and a preparation method for the liquid-drop microfluidic chip. The chip consists of two layers, i.e., an upper layer and a lower layer, wherein the upper layer is a liquid drop generating unit, and the lower layer is a liquid drop trapping unit; and the liquid drop generating unit is provided with structures asfollows: a dispersed phase inlet (1), a continuous phase inlet (2), a dispersed-phase liquid inlet passage (3), a continuous-phase liquid inlet passage (4), a liquid drop generating cross passage (5),a liquid outflow passage (6), a liquid outlet (7) and a liquid storage tank (8). According to the chip, an SU-8 chip template with partial-bulging passages is prepared by adopting a photoetching andcorroding method and is subjected to stripping by polydimethylsiloxane (PDMS), thereby obtaining a PDMS chip. The chip has the flexible liquid drop generating unit and has the high-pass single-cell trapping unit; and the chip is simple in structure, convenient in operation, high in efficiency, low in consumption of cells and reagents and extensive in range of application.

The invention discloses a droplet microfluidic technology based method for preparing chromatographic packing with the uniform and controllable grain size, and relates to preparation of the chromatographic packing. Tetramethoxysilane, 3-glycidoxypropyltrimethoxysilane and polyethylene glycol are added into an acetic acid solution and hydrolyzed into a hydrolysate under an ice-water bath, and a water-soluble basic amino acid is added and subjected to ultrasound until complete dissolution to be used as a disperse phase; an oil-soluble surfactant is dissolved into oil to be used as a continuous phase; the disperse phase is introduced into a horizontal channel of a microfluidic chip, the continuous phase is introduced into a vertical channel of the microfluidic chip, and the ratio of flow velocities of the disperse phase and the continuous phase is adjusted at the interface of the horizontal channel and the vertical channel to produce droplets with different grain sizes; the droplets have a condensation reaction at the temperature of 30-50 DEG C for 10-14 h and have an epoxide ring-opening reaction at the temperature of 60-80 DEG C for 10-14 h respectively, then isopropanol and methyl alcohol aqueous solutions are used for alternative washing respectively, and a product is obtained through vacuum drying.

The invention discloses a lactoflavin engineering bacterium and a constructing method thereof, and the preservation number of the lactoflavin engineering bacterium is CGMCC NO.16132. Through a geneticengineering means, a lactoflavin auxotroph strain is constructed, then a chromosome integrates and expresses a lactoflavin operon demodulated by bacillus amyloliquefaciens, and the lactoflavin production capacity of the strain is restored; then an expression vector overexpresses the lactoflavin operon demodulated by the bacillus subtilis, so that the lactoflavin production capacity of the straincan be improved; in addition, through point mutation of a ribC gene, the catabolism of lactoflavin is diminished; and finally, a recA gene which is responsible for homologous recombination and DNA repair on the chromosome is inserted for inactivation, and a lactoflavin production chassis cell is constructed. Based on the lactoflavin production chassis cell, through combination of the characteristic that the lactoflavin is a natural flroresence substance, and a strain high in lactoflavin yield is screened out through liquid drop micro-fluidic control. The invention also discloses a method for producing the lactoflavin through fermentation with the bacillus subtilis, and an application of the strain high in lactoflavin yield to lactoflavin production and an application of the strain high inlactoflavin yield to medicines, foods and feeds.

The present invention provides a liquid drop microfluidic chip including a chip substrate, a chip channel layer, a liquid drop storage layer, a chip adhesive layer and a chip cover plate, the chip channel layer, the liquid drop storage layer, the chip adhesive layer and the chip cover plate are sequentially pressed and sealed with the chip substrate, the liquid drop storage layer and the chip adhesive layer are respectively correspondingly provided with a liquid drop channel, the opposite two ends of the liquid drop channel are respectively provided with a liquid drop inlet and a liquid drop outlet, the chip channel layer is also provided with a liquid drop generation component and a liquid drop detection / separation component, the liquid drop generation component is in pipe connection with the liquid drop inlet of the liquid drop channel, the liquid drop detection / separation component is in pipe connection with the liquid drop outlet of the liquid drop channel. The liquid drop microfluidic chip has integrated many functions such as liquid drop generation, storage, reaction and sorting, has a high integration degree and a small area, and can achieve high flux separation experiment under high screening efficiency and automation level. The invention also provides a preparation method of the chip, and the preparation method is suitable for preparation of the chip in a large scale and simple way.

The invention discloses a droplet PCR amplification detection device based on a microfluidic chip. The device comprises the droplet microfluidic chip, an XYZ motion unit, a PCR amplification unit anda detection unit. A droplet containing a DNA molecule is introduced into the droplet microfluidic chip through a pipette after being generated, the chip is placed in the droplet PCR amplification unitto perform a PCR amplification reaction, and fluorochrome in the droplet is motivated through an LED lamp and light filter in the droplet detection unit, a camera is controlled to take a picture of the fluorescent droplet, the picture of the fluorescent droplet is focused through an autofocus algorithm and the control of a Z motion platform, and acquiring of images of different portions of the chip is completed through chip marking and the control of an XY motion platform; the captured images are spliced to obtain complete information of the chip droplet region images, thereby completing droplet PCR amplification detection. The device has the advantages that the operation is simple, the reagent consumption is low, the droplet stability is good, the detection accuracy is high, and the PCRamplification diagnosis and analysis can be efficiently completed.

The invention belongs to the field of micro-fluidic chips, and relates to a biological macromolecule detection method. An alpha-LISA technology and a droplet-based microfluidics technology are combined and optimized to prepare a micro-fluidic chip liquid. A novel biological macromolecule detection method with a high sensitivity is provided. A three-phase mixing technology is adopted. A passive mixing structure is used to fully and evenly mix the reaction liquid. A cross focusing technology is used to form droplets. The generated droplets have the advantage of smaller volume, and thus the reactions become faster. Moreover, the requirements on the samples are lower, the reactions are specific, sensitive, fast, and full; the washing does not need to be carried out after reactions; complicated complexes such as complete proteins, enzyme complex, bacteriophage, and the like, can be detected; the technical bottleneck of conventional commercial liquid chips at present can be broken through; the provided method can be applied to fast clinical biological macromolecule detection; the detection sensitivity and specificity are both improved, and the required sample amount is reduced.

The invention relates to a liquid drop micro-fluidic chip based on a microlens array. The liquid drop micro-fluidic chip comprises a chip body, a reflection membrane layer, a light transmission plate and a microlens, wherein at least one flow channel is formed in the surface of the chip body; the reflection membrane layer is arranged on an inner wall of the flow channel; the light transmission plate covers the flow channel; and the microlens is arranged on the light transmission plate and is located just above the flow channel. By using a light energy gathering property of the microlens array, a microlens array technology is combined with the liquid drop micro-fluidic chip, so that the intensity of fluorescent light received by a detector can be extremely enhanced; and by optimally designing a liquid drop flowing pipeline of a detection region and improving the quantity of parallel detection channels, the detection flux can be improved by 1-2 orders of magnitudes as compared with that of a traditional flow type technology.

The invention provides a high-throughput screening apparatus using a droplet micro-fluidic chip. The apparatus comprises: a rack; a micro-fluidic control system constructed to control a fluid supply device for supplying a droplet-containing fluid to the droplet micro-fluidic chip; a photoelectric detection system comprising an incident light generation device for limiting outgoing and incident light paths, a preferable laser excitation device and a fluorescence detection device for limiting a fluorescent light path; an electric screening system constructed to connect the droplet micro-fluidicchip in order to screen droplets flowing through the droplet micro-fluidic chip; and one or more controllers constructed to receive a signal from the fluorescence detection device, compare the signalwith a predetermined threshold and control the electric screening system to perform droplet screening according to the obtained comparison result.

The invention relates to a liquid drop microfluidic system for detecting interaction between quantum dots and biomolecules. The liquid drop microfluidic system comprises a microfluidic chip, a fluorescence microscope, three injection pumps for injecting liquid drops to the microfluidic chip, a fluorescence interface, a fluorescence spectrograph and a data acquiring and analyzing system, wherein the microfluidic chip is arranged on the focal point of an objective lens; one end of the fluorescence interface is connected with a standard port of the fluorescence microscope, and the other end of the fluorescence interface is connected with the fluorescence spectrograph through an optical fiber; and a spectral signal output end of the fluorescence spectrograph is connected with a signal input end of the data acquiring and analyzing system. The invention also relates to a method for detecting the interaction between the quantum dots and the biomolecules, the biomolecules are marked by dye molecules, and fluorescence resonance energy can be transferred between the dye molecules and the quantum dots. The liquid drop microfluidic system is easy to operate and high in detection sensitivity, and rapid dynamic detection between the quantum dots and the biomolecules can be realized; and the fluorescence spectrograph can simultaneously detect a plurality of fluorescence wavelengths, errors are reduced, and detection accuracy is improved.

The invention provides a preparation method of an encapsulated phase change material with high thermal conductivity coefficient and uniform particle size. Metal particle doping and droplet microfluidic technologies are employed for a one-step emulsification and encapsulation process of a phase change material in a microfluidic chip. The phase change material in the capsule is doped with metal nanoparticles with high thermal conductivity coefficient, thus effectively increasing the thermal conductivity coefficient of the capsule and strengthening the heat transfer performance of the encapsulated phase change material. At the same time, the latent heat of phase change in the capsule has little reduction, and the practical value of the capsule is increased. The method provided by the invention uses droplet microfluidic technology to prepare a phase change material microemulsion, and uses a high precision injection pump to precisely control the emulsification process of droplets, thus obtaining a monodisperse phase change material microemulsion with narrow particle size distribution. The preparation process has the advantages of simple equipment, easy operation, little waste of raw materials, easy control of particle size, no noise pollution, and low energy consumption, etc., and is suitable for popularization and application in scientific research and industrial production of encapsulated phase change materials.

The invention relates to a liquid drop micro-fluidic chip used for detecting the diabetes with high sensitivity and a detection method, and belongs to the technical field of microfluidics. The liquiddrop micro-fluidic chip sequentially comprises a cover glass layer 20, an upper fluid passageway layer 30, a lower fluid passageway layer 40 and a base layer 50; and a liquid drop micro-fluidic chip based high sensitivity diabetes detection device comprises a high pressure liquid phase chromatographic pump 2, a sample inlet valve 3, the liquid drop micro-fluidic chip 6, a deuterium lamp and a halogen tungsten lamp source 10, an ultraviolet-visible light spectrometer 11 and data processing software 13. The provided micro-fluidic chip is small in size, low in cost and simple in operation, and isfavorable for real-time detection on the site. The high sensitivity diabetes detection method is not only suitable for the high sensitivity detection of a sample HbA1c, but also suitable for detection of biomacromolecules like polypeptide, nucleic acid and protein.

The invention discloses a high-throughput single-cell transcriptome sequencing method and a kit. The high-throughput single-cell transcriptome sequencing method includes the steps that a droplet generation system is adopted to encapsulate a single cell and labeled microbeads in a droplet, and reverse transcription is performed in the droplet. According to the high-throughput single-cell transcriptome sequencing method, a throughput of 9,000 cells which is equivalent to 10 x genomics can be achieved, after the droplet is demulsified, the microbeads are less contaminated with each other, and theproportion of valid data is increased. According to the high-throughput single-cell transcriptome sequencing method, reverse transcription is performed in the droplet, fewer reagents are required, and the cost is low. In the preferred scheme of the high-throughput single-cell transcriptome sequencing method, a SMART template conversion technology is adopted in reverse transcription, and productscan be directly used for subsequent Tn5 library construction and BGISeq-500 platform sequencing without library conversion; and multiple amplification and introduction of deviations are avoided, a droplet-based microfluidics platform is used in conjunction with a BGISeq-500 sequencing platform, and large-scale single-cell sequencing is simplified and facilitated.

The invention relates to a centrifugal liquid drop microfluidic chip. The centrifugal liquid drop microfluidic chip comprises a substrate and a cover plate arranged on the substrate, wherein the coverplate comprises an oil phase storage cavity, a water phase storage cavity, a liquid drop generating structure connected to the oil phase storage cavity and the water phase storage cavity, a liquid drop storage cavity connected to the liquid drop generating structure and provided with at least one oil storage structure; the oil storage structure has at least one opening; and the distance from theoil phase storage cavity and the water phase storage cavity to a centrifugal shaft is less than the distance from the liquid drop storage cavity to the centrifugal shaft. The centrifugal liquid drop microfluidic chip reduces the fraction of the average dispersed phase by adjustment through the oil storage structure, improves the stability of liquid drops, and ensures that the liquid drops in a centrifugal force chip do not fuse in the storage and heating process, so that the chip has a high application value in the research application fields of biology, chemistry, medical diagnosis and the like.

The present invention provides an automatic high-throughput single cell capture method based on a liquid droplet microfluidic chip, wherein the microfluidic chip comprises two layers, the upper layeris a flow path inlet and outlet layer, the lower layer is a flow path control layer, the flow path inlet and outlet layer has a liquid flow path channel inlet and a liquid flow path channel outlet, and the flow path control layer comprises a single cell capture flow path channel, gas path channels and liquid droplet generation units. According to the present invention, by introducing the gas pressure controllable gas flow path channel, the negative pressure flow path channel can be formed and can automatically suck the single cell suspension into the capture trap so as to conveniently observeand detect the proliferation, the differentiation, the drug reaction and other behaviors of the single cells; the automatic high-throughput single cell capture is achieved by using the liquid dropletmicrofluidic technology and the fluid mechanics principle; and compared to the traditional single cell capture method, the method of the present invention has advantages of simple, convenient and flexible operation, high throughput, no pollution, wide application range, strong scalability and the like.

The invention relates to a droplet micro-fluidic chip and a preparation method of micro-droplets. The droplet micro-fluidic chip comprises at least one droplet preparation unit; the droplet preparation unit comprises a dispersed phase cavity, a quantitative cavity, a capillary nozzle and a continuous phase cavity; the droplet micro-fluidic chip is provided with a rotating center, the dispersed phase cavity is provided with a sample adding hole for adding dispersed phase liquid, and the quantitative cavity is connected with the dispersed phase cavity and is further away from the rotating centerrelative to the dispersed phase cavity; the capillary nozzle is further away from the rotating center relative to the quantitative cavity, and one end of the capillary nozzle is connected with the quantitative cavity and extends from the connecting end to the direction away from the rotating center; the continuous phase cavity is connected with the end, away from the quantitative cavity, of the capillary nozzle and is further away from the rotating center relative to the capillary nozzle, and continuous phase liquid is contained in the continuous phase cavity. According to the droplet micro-fluidic chip, centrifugal force is used as droplet preparation driving force, and stable and high-speed preparation of droplets with uniform sizes can be realized through configuration of different parameters.

A method of determining the result of an assay in a microfluidic device includes the steps of: dispensing a sample droplet onto a first portion of an electrode array of the microfluidic device; dispensing a reagent droplet onto a second portion of the electrode array of the microfluidic device; controlling actuation voltages applied to the electrode array to mix the sample droplet and the reagentdroplet into a product droplet; sensing a dynamic property of the product droplet; and determining an assay of the sample droplet based on the sensed dynamic property. The dynamic property is a physical property of the product droplet that influences a transport property of the product droplet on the electrode array. Example dynamic properties of the product droplet include the moveable state, split-able state, and viscosity based on droplet properties. The method may be used to perform an amoebocyte lysate (LAL) assay.

The invention discloses a honeycomb-shaped carbon nano tube porous microsphere, and a preparation method and application thereof. The honeycomb-shaped carbon nano tube porous microsphere is a three-dimensional porous structure formed by carbon nano tubes, has a diameter in a range of 5-200 microns, and contains a large amount of micropores with diameters in a range of of 20-1000 nm. A droplet micro-fluidic technology is adopted, and a porous microsphere structure is constructed by using micro droplets as soft templates and using SiO2 nano particle as hard templates. The honeycomb-shaped carbon nanotube porous microspheres prepared by the method are uniform in size, and the size, shape and density of ''honeycombs'' are controllable. Meanwhile, the preparation method disclosed by the invention is simple, and preparation is flexible and convenient. The flow speed is controlled through an injection pump, so that the advantages of accurate operation control, easy industrial batch production and the like are achieved.