88 results about "Nucleated Red Blood Cell" patented technology

The present invention provides systems, apparatuses, and methods to isolate, select or detect the presence of a target cell (e.g., fetal cells) in a sample comprising mixed populations of cells that vastly outnumber the target cells. Target cells include fetal cells, such as nucleated red blood cells, and methods of selecting such cells include diagnosis of fetal abnormalities, i.e., aneuploidy. Furthermore, methods comprise utilizing fetal biomarkers to select fetal cells in a sample comprising fetal and adult cells.

The invention relates to a method of identifying fetal abnormality from a maternal blood sample by capturing an image of a fetal nucleated red blood cell obtained from the maternal blood sample; inputting probe intensities for a plurality of nucleic acid probes that bind fetal nucleic acids of interest; analyzing the probe intensities; and generating a diagnostic output according to results of the analysis. In some embodiments, the probes are specific to a chromosome.

A method for enumerating white blood cells and nucleated red blood cells. The method comprises the steps of:(b) providing a lysed sample of whole blood;(b) introducing the lysed sample to a light-scattering multi-angle depolarizing flow cytometer;(c) removing depolarizing interference, e.g., lipid droplets and other measured particles;(d) differentiating nucleated red blood cells and noise from white blood cells in the absence of depolarizing interference;(e) differentiating nucleated red blood cells from noise in the absence of depolarizing interference and white blood cells; and(f) differentiating possible platelet clumps.

Methods are provided for measurement of nucleated red blood cells. The methods include exposing a blood cell sample to a reagent system to lyse mature red blood cells, subsequently analyzing nucleated red blood cells in a flow cell by axial light loss, low angle light scatter and DC impedance measurements, or two selected measurements thereof; and then differentiating nucleated red blood cells from other cell types by using measured signals and / or functions thereof.

Methods are provided for measurement of nucleated red blood cells in a blood sample. The methods include exposing a blood cell sample to a reagent system to lyse mature red blood cells, subsequently analyzing nucleated red blood cells in a flow cell by axial light loss, DC impedance, and medium angle light scatter measurements; by axial light loss, low angle light scatter, and medium angle light scatter measurements; or by axial light loss, DC impedance, low angle light scatter, and medium angle light scatter measurements; and then differentiating nucleated red blood cells from other cell types by using measured signals and / or functions thereof.

A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes. Populations of hematological cells are detected and enumerated using at least two parameters (fluorescence, optical, and electrical) for each population.

Methods are provided for measurement of nucleated red blood cells in a blood sample. The methods include exposing a blood cell sample to a reagent system to lyse mature red blood cells, subsequently analyzing nucleated red blood cells in a flow cell by axial light loss, DC impedance, and medium angle light scatter measurements; by axial light loss, low angle light scatter, and medium angle light scatter measurements; or by axial light loss, DC impedance, low angle light scatter, and medium angle light scatter measurements; and then differentiating nucleated red blood cells from other cell types by using measured signals and / or functions thereof.

A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes. Populations of hematological cells are detected and enumerated using at least two parameters (fluorescence, optical, and electrical) for each population.

The present invention is drawn to a hematology control made from particles a particle having a biopolymer attached to a surface of the particle. The particle simulates a component of a blood sample, such as a reticulocyte or nucleated red blood cell component of a blood cell sample in a flow cytometer or hematology analysis instrument. The present invention is further drawn to methods of making and using the hematology control.

A method of making a reference control containing a nucleated red blood cell component includes providing a blood cell containing a nucleus; treating the blood cell with a treatment solution to alter a nucleus property from a natural value to a target value suitable for simulating nucleated red blood cells on a blood analyzer; and suspending treated blood cell in a suspension medium to form a reference control. The method also includes integrating the nucleated red blood cell component with white blood cell, red blood cell, platelet and reticulocyte components. Further disclosed is a cell treatment composition for altering a nucleus property, which includes a conditioning component, a lytic component for permeating cell membrane, and a fixing component for preserving the cell nucleus. Also disclosed is a method of using the reference control for measurement of nucleated red blood cells on a blood analyzer.

Methods of using a reference control composition containing a nucleated red blood cell component for measurement of nucleated red blood cells are disclosed. The nucleated red blood cell component is made of stabilized or processed nucleated blood cells which have impedance and optical properties simulating the impedance and optical properties of human nucleated red blood cells under a blood lysing condition as measured by a specific measurement. The methods include mixing the reference control composition with a lytic reagent, and analyzing the control sample mixture on a flow cytometric instrument by impedance, impedance and optical measurement, or DC impedance and a multi-dimensional measurement of light scatter, radio frequency and a second DC impedance; and reporting the simulated nucleated red blood cells in the reference control composition.

A reagent for measuring a basophil and / or a nucleated red blood cell is described. The reagent comprises (a) an alcohol having 4 to 8 total carbon atoms, and (b) one or more fluorescent dyes selected from the group consisting of a compound of the general formula (I) and a compound of the general formula (II).

A method of making a reference control containing a nucleated red blood cell component includes providing a blood cell containing a nucleus; treating the blood cell with a treatment solution to alter a nucleus property from a natural value to a target value suitable for simulating nucleated red blood cells on a blood analyzer; and suspending treated blood cell in a suspension medium to form a reference control. The method also includes integrating the nucleated red blood cell component with white blood cell, red blood cell, platelet and reticulocyte components. Further disclosed is a cell treatment composition for altering a nucleus property, which includes a conditioning component, a lytic component for permeating cell membrane, and a fixing component for preserving the cell nucleus. Also disclosed is a method of using the reference control for measurement of nucleated red blood cells on a blood analyzer.

Reference control compositions and the method of use are disclosed for measurement of nucleated red blood cells, which includes one set of synthetic spherical particles having a mean particle diameter ranging from 6.2 μm to 6.8 μm and a refractive index from 1.58 to 1.62 monodispersed in an aqueous suspension medium. The synthetic spherical particles have optical properties simulating optical properties of nucleated red blood cells as measured by optical measurements. The reference control composition can further include a second set of the synthetic spherical particles having optical properties simulating optical properties of white blood cells. Further disclosed is a reference control system which includes a series of reference control compositions, each having an amount of one type of synthetic spherical particles which have optical properties simulating the optical properties of nucleated red blood cells having a specific cell maturity.

Methods are provided for measurement of nucleated red blood cells. The methods include exposing a blood cell sample to a reagent system to lyse mature red blood cells, subsequently analyzing nucleated red blood cells in a flow cell by axial light loss, low angle light scatter and DC impedance measurements, or two selected measurements thereof; and then differentiating nucleated red blood cells from other cell types by using measured signals and / or functions thereof.

The invention discloses a method for separating and appraising nucleated red blood cells in blood, including the following steps: (1) confecting reagent; (2) preparing anti-degumming slide; (3) selecting the staining method for nucleated red blood cells; (4) getting nucleated red blood cells from blood through microoperation. The method for separating and appraising nucleated red blood cells in blood can effectively identify nucleated red blood cells. Nucleated blood cell can be detected in 40 samples and the detection rate is 100 percent. Each sample is detected to have 2*106 to 6*106 / ml nucleated blood cell. In the invention, after the aniline being stained, the nucleated red blood cell is easy to be identified and skilled microoperation can quickly obtain the single nucleated red blood cell, so that the invention lays a good primary foundation to apply the single red blood cell in gene analysis.

A method for measuring immature granulocytes in a blood sample is described. The method includes the steps of lysing red blood cells of a blood sample with a lytic reagent, analyzing the sample mixture by DC impedance measurement, determining immature granulocytes from obtained DC histogram, and reporting immature granulocytes in the blood sample. The method further includes measuring nucleated red blood cells in the blood sample.

A method for concentrating and isolating nucleated cells, such as maternal and fetal nucleated red blood cells (nRBCs), in a maternal whole blood sample. The invention also provides methods and apparatus for preparing to analyze and analyzing the sample for identification of fetal genetic material as part of prenatal genetic testing. The invention also pertains to methods and apparatus for discriminating fetal nucleated red blood cells from maternal nucleated red blood cells obtained from a blood sample taken from a pregnant woman.

The present invention is drawn to a hematology control made from particles a particle having a biopolymer attached to a surface of the particle. The particle simulates a component of a blood sample, such as a reticulocyte or nucleated red blood cell component of a blood cell sample in a flow cytometer or hematology analysis instrument. The present invention is further drawn to methods of making and using the hematology control.

Systems and methods for analyzing blood samples, and more specifically for performing a nucleated red blood cell (nRBC) analysis. The systems and methods screen a blood sample by means of fluorescence staining and a fluorescence triggering strategy, to identify nuclei-containing particles within the blood sample. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder reagent(s), suitable for assays of samples containing fragile white blood cells (WBCs). In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; (b) using a fluorescence trigger to screen the blood sample for nuclei-containing particles; and (c) using measurements of light scatter and fluorescence emission to distinguish nRBCs from WBCs.

A method to determine an engrafting cell dose of hematopoietic stem cell transplant units from transplant sources having nucleated cells selected from the group consisting of cord blood, bone marrow, peripheral blood comprising the steps of[0002]subjecting the source to a substantially complete erythrocyte lysis,[0003]measuring in a cell counter a signal corresponding selectively to white blood cells,[0004]assessing essentially quantitatively nucleated red blood cell (NRBC) count as part of the total nucleated cell (NC) count,[0005]and determining the number of white blood cells (WBCs) as transplant relevant cells.

The invention discloses a separation and purification method for fetal nucleated red blood cells and a Down syndrome screening kit and belongs to the detection field of molecular cell biology. The separation and purification method comprises the following steps: firstly, fully mixing maternal peripheral blood and a CD45 antibody labeling magnetic bead to adsorb white blood cells; discarding on a magnetic separator, transferring upper-layer cells to a test tube containing a CD71 antibody labeling magnetic bead, and fully and uniformly mixing; separating on the magnetic separator to obtain nucleated red blood cells; extracting a genome DNA (Deoxyribonucleic Acid) of the nucleated red blood cells; adding SRY (Sexdetermining Region Y), DSCR (Down Syndrome Critical Region) and USC2 specific probes, DSCR and USC2 competitive primers, an SRY specific primer and a DNA to be detected into the same competitive fluorescent quantitation PCR (Polymerase Chain Reaction) reaction system for multiple competitive real-time fluorescence quantification PCR reactions to obtain Cts of DSCR and USC2; and calculating a difference value between the Cts to judge whether a sample to be detected is highly risk to DS (Down Syndrome).

The invention provides a blood cell separation system, which is used for the precise separation and concentration of rare fetal nucleated cells mixed in the blood of pregnant women, so as to facilitate the obtaining of test preparations that can be used for prenatal chromosome / gene diagnosis. The blood cell separation system is characterized in that it includes (1) a primary separation device, which is used to mainly remove non-nucleated red blood cells, white blood cells and platelets from a blood sample taken from a pregnant woman to obtain a primary separation sample, (2) a secondary separation device , for the removal of residual non-nucleated erythrocytes and leukocytes from a primary fraction obtained using the primary fractionation device using the carbohydrate-lectin method to obtain a secondary fraction containing concentrated fetal nucleated cells, and (3) A preparation device for preparing the secondary separation sample obtained by the secondary separation device.

Methods of using a reference control composition containing a nucleated red blood cell component for measurement of nucleated red blood cells are disclosed. The nucleated red blood cell component is made of stabilized or processed nucleated blood cells which have impedance and optical properties simulating the impedance and optical properties of human nucleated red blood cells under a blood lysing condition as measured by a specific measurement. The methods include mixing the reference control composition with a lytic reagent, and analyzing the control sample mixture on a flow cytometric instrument by impedance, impedance and optical measurement, or DC impedance and a multi-dimensional measurement of light scatter, radio frequency and a second DC impedance; and reporting the simulated nucleated red blood cells in the reference control composition.

The invention provides a nucleated red blood cell simulation particle and a preparation method thereof. The particle is a white blood cell combined with a fluorescent dyeing inhibitor, and the inhibitor is stably combined with a nucleus or nucleic acid in the cell, so that the capacity of the particle of combining with a fluorescent dye during detection is reduced. The method comprises the following steps of: (a) obtaining the washed and purified white blood cell; (b) suspending the white blood cell in fluorescent dyeing inhibitor-containing cell processing liquid, wherein the inhibitor is stably combined with the nucleus or the nucleic acid in the cell; and (c) washing the obtained product. The invention also provides a nucleated red blood cell simulation particle-containing blood quality control substance, and application of the nucleated red blood cell simulation particle and the blood quality control substance thereof to the quality control of a blood cell analyzer.

A method for concentrating and isolating nucleated cells, such as maternal and fetal nucleated red blood cells (nRBCs), in a maternal whole blood sample. The invention also provides methods and apparatus for preparing to analyze and analyzing the sample for identification of fetal genetic material as part of prenatal genetic testing. The invention also pertains to methods and apparatus for discriminating fetal nucleated red blood cells from maternal nucleated red blood cells obtained from a blood sample taken from a pregnant woman.

The invention provides a method for capturing and identifying fetal nucleated red blood cells in a microfluidic chip. The method comprises the following steps: 1, synthesizing gelatin nanoparticles bya two-step solvent removal method and preparing into a precursor liquid, and then introducing into a microfluidic chip to form a nanometer coating on the inner wall thereof, wherein the microfluidicchip comprises a plurality of serpentine main channels and the plurality of fish bone units; 2, modifying streptavidin on the nanometer coating by a chemical modification method, then grafting a biotin-modified antibody; 3, separating a mononuclear cell layer containing fetal nucleated red blood cells from peripheral blood by density-gradient centrifugation; 4, allowing mononuclear cell suspensionto flow into the microfluidic chip at a certain flow rate for capturing; 5, introducing a certain concentration of gelatinase into the microfluidic chip that captures fetal nucleated red blood cells,releasing fetal nucleated red blood cells by degrading the nanometer coating; and 6, identifying captured fetal nucleated red blood cells by using specific antibodies.

A linearity control system includes a series of linearity control compositions, each thereof includes white blood cell analogs and stabilized red blood cells in a suspension medium. The concentration of the white blood cell analogs in the series of control compositions increases from 0.2×103 to 800×103 analogs per microliter, and the concentration of the white blood cell analogs in at least one control composition is greater than 120×103 analogs per microliter. The stabilized red blood cells facilitate mono-dispersion of the white blood cell analogs in the suspension medium by gently mixing. The control compositions further include platelet analogs, or additionally include reticulocyte and / or nucleated red blood cell analogs. The linearity control system allows the verification of the reportable measurement range and linearity of the measurements of hematology analyzers for white blood cells, red blood cells and platelets in extended concentration ranges.

This invention provides a method for concentrating and collecting small quantities of fetal nucleated red blood cells contained in the maternal blood. The method for concentrating and collecting nucleated red blood cells from the maternal blood comprises: (i) subjecting the maternal blood to a first density-gradient centrifugation and collecting a cell fraction containing nucleated red blood cells; (ii) treating the cell fraction containing nucleated red blood cells so as to selectively changes the density of the nucleated red blood cells from that of the white blood cells; and (iii) subjecting the treated cell fraction containing the nucleated red blood cells to a second density-gradient centrifugation so as to collect a fraction containing nucleated red blood cells.

The present invention relates to a method for separating free fetal cells from peripheral blood of a pregnant woman. The method utilizes the cell separation function and the specific affinity recognition principle of a microfluidic chip to realize high-efficiency and high-purity capture and separation of the free fetal cells (fetal nucleated red blood cells, trophoblast cells, and the like) in theperipheral blood of the pregnant woman. Within the microfluidic chip, a microarray structure with a specific geometric arrangement is designed, and the microarray structure is modified with moleculargroups having an affinity recognition function such as a nucleic acid aptamer, an antibody and a polypeptide. By adjusting parameters of the microarray structure, the collision efficiency of different sizes of fetal cells and microarrays can be controlled, and the capture and enrichment of the fetal cells of different sizes can be realized.