50 results about "Intact cell" patented technology

Provided herein are novel stabiligases for use in cell membrane proteome analysis. The subject stabiligases are capable of attaching to glycans found on the surface of cell membranes to form glycan-tethered (GT) stabiligases. Such glycan-tethered stabiligases are capable of robustly and selectively attaching label probes to cell surface proteins of intact cells. The subject novel stabiligases described herein advantageously allow for the identification and profiling of cell membrane proteins that have undergone an extracellular N-terminal proteolytic event.

The invention relates in one aspect to a method for detecting an interaction between a ligand and at least one target protein in in-tact cells, comprising inducing protein precipitation using one or more solvents in a first sample and in a second sample, both comprising intact cells and / or permeabilized cells, wherein said first sample is treated with said ligand, and said second sample is not treated with said ligand, wherein a difference in tolerance of the target protein to solvent-induced precipitation between the first and second samples indicates an interaction between a ligand and a target protein.

The invention relates to the technical field of cell classification, in particular to a full-automatic classification detection method for peripheral blood cell morphology. The method comprises the following steps: acquiring a cell region in a cell display image of peripheral blood; obtaining an overlapped cell region according to the form of the cell region and the curvature of edge pixel points in the cell region; according to the length of each edge line in the overlapped cell area, the curvature of edge pixel points on each edge line and the curvature of intersection point pixel points of each edge line and other edge lines, obtaining exposed edge lines of the covered cells; and determining a complete cell edge line of the covered cell according to the curvature and the end point of the exposed edge line, and carrying out full-automatic classification detection on the peripheral blood cell morphology. By acquiring the complete cell edge line of the covered cell, the form of the covered cell is determined, the situation of inaccurate automatic identification and classification of the cell due to cell overlapping is avoided, and the accuracy of full-automatic classification detection of the form of the peripheral blood cell is improved.

This invention relates to a biomimetic nanodelivery system ANG-2-CMLNPs. The preparation method of ANG-2-CMLNPs includes the following steps: Step 1: Prepare EZ NPs using a one-step self-assembly method, then co-incubate them with siRNA to obtain siRNA-EZ NPs; Step 2: Prepare liposomes using a thin-film hydration method, then modify them with ANG-2; Step 3: Add U87 MG cells to cold PBS, centrifuge to remove intact cells and debris, and the resulting precipitate is the glioma cell membrane fusion membrane; Step 4: Mix the Angiopep-2 modified liposomes obtained in Step 2 with the glioma cell membrane fusion membrane obtained in Step 3, and sonicate to obtain a composite; Step 5: Co-extrude the composite obtained in Step 4 with the siRNA-EZ NPs obtained in Step 1 to obtain the biomimetic nanodelivery system ANG-2-CMLNPs; In the above preparation process, when EZ... When the mass ratio of NPs to siRNA is 125:1 and the mass ratio of ANG-2 modified liposomes to CM is 1:5, the prepared ANG-2-CMLNPs delivery system achieves the optimal functional balance between the ability to cross the blood-brain barrier (BBB) ​​and tumor targeting efficacy.

Provided herein are methods of isolating edible biomass from a suspension cell culture comprising metazoan cells, methods of isolating intact cells from a cell culture, cell-based food products comprising edible biomass from suspension cell cultures isolated according to the methods comprising isoelectric precipitation as described herein.

The invention relates to a method of enriching a cell population containing intact and damaged cells in a liquid medium, comprising the following steps: a.) providing the liquid medium containing the cell population, b.) sedimenting the cell population out of the liquid medium, c.) separating the liquid medium and the sedimented cell population, d.) resuspending the sedimented cell population in a suspension solution to produce a cell suspension, e.) admixing the cell suspension with a ferromagnetic material that binds specifically to the damaged cells, f.) incubating the cell suspension admixed with the ferromagnetic material, g.) introducing the cell suspension into a magnetic field in order to separate the bound cells and the unbound cells in the cell suspension, and h.) separating a fraction of the cell suspension enriched with unbound intact cells, to the use of intact sperm cells enriched in the method, and to an apparatus for separating cells bound to magnetic particles in a cell suspension.

The present disclosure is directed to a non-enzymatic dissociation method which facilitates the dissociation of one or more formalin-fixed paraffin-embedded tissue samples into dissociated single cells. The present disclosure is also directed to methods of single cell analysis, i.e., methods of analyzing and / or measuring target components (e.g., biomolecules such as, but not limited to, polypeptides, polynucleotides, small molecules, and the like) on or in cells non-enzymatically dissociated from one or more FFPE tissue samples.

Disclosed are methods for isolating an analyte from a sample. In some aspects, the methods are for selectively isolating a microbial cell analyte, such as a nucleic acid, from a sample containing or suspected of containing mammalian cells. The selective isolation method includes selective lysis of the mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analyte. The released analyte is then eluted from the filter. In other aspects, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes may be analyzed using a suitable assay depending on the type of analyte molecule. Also disclosed are fluidic systems and lysis solutions that may be used for isolating an analyte according to the disclosed methods.

Methods for isolating analytes from a sample are disclosed. In some embodiments, the methods are for selectively isolating microbial cell analytes, such as nucleic acids, from a sample containing or suspected of containing mammalian cells. The selective isolation method involves selective lysis of mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analytes. The released analytes are then eluted from the filter. In other embodiments, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes can be analyzed using an appropriate assay depending on the type of analyte molecule.

Compositions and methods for analysis of protein expression at the single cell level are described. The single cell proteomic technology described herein implements peptide barcode molecules to uniquely label all of the proteins and / or polypeptides present in a single cell, whereby all of the proteins / polypeptides (proteome) present in a single intact cell are uniquely labeled with a peptide barcode molecule in a partition. Polypeptides from a single cell can be uniquely barcoded and via protein sequencing, such as nanopore sequencing. Proteomic analysis of the protein sequencing data elucidates the entire proteome that was expressed in the single cell.

The invention discloses a cell fusion degree auxiliary verification method and apparatus, and a server. Stem cell microscope images are collected by using three set resolution scales of an initial resolution, a down-sampling resolution and an up-sampling resolution to form a corresponding standard image group, a low-resolution image group and a high-resolution image group; and performing different image processing on different resolutions, performing cross-scale feature alignment and fusion, unifying the resolution of each scale segmentation mask, calculating a cross-scale similarity coefficient to identify a divergence region, and generating a fused segmentation result containing a cytoplasm mask, a cell nucleus mask and a complete cell region. Therefore, through multi-resolution scale image acquisition and differentiation processing, in combination with cross-scale feature alignment, dynamic weight fusion and bifurcation region identification, the problems that a single-resolution image is difficult to consider global and details in existing cell fusion degree analysis, a traditional segmentation method is insufficient in precision, and multi-scale feature fusion is low in reliability are effectively solved.

Disclosed are methods for isolating an analyte from a sample. In some aspects, the methods are for selectively isolating a microbial cell analyte, such as a nucleic acid, from a sample containing or suspected of containing mammalian cells. The selective isolation method includes selective lysis of the mammalian cells and filtration of the resulting lysate through a filter that retains intact microbial cells, followed by on-filter lysis of the retained microbial cells to release the microbial cell analyte. The released analyte is then eluted from the filter. In other aspects, the methods utilize on-filter lysis of a sample containing intact cells (e.g., microbial cells) to release the analytes, followed by elution of the released analytes from the filter by passing an immiscible liquid through the filter. The isolated analytes may be analyzed using a suitable assay depending on the type of analyte molecule. Also disclosed are fluidic systems and lysis solutions that may be used for isolating an analyte according to the disclosed methods.

The invention discloses a tissue cell nucleus extraction method and a cell nucleus protective agent applied to flow sorting, and relates to the technical field of cell extraction.The tissue cell nucleus extraction method comprises the steps that operation instruments needed by an experiment are prepared and pre-cooled; cutting a fresh tissue sample into tissue blocks, and immediately transferring the tissue blocks into a precooled lysis buffer solution; manually grinding the lysis buffer solution containing the tissue block by using a homogenizer, and sampling and observing homogenate in the grinding process; filtering the homogenate product through a cell filter screen in sequence; performing low-speed centrifugation on the filtrate by adopting a centrifugal machine, and discarding supernatant to obtain primary cell nucleus precipitate; re-suspending the primary cell nucleus precipitate by using a cell nucleus washing buffer solution to obtain a purified cell nucleus precipitate; and precipitating and resuspending the purified cell nucleuses, counting by trypan blue dyeing, and calculating the percentage of the complete cell nucleuses. According to the method, the problem of more cell nucleus breakage in the tissue cell nucleus extraction process is solved through a mild homogenization strategy monitored under the microscope.

The invention relates to a method for enriching a cell population containing intact and damaged cells in a liquid medium, comprising the steps of: a) providing the liquid medium containing the cell population, b) sedimenting the cell population from the liquid medium, c) separating the liquid medium and the sedimented cell population, d) resuspending the sedimented cell population in a suspension solution to produce a cell suspension, e) adding a ferromagnetic material to the cell suspension that binds specifically to the damaged cells, f) incubating the cell suspension containing the ferromagnetic material, g) placing the cell suspension in a magnetic field to separate the bound and unbound cells in the cell suspension, and h) separating a portion of the cell suspension enriched with unbound, intact cells.the use of intact sperm cells enriched in the process and a device for separating cells bound to magnetic particles in a cell suspension.

In one aspect, the disclosure relates to a process for purification of hydrophobic proteins and / or proteins that have a tendency to aggregate when produced at high levels in cell culture. In a further aspect, the process involves subjecting intact cells to a strong base in a solution with a pH of greater than 12.5, with an optional initial pretreatment at a pH of greater than 13. The process operates efficiently with a high biomass loading of up to or greater than about 20% (w / v) and does not require the use of external flocculants. The target protein can purified by the disclosed process inexpensively and efficiently, producing a high-quality end product having few to no contaminants. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

The present invention relates to the field of biotechnology and a method for creating a cDNA library. More specifically, the invention refers to a method of forming complementary DNA (cDNA) sequencing libraries from RNA in situ comprising the steps of: (a) fixating cells, immobilized on a solid surface; (b) performing an in situ reverse transcription (RT) inside cells, using RT primers comprising a PCR handle 1, and partially biotinylated dNTPs; (c) releasing single stranded cDNA from the cells using a release mix, wherein the release mix comprises an RNAse, such that the single stranded cDNA is released from intact cells; (d) collecting a supernatant comprising released cDNA into a single larger volume or in separate volumes; and (e) introducing an adapter molecule comprising a PCR handle 2 that will bind 3′ of the extended cDNA.

A surgical treatment apparatus comprises a waterjet configured to fragment tissue and provide intact cells such as stem cells with the fragmented tissue. The intact cells can be used in one or more of many ways such as for genetic or other testing. In many embodiments, the intact cells comprise stem cells. In many embodiments, a waterjet is configured to fragment tissue. The fragmented tissue can be collected with a filter having pores sized smaller than the tissue fragments. In many embodiments cavitation with a waterjet is used to fragment the tissue comprising the intact stem cells. The waterjet may comprise a waterjet immersed in a liquid comprising water so as to form a plurality of shedding pulses. The plurality of shedding pulses can be generated with a frequency sufficient to fragment the tissue. The shedding pulses can generate cavitations that fragment the tissue.

The invention belongs to the technical field of biological medicine and clinical medicine, and particularly relates to an efficient purification method of brain tissue small extracellular vesicles. The purification method comprises the following steps: cutting mouse brain tissues into uniform small blocks by using a steel wire mesh, digesting and dissociating, filtering, taking filtrate, and fixing the volume of the filtrate by using an EBSS buffer solution to obtain a tissue supernatant; the method comprises the following steps: sequentially removing complete cells, cell debris, plasma membranes and large vesicles through gradient centrifugation, filtering, taking filtrate, carrying out ultracentrifugation to obtain sEVs precipitates, and resuspending the sEVs precipitates with a PBS buffer solution to obtain a sEVs crude purification product; and preparing a single sucrose pad from a sucrose solution with the concentration of 25-35% and a PBS buffer solution, and carrying out ultracentrifugation on the sEVs crude purification product, wherein the obtained precipitate is the purified brain tissue sEVs. The purification method provided by the invention solves the problems of tedious flow, long consumed time, low yield, poor purity and the like in the existing sEVs purification technology.

The present invention relates to the technical field of cell classification, and particularly relates to a fully automatic classification detection method for peripheral blood cell morphology. The method acquires cell regions in a cell display image of peripheral blood; obtains overlapping cell regions according to the morphology of the cell regions and the curvature of edge pixel points in the cell regions; obtains the exposed edge lines of the covered cells according to the length of each edge line in the overlapping cell regions, the curvature of the edge pixel points on each edge line, and the curvature of the intersection pixel points of each edge line with other edge lines; determines the complete cell edge lines of the covered cells according to the curvature and endpoints of the exposed edge lines, and performs fully automatic classification detection of peripheral blood cell morphology. By acquiring the complete cell edge lines of the covered cells, the present invention determines the morphology of the covered cells, avoids the situation of inaccurate automatic recognition and classification of cells due to cell overlap, and improves the accuracy of fully automatic classification detection of peripheral blood cell morphology.

A surgical treatment apparatus comprises a waterjet configured to fragment tissue and provide intact cells such as stem cells with the fragmented tissue. The intact cells can be used in one or more of many ways such as for genetic or other testing, and the intact cells can be identified as stem cells. In many embodiments, the intact cells comprise stem cells. In many embodiments, a waterjet is configured to fragment tissue. The fragmented tissue can be collected with a filter having pores sized smaller than the tissue fragments. In many embodiments cavitation with a waterjet is used to fragment the tissue comprising the intact stem cells. The waterjet may comprise a waterjet immersed in a liquid comprising water so as to form a plurality of shedding pulses. The plurality of shedding pulses can be generated with a frequency sufficient to fragment the tissue. The shedding pulses can generate cavitations that fragment the tissue.

The present application relates to the technical field of plant-derived nanovesicle, and discloses a preparation method, preparation and composition of plant extracellular vesicles derived from phellodendri cortex. The fruit, pericarp, pulp or leaf of phellodendri cortex is used as raw material to extract liquid under the condition of 0-30 DEG C by pressing, crushing or homogenization. The phellodendri cortex plant material can be treated by microjet high pressure homogenization, and the temperature rise is controlled. At least two levels of clarification treatment are used to remove intact cells, fibers, tissue fragments and large particle impurities, wherein at least one level is filtration treatment, and centrifugal separation is an optional step. Then, the liquid is concentrated and replaced by ultrafiltration or tangential flow filtration, and size exclusion chromatography purification is performed, and the membrane nanoparticle enriched fraction is collected. The obtained particles can be characterized by transmission electron microscopy, nanoparticle tracking analysis and / or dynamic light scattering in terms of morphology, particle size distribution and membrane properties, and the quality control indicators such as candidate particle size range, particle concentration and membrane integrity are established according to the detection results. The particles can include plant extracellular vesicles derived from phellodendri cortex and / or membrane nanovesicles derived from phellodendri cortex, and the obtained preparation can be prepared into a composition, a freeze-dried powder or an oral preparation.

The present invention belongs to the field of biotechnology, and specifically relates to a method for separating bacterial membrane proteins, comprising the following steps: using an osmotic shock method to separate periplasmic proteins from bacterial liquid, resuspending and incubating the remaining precipitate, and then ultrasonically crushing it to obtain an ultrasonic lysate; removing intact cells from the ultrasonic lysate and centrifuging it, the resulting supernatant is cytoplasmic components and outer membrane components, and the precipitate is resuspended; adding anionic surfactants to the resuspended precipitate, incubating, centrifuging, the resulting supernatant is inner membrane protein, and the precipitate is resuspended to be outer membrane protein. The separation method provided by the present invention is not limited by high-end instruments, and can simply, efficiently and quickly separate protein components of bacterial cells at all levels.

A method of producing cell derived particles is disclosed. The method comprising isolating cell-derived particles from a biological sample comprising cells modified to present CD24 so as to obtain a preparation of the cell-derived particles substantially devoid of intact cells. Cell derived particles, a culture medium and a cell culture are also disclosed.

The invention discloses a single cell printing equipment control method and system based on convolutional neural network optimization, and relates to the technical field of intelligent control, and the control method comprises the steps: continuously collecting cell suspension images containing a target single cell before jet printing operation is carried out on a cell suspension, so as to determine each frame of cell main region image, after inputting the shallow convolutional network, respectively constructing a pixel gray parameter set, a pixel response parameter set and a pixel mutation parameter set; analyzing the morphological fluctuation degree of each frame of cell main region image to screen out a standard morphological feature image; the integrity degree of a target single cell structure contained in the standard morphological feature image is analyzed, a cell integrity index is determined to judge whether the integrity degree of the target single cell structure in the current standard morphological feature image meets the printing requirement or not, and a printing control instruction of a corresponding grade is sent out and executed; accurate identification of complete cells in the cell suspension is achieved, and the printing yield and stability are improved.

A method of producing cell derived particles is disclosed. The method comprising isolating cell-derived particles from a biological sample comprising cells modified to present CD24 so as to obtain a preparation of the cell-derived particles substantially devoid of intact cells. Cell derived particles, a culture medium and a cell culture are also disclosed.

The utility model relates to a detachable and combinable cell membrane flow inlay dynamic model, which comprises a plurality of arc-shaped substrates which are detachably connected end to end to form an annular structure; the first aluminum strips are arranged on the arc-shaped base plates in the circumferential direction of the arc-shaped base plates at equal intervals and are close to the inner side edges of the arc-shaped base plates; and the second aluminum strips are arranged on the arc-shaped substrates at equal intervals in the circumferential direction of the arc-shaped substrates and are close to the outer side edges. According to the detachable and combinable cell membrane flow inlay dynamic model designed by the utility model, through the plurality of detachable and combinable arc-shaped substrates, components of a cell membrane can be intuitively and three-dimensionally displayed, and each arc-shaped substrate unit can be independently assembled, so that group teaching and student practice operation are facilitated; meanwhile, all the arc-shaped substrate units can be quickly and conveniently spliced and combined into a complete cell membrane model of an annular structure, overall display can be conveniently carried out on magnetic surfaces such as a blackboard in a classroom, and the overall structure of the cell membrane is visually presented.

The present invention relates to an apparatus for disrupting cell walls-free cells without disrupting a biologically active compartment contained therein surrounded by lipid bilayers and a method of using said apparatus, wherein shear force generating means (S) generate a shear force region suitable for disrupting said cells. Optionally, a pump (P) for pumping an aqueous medium containing the cells at an adjustable flow rate or flow rate, preferably separately adjusting residence time and shear force, and a separating device (C) for separating the at least one released bioactive compartment and / or the bioactive liquid phase around it from the debris are combined. The equipment is an integral part of a stand-alone system or production line. The apparatus and methods are capable of producing products comprising high yields of bioactive compartments that exhibit high biological activity, particularly in energy regeneration and / or protein synthesis.

The application discloses a single-cell printing equipment control method and system based on convolutional neural network optimization, and relates to the technical field of intelligent control. The control method comprises the following steps: before performing a jet printing operation on a cell suspension, continuously collecting cell suspension images containing target single cells to determine cell main area images, and after inputting a shallow convolutional network, respectively constructing a pixel gray parameter set, a pixel response parameter set and a pixel mutation parameter set; analyzing the morphological fluctuation degree of each cell main area image to screen out a standard morphological feature image; analyzing the structural integrity degree of the target single cells contained in the standard morphological feature image to determine a cell integrity index, so as to judge whether the structural integrity degree of the target single cells in the current standard morphological feature image meets the printing requirements, and to issue corresponding level printing control instructions and perform the printing control instructions. The application realizes accurate identification of complete cells in the cell suspension, and improves the yield rate and stability of printing.

The present disclosure is directed to a non-enzymatic dissociation method which facilitates the dissociation of one or more formalin-fixed paraffin-embedded tissue samples into dissociated single cells. The present disclosure is also directed to methods of single cell analysis, i.e., methods of analyzing and / or measuring target components (e.g., biomolecules such as, but not limited to, polypeptides, polynucleotides, small molecules, and the like) on or in cells non-enzymatically dissociated from one or more FFPE tissue samples.

The present invention relates to the technical field of cell membrane permeability detection, and specifically to a method for quantitative detection of cell membrane permeability Δ conductivity. By studying the cell membrane permeability of the first cotyledon and the second cotyledon in the two-leaf-one-core stage of wheat, it was found that for cells with intact cell membranes, the amount of electrolyte transport across the membrane is linear within a specific time period. By detecting and calculating the changes in the conductivity of the exudate during this period through various parameters, the cell membrane permeability can be accurately reflected, and the repeatability and reliability of the results can meet the standards of quantitative research, that is, quantitative detection of cell membrane permeability is achieved; it is verified and summarized that the Δ conductivity method is used to detect cell membrane permeability, which is particularly suitable for qualitative and quantitative detection of plant cell membrane permeability. At the same time, the Δ conductivity method has application prospects in the quantitative detection of cell membrane permeability in large fungi and bacterial microorganisms. The present application provides a special conductivity detector to achieve automatic detection, avoid manual operation errors, and have high detection efficiency.